Publications
Browse peer-reviewed literature, posters, webinars, blog articles, and more showing how we and others are using RepliGut Systems to support discovery.
2024
Pike, Colleen M.; Levi, James A.; Boone, Lauren A.; Peddibhotla, Swetha; Johnson, Jacob; Zwarycz, Bailey; Bunger, Maureen K.; Thelin, William; Boazak, Elizabeth M.
High-Throughput Assay for Predicting Diarrhea Risk Using a 2D Human Intestinal Stem Cell-Derived Model Miscellaneous
2024, (Pages: 2024.08.28.610072 Section: New Results).
Abstract | Links | BibTeX | Tags:
@misc{pike_high-throughput_2024-1,
title = {High-Throughput Assay for Predicting Diarrhea Risk Using a 2D Human Intestinal Stem Cell-Derived Model},
author = {Colleen M. Pike and James A. Levi and Lauren A. Boone and Swetha Peddibhotla and Jacob Johnson and Bailey Zwarycz and Maureen K. Bunger and William Thelin and Elizabeth M. Boazak},
url = {https://www.biorxiv.org/content/10.1101/2024.08.28.610072v1},
doi = {10.1101/2024.08.28.610072},
year = {2024},
date = {2024-08-28},
urldate = {2025-03-11},
publisher = {bioRxiv},
abstract = {Gastrointestinal toxicities (GITs) are the most prevalent adverse events (AE) reported in clinical trials, often resulting in dose-limitations that reduce drug efficacy and delay development and treatment optimization. Preclinical animal models do not accurately replicate human GI physiology, leaving few options for early detection of GI side effects prior to human studies. Development of an accurate model that predicts GIT earlier in drug discovery programs would better support successful clinical trial outcomes. Chemotherapeutics, which exhibit high rates of clinical GIT, frequently target mitotic cells. Therefore, we hypothesized that a model utilizing proliferative cell populations derived from human intestinal crypts would predict the occurrence of clinical GITs with high accuracy. Here, we describe the development of a multiparametric assay utilizing the RepliGut® Planar system, an intestinal stem cell-derived platform cultured in an accessible high throughput Transwell™ format. This assay addresses key physiological elements of GIT by assessing cell proliferation (EdU incorporation), cell abundance (DAPI quantification), and barrier function (TEER). Using this approach, we demonstrate that primary proliferative cell populations reproducibly respond to marketed chemotherapeutics at physiologic concentrations. To determine the ability of this model to predict clinical diarrhea risk, we evaluated a set of 30 drugs with known clinical diarrhea incidence in three human donors, comparing results to known plasma drug concentrations. This resulted in highly accurate predictions of diarrhea potential for each endpoint (balanced accuracy of 91% for DAPI, 90% for EdU, 88% for TEER) with minimal variation across human donors. In vitro toxicity screening using primary proliferative cells may enable improved safety evaluations, reducing the risk of AEs in clinical trials and ultimately lead to safer and more effective treatments for patients.},
note = {Pages: 2024.08.28.610072
Section: New Results},
keywords = {},
pubstate = {published},
tppubtype = {misc}
}
Gastrointestinal toxicities (GITs) are the most prevalent adverse events (AE) reported in clinical trials, often resulting in dose-limitations that reduce drug efficacy and delay development and treatment optimization. Preclinical animal models do not accurately replicate human GI physiology, leaving few options for early detection of GI side effects prior to human studies. Development of an accurate model that predicts GIT earlier in drug discovery programs would better support successful clinical trial outcomes. Chemotherapeutics, which exhibit high rates of clinical GIT, frequently target mitotic cells. Therefore, we hypothesized that a model utilizing proliferative cell populations derived from human intestinal crypts would predict the occurrence of clinical GITs with high accuracy. Here, we describe the development of a multiparametric assay utilizing the RepliGut® Planar system, an intestinal stem cell-derived platform cultured in an accessible high throughput Transwell™ format. This assay addresses key physiological elements of GIT by assessing cell proliferation (EdU incorporation), cell abundance (DAPI quantification), and barrier function (TEER). Using this approach, we demonstrate that primary proliferative cell populations reproducibly respond to marketed chemotherapeutics at physiologic concentrations. To determine the ability of this model to predict clinical diarrhea risk, we evaluated a set of 30 drugs with known clinical diarrhea incidence in three human donors, comparing results to known plasma drug concentrations. This resulted in highly accurate predictions of diarrhea potential for each endpoint (balanced accuracy of 91% for DAPI, 90% for EdU, 88% for TEER) with minimal variation across human donors. In vitro toxicity screening using primary proliferative cells may enable improved safety evaluations, reducing the risk of AEs in clinical trials and ultimately lead to safer and more effective treatments for patients.
Pike, Colleen M.; Levi, James A.; Boone, Lauren A.; Peddibhotla, Swetha; Johnson, Jacob; Zwarycz, Bailey; Bunger, Maureen K.; Thelin, William; Boazak, Elizabeth M.
High-Throughput Assay for Predicting Diarrhea Risk Using a 2D Human Intestinal Stem Cell-Derived Model Miscellaneous
2024, (Pages: 2024.08.28.610072 Section: New Results).
Abstract | Links | BibTeX | Tags:
@misc{pike_high-throughput_2024,
title = {High-Throughput Assay for Predicting Diarrhea Risk Using a 2D Human Intestinal Stem Cell-Derived Model},
author = {Colleen M. Pike and James A. Levi and Lauren A. Boone and Swetha Peddibhotla and Jacob Johnson and Bailey Zwarycz and Maureen K. Bunger and William Thelin and Elizabeth M. Boazak},
url = {https://www.biorxiv.org/content/10.1101/2024.08.28.610072v1},
doi = {10.1101/2024.08.28.610072},
year = {2024},
date = {2024-08-28},
urldate = {2024-11-25},
publisher = {bioRxiv},
abstract = {Gastrointestinal toxicities (GITs) are the most prevalent adverse events (AE) reported in clinical trials, often resulting in dose-limitations that reduce drug efficacy and delay development and treatment optimization. Preclinical animal models do not accurately replicate human GI physiology, leaving few options for early detection of GI side effects prior to human studies. Development of an accurate model that predicts GIT earlier in drug discovery programs would better support successful clinical trial outcomes. Chemotherapeutics, which exhibit high rates of clinical GIT, frequently target mitotic cells. Therefore, we hypothesized that a model utilizing proliferative cell populations derived from human intestinal crypts would predict the occurrence of clinical GITs with high accuracy. Here, we describe the development of a multiparametric assay utilizing the RepliGut® Planar system, an intestinal stem cell-derived platform cultured in an accessible high throughput Transwell™ format. This assay addresses key physiological elements of GIT by assessing cell proliferation (EdU incorporation), cell abundance (DAPI quantification), and barrier function (TEER). Using this approach, we demonstrate that primary proliferative cell populations reproducibly respond to marketed chemotherapeutics at physiologic concentrations. To determine the ability of this model to predict clinical diarrhea risk, we evaluated a set of 30 drugs with known clinical diarrhea incidence in three human donors, comparing results to known plasma drug concentrations. This resulted in highly accurate predictions of diarrhea potential for each endpoint (balanced accuracy of 91% for DAPI, 90% for EdU, 88% for TEER) with minimal variation across human donors. In vitro toxicity screening using primary proliferative cells may enable improved safety evaluations, reducing the risk of AEs in clinical trials and ultimately lead to safer and more effective treatments for patients.},
note = {Pages: 2024.08.28.610072
Section: New Results},
keywords = {},
pubstate = {published},
tppubtype = {misc}
}
Gastrointestinal toxicities (GITs) are the most prevalent adverse events (AE) reported in clinical trials, often resulting in dose-limitations that reduce drug efficacy and delay development and treatment optimization. Preclinical animal models do not accurately replicate human GI physiology, leaving few options for early detection of GI side effects prior to human studies. Development of an accurate model that predicts GIT earlier in drug discovery programs would better support successful clinical trial outcomes. Chemotherapeutics, which exhibit high rates of clinical GIT, frequently target mitotic cells. Therefore, we hypothesized that a model utilizing proliferative cell populations derived from human intestinal crypts would predict the occurrence of clinical GITs with high accuracy. Here, we describe the development of a multiparametric assay utilizing the RepliGut® Planar system, an intestinal stem cell-derived platform cultured in an accessible high throughput Transwell™ format. This assay addresses key physiological elements of GIT by assessing cell proliferation (EdU incorporation), cell abundance (DAPI quantification), and barrier function (TEER). Using this approach, we demonstrate that primary proliferative cell populations reproducibly respond to marketed chemotherapeutics at physiologic concentrations. To determine the ability of this model to predict clinical diarrhea risk, we evaluated a set of 30 drugs with known clinical diarrhea incidence in three human donors, comparing results to known plasma drug concentrations. This resulted in highly accurate predictions of diarrhea potential for each endpoint (balanced accuracy of 91% for DAPI, 90% for EdU, 88% for TEER) with minimal variation across human donors. In vitro toxicity screening using primary proliferative cells may enable improved safety evaluations, reducing the risk of AEs in clinical trials and ultimately lead to safer and more effective treatments for patients.
Pike, Colleen M.; Zwarycz, Bailey; McQueen, Bryan E.; Castillo, Mariana; Barron, Catherine; Morowitz, Jeremy M.; Levi, James A.; Phadke, Dhiral; Balik-Meisner, Michele; Mav, Deepak; Shah, Ruchir; Glasspoole, Danielle L. Cunningham; Laetham, Ron; Thelin, William; Bunger, Maureen K.; Boazak, Elizabeth M.
Characterization and optimization of variability in a human colonic epithelium culture model Journal Article
In: vol. 41, no. 3, pp. 425–438, 2024, ISSN: 1868-8551, (Number: 3).
Abstract | Links | BibTeX | Tags: epithelial cell culture, in vitro models, intestinal barrier, Microphysiological systems, permeability
@article{pike_characterization_2024,
title = {Characterization and optimization of variability in a human colonic epithelium culture model},
author = {Colleen M. Pike and Bailey Zwarycz and Bryan E. McQueen and Mariana Castillo and Catherine Barron and Jeremy M. Morowitz and James A. Levi and Dhiral Phadke and Michele Balik-Meisner and Deepak Mav and Ruchir Shah and Danielle L. Cunningham Glasspoole and Ron Laetham and William Thelin and Maureen K. Bunger and Elizabeth M. Boazak},
url = {https://www.altex.org/index.php/altex/article/view/2686},
doi = {10.14573/altex.2309221},
issn = {1868-8551},
year = {2024},
date = {2024-07-16},
urldate = {2025-01-29},
volume = {41},
number = {3},
pages = {425–438},
abstract = {Animal models have historically been poor preclinical predictors of gastrointestinal (GI) directed therapeutic efficacy and drug-induced GI toxicity. Human stem and primary cell-derived culture systems are a major focus of efforts to create biologically relevant models that enhance preclinical predictive value of intestinal efficacy and toxicity. The inherent variability in stem cell-based cultures makes development of useful models a challenge; the stochastic nature of stem cell differentiation interferes with the ability to build and validate reproducible assays that query drug responses and pharmacokinetics. In this study, we aimed to characterize and reduce sources of variability in a complex stem cell-derived intestinal epithelium model, termed RepliGut® Planar, across cells from multiple human donors, cell lots, and passage numbers. Assessment criteria included barrier formation and integrity, gene expression, and cytokine responses. Gene expression and culture metric analyses revealed that controlling cell passage number reduces variability and maximizes physiological relevance of the model. In a case study where passage number was optimized, distinct cytokine responses were observed among four human donors, indicating that biological variability can be detected in cell cultures originating from diverse human sources. These findings highlight key considerations for designing assays that can be applied to additional primary cell-derived systems, as well as establish utility of the RepliGut® Planar platform for robust development of human-predictive drug-response assays.
Plain language summary Animal models are frequently used as tools for studying gastrointestinal (GI) disease, but they inadequately replicate the complexities of the human gut, making them poor predictors of how humans respond to new drugs. Models using human stem cells are closer to human GI physiology, but their responses are not uniform owing to variability in the stem cells. We looked for the sources of this variability in the primary stem-cell derived RepliGut® Planar model. We found that limiting how long the cells were kept in culture reduced their variability and improved the physiological relevance of the model. These findings highlight key assay design considerations that also can be applied to other primary cell-derived systems. Reliable and physiologically relevant cell-based models can reduce animal testing, improve research accuracy, and ensure new treatments are more relevant and effective for patients.},
note = {Number: 3},
keywords = {epithelial cell culture, in vitro models, intestinal barrier, Microphysiological systems, permeability},
pubstate = {published},
tppubtype = {article}
}
Animal models have historically been poor preclinical predictors of gastrointestinal (GI) directed therapeutic efficacy and drug-induced GI toxicity. Human stem and primary cell-derived culture systems are a major focus of efforts to create biologically relevant models that enhance preclinical predictive value of intestinal efficacy and toxicity. The inherent variability in stem cell-based cultures makes development of useful models a challenge; the stochastic nature of stem cell differentiation interferes with the ability to build and validate reproducible assays that query drug responses and pharmacokinetics. In this study, we aimed to characterize and reduce sources of variability in a complex stem cell-derived intestinal epithelium model, termed RepliGut® Planar, across cells from multiple human donors, cell lots, and passage numbers. Assessment criteria included barrier formation and integrity, gene expression, and cytokine responses. Gene expression and culture metric analyses revealed that controlling cell passage number reduces variability and maximizes physiological relevance of the model. In a case study where passage number was optimized, distinct cytokine responses were observed among four human donors, indicating that biological variability can be detected in cell cultures originating from diverse human sources. These findings highlight key considerations for designing assays that can be applied to additional primary cell-derived systems, as well as establish utility of the RepliGut® Planar platform for robust development of human-predictive drug-response assays.
Plain language summary Animal models are frequently used as tools for studying gastrointestinal (GI) disease, but they inadequately replicate the complexities of the human gut, making them poor predictors of how humans respond to new drugs. Models using human stem cells are closer to human GI physiology, but their responses are not uniform owing to variability in the stem cells. We looked for the sources of this variability in the primary stem-cell derived RepliGut® Planar model. We found that limiting how long the cells were kept in culture reduced their variability and improved the physiological relevance of the model. These findings highlight key assay design considerations that also can be applied to other primary cell-derived systems. Reliable and physiologically relevant cell-based models can reduce animal testing, improve research accuracy, and ensure new treatments are more relevant and effective for patients.
Plain language summary Animal models are frequently used as tools for studying gastrointestinal (GI) disease, but they inadequately replicate the complexities of the human gut, making them poor predictors of how humans respond to new drugs. Models using human stem cells are closer to human GI physiology, but their responses are not uniform owing to variability in the stem cells. We looked for the sources of this variability in the primary stem-cell derived RepliGut® Planar model. We found that limiting how long the cells were kept in culture reduced their variability and improved the physiological relevance of the model. These findings highlight key assay design considerations that also can be applied to other primary cell-derived systems. Reliable and physiologically relevant cell-based models can reduce animal testing, improve research accuracy, and ensure new treatments are more relevant and effective for patients.
Corte, Sebastian Gonzalez La; Stevens, Corey A.; Cárcamo-Oyarce, Gerardo; Ribbeck, Katharina; Wingreen, Ned S.; Datta, Sujit S.
Morphogenesis of bacterial colonies in polymeric environments Miscellaneous
2024, (Pages: 2024.04.18.590088 Section: New Results).
Abstract | Links | BibTeX | Tags:
@misc{corte_morphogenesis_2024,
title = {Morphogenesis of bacterial colonies in polymeric environments},
author = {Sebastian Gonzalez La Corte and Corey A. Stevens and Gerardo Cárcamo-Oyarce and Katharina Ribbeck and Ned S. Wingreen and Sujit S. Datta},
url = {https://www.biorxiv.org/content/10.1101/2024.04.18.590088v1},
doi = {10.1101/2024.04.18.590088},
year = {2024},
date = {2024-04-22},
urldate = {2024-11-25},
publisher = {bioRxiv},
abstract = {Many bacteria live in polymeric fluids, such as mucus, environmental polysaccharides, and extracellular polymers in biofilms. However, lab studies typically focus on cells in polymer-free fluids. Here, we show that interactions with polymers shape a fundamental feature of bacterial life—how they proliferate in space in multicellular colonies. Using experiments, we find that when polymer is sufficiently concentrated, cells generically and reversibly form large serpentine “cables” as they proliferate. By combining experiments with biophysical theory and simulations, we demonstrate that this distinctive form of colony morphogenesis arises from an interplay between polymer-induced entropic attraction between neighboring cells and their hindered ability to diffusely separate from each other in a viscous polymer solution. Our work thus reveals a pivotal role of polymers in sculpting proliferating bacterial colonies, with implications for how they interact with hosts and with the natural environment, and uncovers quantitative principles governing colony morphogenesis in such complex environments.},
note = {Pages: 2024.04.18.590088
Section: New Results},
keywords = {},
pubstate = {published},
tppubtype = {misc}
}
Many bacteria live in polymeric fluids, such as mucus, environmental polysaccharides, and extracellular polymers in biofilms. However, lab studies typically focus on cells in polymer-free fluids. Here, we show that interactions with polymers shape a fundamental feature of bacterial life—how they proliferate in space in multicellular colonies. Using experiments, we find that when polymer is sufficiently concentrated, cells generically and reversibly form large serpentine “cables” as they proliferate. By combining experiments with biophysical theory and simulations, we demonstrate that this distinctive form of colony morphogenesis arises from an interplay between polymer-induced entropic attraction between neighboring cells and their hindered ability to diffusely separate from each other in a viscous polymer solution. Our work thus reveals a pivotal role of polymers in sculpting proliferating bacterial colonies, with implications for how they interact with hosts and with the natural environment, and uncovers quantitative principles governing colony morphogenesis in such complex environments.
Sharma, Abhinav; Jin, Liang; Wang, Xue; Wang, Yue-Ting; Stresser, David M.
Developing an adult stem cell derived microphysiological intestinal system for predicting oral prodrug bioconversion and permeability in humans Journal Article
In: vol. 24, no. 2, pp. 339–355, 2024, ISSN: 1473-0189, (Number: 2 Publisher: The Royal Society of Chemistry).
Abstract | Links | BibTeX | Tags:
@article{sharma_developing_2024-1,
title = {Developing an adult stem cell derived microphysiological intestinal system for predicting oral prodrug bioconversion and permeability in humans},
author = {Abhinav Sharma and Liang Jin and Xue Wang and Yue-Ting Wang and David M. Stresser},
url = {https://pubs.rsc.org/en/content/articlelanding/2024/lc/d3lc00843f},
doi = {10.1039/D3LC00843F},
issn = {1473-0189},
year = {2024},
date = {2024-01-17},
urldate = {2024-11-25},
volume = {24},
number = {2},
pages = {339–355},
abstract = {Microphysiological systems (MPS) incorporating human intestinal organoids have shown the potential to faithfully model intestinal biology with the promise to accelerate development of oral prodrugs. We hypothesized that an MPS model incorporating flow, shear stress, and vasculature could provide more reliable measures of prodrug bioconversion and permeability. Following construction of jejunal and duodenal organoid MPS derived from 3 donors, we determined the area under the concentration–time (AUC) curve for the active drug in the vascular channel and characterized the enzymology of prodrug bioconversion. Fosamprenavir underwent phosphatase mediated hydrolysis to amprenavir while dabigatran etexilate (DABE) exhibited proper CES2- and, as anticipated, not CES1-mediated de-esterification, followed by permeation of amprenavir to the vascular channel. When experiments were conducted in the presence of bio-converting enzyme inhibitors (orthovanadate for alkaline phosphatase; bis(p-nitrophenyl)phosphate for carboxylesterase), the AUC of the active drug decreased accordingly in the vascular channel. In addition to functional analysis, the MPS was characterized through imaging and proteomic analysis. Imaging revealed proper expression and localization of epithelial, endothelial, tight junction and catalytic enzyme markers. Global proteomic analysis was used to analyze the MPS model and 3 comparator sources: an organoid-based transwell model (which was also evaluated for function), Matrigel embedded organoids and finally jejunal and duodenal cadaver tissues collected from 3 donors. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) of global proteomic data demonstrated that all organoid-based models exhibited strong similarity and were distinct from tissues. Intestinal organoids in the MPS model exhibited strong similarity to human tissue for key epithelial markers via HCA. Quantitative proteomic analysis showed higher expression of key prodrug converting and drug metabolizing enzymes in MPS-derived organoids compared to tissues, organoids in Matrigel, and organoids on transwells. When comparing organoids from MPS and transwells, expression of intestinal alkaline phosphatase (ALPI), carboxylesterase (CES)2, cytochrome P450 3A4 (CYP3A4) and sucrase isomaltase (SI) was 2.97-, 1.2-, 11.3-, and 27.7-fold higher for duodenum and 7.7-, 4.6-, 18.1-, and 112.2-fold higher for jejunum organoids in MPS, respectively. The MPS approach can provide a more physiological system than enzymes, organoids, and organoids on transwells for pharmacokinetic analysis of prodrugs that account for 10% of all commercial medicines.},
note = {Number: 2
Publisher: The Royal Society of Chemistry},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Microphysiological systems (MPS) incorporating human intestinal organoids have shown the potential to faithfully model intestinal biology with the promise to accelerate development of oral prodrugs. We hypothesized that an MPS model incorporating flow, shear stress, and vasculature could provide more reliable measures of prodrug bioconversion and permeability. Following construction of jejunal and duodenal organoid MPS derived from 3 donors, we determined the area under the concentration–time (AUC) curve for the active drug in the vascular channel and characterized the enzymology of prodrug bioconversion. Fosamprenavir underwent phosphatase mediated hydrolysis to amprenavir while dabigatran etexilate (DABE) exhibited proper CES2- and, as anticipated, not CES1-mediated de-esterification, followed by permeation of amprenavir to the vascular channel. When experiments were conducted in the presence of bio-converting enzyme inhibitors (orthovanadate for alkaline phosphatase; bis(p-nitrophenyl)phosphate for carboxylesterase), the AUC of the active drug decreased accordingly in the vascular channel. In addition to functional analysis, the MPS was characterized through imaging and proteomic analysis. Imaging revealed proper expression and localization of epithelial, endothelial, tight junction and catalytic enzyme markers. Global proteomic analysis was used to analyze the MPS model and 3 comparator sources: an organoid-based transwell model (which was also evaluated for function), Matrigel embedded organoids and finally jejunal and duodenal cadaver tissues collected from 3 donors. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) of global proteomic data demonstrated that all organoid-based models exhibited strong similarity and were distinct from tissues. Intestinal organoids in the MPS model exhibited strong similarity to human tissue for key epithelial markers via HCA. Quantitative proteomic analysis showed higher expression of key prodrug converting and drug metabolizing enzymes in MPS-derived organoids compared to tissues, organoids in Matrigel, and organoids on transwells. When comparing organoids from MPS and transwells, expression of intestinal alkaline phosphatase (ALPI), carboxylesterase (CES)2, cytochrome P450 3A4 (CYP3A4) and sucrase isomaltase (SI) was 2.97-, 1.2-, 11.3-, and 27.7-fold higher for duodenum and 7.7-, 4.6-, 18.1-, and 112.2-fold higher for jejunum organoids in MPS, respectively. The MPS approach can provide a more physiological system than enzymes, organoids, and organoids on transwells for pharmacokinetic analysis of prodrugs that account for 10% of all commercial medicines.
Sharma, Abhinav; Jin, Liang; Wang, Xue; Wang, Yue-Ting; Stresser, David M.
Developing an adult stem cell derived microphysiological intestinal system for predicting oral prodrug bioconversion and permeability in humans Journal Article
In: vol. 24, no. 2, pp. 339–355, 2024, ISSN: 1473-0189, (Publisher: The Royal Society of Chemistry).
Abstract | Links | BibTeX | Tags:
@article{sharma_developing_2024,
title = {Developing an adult stem cell derived microphysiological intestinal system for predicting oral prodrug bioconversion and permeability in humans},
author = {Abhinav Sharma and Liang Jin and Xue Wang and Yue-Ting Wang and David M. Stresser},
url = {https://pubs.rsc.org/en/content/articlelanding/2024/lc/d3lc00843f},
doi = {10.1039/D3LC00843F},
issn = {1473-0189},
year = {2024},
date = {2024-01-17},
urldate = {2024-11-25},
volume = {24},
number = {2},
pages = {339–355},
abstract = {Microphysiological systems (MPS) incorporating human intestinal organoids have shown the potential to faithfully model intestinal biology with the promise to accelerate development of oral prodrugs. We hypothesized that an MPS model incorporating flow, shear stress, and vasculature could provide more reliable measures of prodrug bioconversion and permeability. Following construction of jejunal and duodenal organoid MPS derived from 3 donors, we determined the area under the concentration–time (AUC) curve for the active drug in the vascular channel and characterized the enzymology of prodrug bioconversion. Fosamprenavir underwent phosphatase mediated hydrolysis to amprenavir while dabigatran etexilate (DABE) exhibited proper CES2- and, as anticipated, not CES1-mediated de-esterification, followed by permeation of amprenavir to the vascular channel. When experiments were conducted in the presence of bio-converting enzyme inhibitors (orthovanadate for alkaline phosphatase; bis(p-nitrophenyl)phosphate for carboxylesterase), the AUC of the active drug decreased accordingly in the vascular channel. In addition to functional analysis, the MPS was characterized through imaging and proteomic analysis. Imaging revealed proper expression and localization of epithelial, endothelial, tight junction and catalytic enzyme markers. Global proteomic analysis was used to analyze the MPS model and 3 comparator sources: an organoid-based transwell model (which was also evaluated for function), Matrigel embedded organoids and finally jejunal and duodenal cadaver tissues collected from 3 donors. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) of global proteomic data demonstrated that all organoid-based models exhibited strong similarity and were distinct from tissues. Intestinal organoids in the MPS model exhibited strong similarity to human tissue for key epithelial markers via HCA. Quantitative proteomic analysis showed higher expression of key prodrug converting and drug metabolizing enzymes in MPS-derived organoids compared to tissues, organoids in Matrigel, and organoids on transwells. When comparing organoids from MPS and transwells, expression of intestinal alkaline phosphatase (ALPI), carboxylesterase (CES)2, cytochrome P450 3A4 (CYP3A4) and sucrase isomaltase (SI) was 2.97-, 1.2-, 11.3-, and 27.7-fold higher for duodenum and 7.7-, 4.6-, 18.1-, and 112.2-fold higher for jejunum organoids in MPS, respectively. The MPS approach can provide a more physiological system than enzymes, organoids, and organoids on transwells for pharmacokinetic analysis of prodrugs that account for 10% of all commercial medicines.},
note = {Publisher: The Royal Society of Chemistry},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Microphysiological systems (MPS) incorporating human intestinal organoids have shown the potential to faithfully model intestinal biology with the promise to accelerate development of oral prodrugs. We hypothesized that an MPS model incorporating flow, shear stress, and vasculature could provide more reliable measures of prodrug bioconversion and permeability. Following construction of jejunal and duodenal organoid MPS derived from 3 donors, we determined the area under the concentration–time (AUC) curve for the active drug in the vascular channel and characterized the enzymology of prodrug bioconversion. Fosamprenavir underwent phosphatase mediated hydrolysis to amprenavir while dabigatran etexilate (DABE) exhibited proper CES2- and, as anticipated, not CES1-mediated de-esterification, followed by permeation of amprenavir to the vascular channel. When experiments were conducted in the presence of bio-converting enzyme inhibitors (orthovanadate for alkaline phosphatase; bis(p-nitrophenyl)phosphate for carboxylesterase), the AUC of the active drug decreased accordingly in the vascular channel. In addition to functional analysis, the MPS was characterized through imaging and proteomic analysis. Imaging revealed proper expression and localization of epithelial, endothelial, tight junction and catalytic enzyme markers. Global proteomic analysis was used to analyze the MPS model and 3 comparator sources: an organoid-based transwell model (which was also evaluated for function), Matrigel embedded organoids and finally jejunal and duodenal cadaver tissues collected from 3 donors. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) of global proteomic data demonstrated that all organoid-based models exhibited strong similarity and were distinct from tissues. Intestinal organoids in the MPS model exhibited strong similarity to human tissue for key epithelial markers via HCA. Quantitative proteomic analysis showed higher expression of key prodrug converting and drug metabolizing enzymes in MPS-derived organoids compared to tissues, organoids in Matrigel, and organoids on transwells. When comparing organoids from MPS and transwells, expression of intestinal alkaline phosphatase (ALPI), carboxylesterase (CES)2, cytochrome P450 3A4 (CYP3A4) and sucrase isomaltase (SI) was 2.97-, 1.2-, 11.3-, and 27.7-fold higher for duodenum and 7.7-, 4.6-, 18.1-, and 112.2-fold higher for jejunum organoids in MPS, respectively. The MPS approach can provide a more physiological system than enzymes, organoids, and organoids on transwells for pharmacokinetic analysis of prodrugs that account for 10% of all commercial medicines.
2023
Hu, Qi-rui; Hong, Huan; Zhang, Zhi-hong; Feng, Hua; Luo, Ting; Li, Jing; Deng, Ze-yuan; Chen, Fang
In: vol. 47, no. 6, pp. 694–705, 2023, ISSN: 1226-8453.
Abstract | Links | BibTeX | Tags: ginsenoside, Gut microbiota, oral bioavailability, P-glycoprotein
@article{hu_methods_2023,
title = {Methods on improvements of the poor oral bioavailability of ginsenosides: Pre-processing, structural modification, drug combination, and micro- or nano- delivery system},
author = {Qi-rui Hu and Huan Hong and Zhi-hong Zhang and Hua Feng and Ting Luo and Jing Li and Ze-yuan Deng and Fang Chen},
url = {https://www.sciencedirect.com/science/article/pii/S1226845323000799},
doi = {10.1016/j.jgr.2023.07.005},
issn = {1226-8453},
year = {2023},
date = {2023-11-01},
urldate = {2025-05-02},
volume = {47},
number = {6},
pages = {694–705},
abstract = {Panax ginseng Meyer is a traditional Chinese medicine that is widely used as tonic in Asia. The main pharmacologically active components of ginseng are the dammarane-type ginsenosides, which have been shown to have anti-cancer, anti-inflammatory, immunoregulatory, neuroprotective, and metabolic regulatory activities. Moreover, some of ginsenosides (eg, Rh2 and Rg3) have been developed into nutraceuticals. However, the utilization of ginsenosides in clinic is restrictive due to poor permeability in cells and low bioavailability in human body. Obviously, the dammarane skeleton and glycosyls of ginsenosides are responsible for these limitations. Therefore, improving the oral bioavailability of ginsenosides has become a pressing issue. Here, based on the structures of ginsenosides, we summarized the understanding of the factors affecting the oral bioavailability of ginsenosides, introduced the methods to enhance the oral bioavailability and proposed the future perspectives on improving the oral bioavailability of ginsenosides.},
keywords = {ginsenoside, Gut microbiota, oral bioavailability, P-glycoprotein},
pubstate = {published},
tppubtype = {article}
}
Panax ginseng Meyer is a traditional Chinese medicine that is widely used as tonic in Asia. The main pharmacologically active components of ginseng are the dammarane-type ginsenosides, which have been shown to have anti-cancer, anti-inflammatory, immunoregulatory, neuroprotective, and metabolic regulatory activities. Moreover, some of ginsenosides (eg, Rh2 and Rg3) have been developed into nutraceuticals. However, the utilization of ginsenosides in clinic is restrictive due to poor permeability in cells and low bioavailability in human body. Obviously, the dammarane skeleton and glycosyls of ginsenosides are responsible for these limitations. Therefore, improving the oral bioavailability of ginsenosides has become a pressing issue. Here, based on the structures of ginsenosides, we summarized the understanding of the factors affecting the oral bioavailability of ginsenosides, introduced the methods to enhance the oral bioavailability and proposed the future perspectives on improving the oral bioavailability of ginsenosides.
Kus, Marta; Ibragimow, Izabela; Piotrowska-Kempisty, Hanna
Caco-2 Cell Line Standardization with Pharmaceutical Requirements and In Vitro Model Suitability for Permeability Assays Journal Article
In: vol. 15, no. 11, pp. 2523, 2023, ISSN: 1999-4923.
Abstract | Links | BibTeX | Tags:
@article{kus_caco-2_2023,
title = {Caco-2 Cell Line Standardization with Pharmaceutical Requirements and In Vitro Model Suitability for Permeability Assays},
author = {Marta Kus and Izabela Ibragimow and Hanna Piotrowska-Kempisty},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10674574/},
doi = {10.3390/pharmaceutics15112523},
issn = {1999-4923},
year = {2023},
date = {2023-10-24},
urldate = {2024-06-06},
volume = {15},
number = {11},
pages = {2523},
abstract = {The Caco-2 cell line derived from human colon carcinoma is commonly used to assess the permeability of compounds in in vitro conditions. Due to the significant increase in permeability studies using the Caco-2 cell line in recent years, the need to standardize this biological model seems necessary. The pharmaceutical requirements define only the acceptance criteria for the validation of the Caco-2 cell line and do not specify the protocol for its implementation. Therefore, the aim of this study is to review the conditions for permeability studies across the Caco-2 monolayer reported in the available literature concerning validation guidelines. We summarized the main aspects affecting the validation process of the Caco-2 cell line, including the culture conditions, cytotoxicity, cell differentiation process, and monolayer transport conditions, and the main conclusions may be useful in developing individual methods for preparing the cell line for validation purposes and further permeability research.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The Caco-2 cell line derived from human colon carcinoma is commonly used to assess the permeability of compounds in in vitro conditions. Due to the significant increase in permeability studies using the Caco-2 cell line in recent years, the need to standardize this biological model seems necessary. The pharmaceutical requirements define only the acceptance criteria for the validation of the Caco-2 cell line and do not specify the protocol for its implementation. Therefore, the aim of this study is to review the conditions for permeability studies across the Caco-2 monolayer reported in the available literature concerning validation guidelines. We summarized the main aspects affecting the validation process of the Caco-2 cell line, including the culture conditions, cytotoxicity, cell differentiation process, and monolayer transport conditions, and the main conclusions may be useful in developing individual methods for preparing the cell line for validation purposes and further permeability research.
Brügger, Michael David; Basler, Konrad
The diverse nature of intestinal fibroblasts in development, homeostasis, and disease Journal Article
In: vol. 33, no. 10, pp. 834–849, 2023, ISSN: 0962-8924, 1879-3088, (Publisher: Elsevier).
Links | BibTeX | Tags: colorectal cancer, inflammatory disease, intestinal development, intestinal fibroblasts, intestinal homeostasis, single-cell transcriptomic analysis
@article{brugger_diverse_2023,
title = {The diverse nature of intestinal fibroblasts in development, homeostasis, and disease},
author = {Michael David Brügger and Konrad Basler},
url = {https://www.cell.com/trends/cell-biology/abstract/S0962-8924(23)00048-X},
doi = {10.1016/j.tcb.2023.03.007},
issn = {0962-8924, 1879-3088},
year = {2023},
date = {2023-10-01},
urldate = {2024-07-02},
volume = {33},
number = {10},
pages = {834–849},
note = {Publisher: Elsevier},
keywords = {colorectal cancer, inflammatory disease, intestinal development, intestinal fibroblasts, intestinal homeostasis, single-cell transcriptomic analysis},
pubstate = {published},
tppubtype = {article}
}
Pike, Colleen M.; Zwarycz, Bailey; McQueen, Bryan E.; Castillo, Mariana; Barron, Catherine; Morowitz, Jeremy M.; Levi, James A.; Phadke, Dhiral; Balik-Meisner, Michele; Mav, Deepak; Shah, Ruchir; Glasspoole, Danielle L. Cunningham; Laetham, Ron; Thelin, William; Bunger, Maureen K.; Boazak, Elizabeth M.
Characterization and optimization of variability in a human colonic epithelium culture model Miscellaneous
2023, (Pages: 2023.09.22.559007 Section: New Results).
Abstract | Links | BibTeX | Tags:
@misc{pike_characterization_2023,
title = {Characterization and optimization of variability in a human colonic epithelium culture model},
author = {Colleen M. Pike and Bailey Zwarycz and Bryan E. McQueen and Mariana Castillo and Catherine Barron and Jeremy M. Morowitz and James A. Levi and Dhiral Phadke and Michele Balik-Meisner and Deepak Mav and Ruchir Shah and Danielle L. Cunningham Glasspoole and Ron Laetham and William Thelin and Maureen K. Bunger and Elizabeth M. Boazak},
url = {https://www.biorxiv.org/content/10.1101/2023.09.22.559007v1},
doi = {10.1101/2023.09.22.559007},
year = {2023},
date = {2023-09-22},
urldate = {2023-09-27},
publisher = {bioRxiv},
abstract = {Animal models have historically been poor preclinical predictors of gastrointestinal (GI) directed therapeutic efficacy and drug-induced GI toxicity. Human stem and primary cell-derived culture systems are a major focus of efforts to create biologically relevant models that enhance preclinical predictive value of intestinal efficacy and toxicity. The inherent variability in stem-cell-based complex cultures makes development of useful models a challenge; the stochastic nature of stem-cell differentiation interferes with the ability to build and validate robust, reproducible assays that query drug responses and pharmacokinetics. In this study, we aimed to characterize and reduce potential sources of variability in a complex stem cell-derived intestinal epithelium model, termed RepliGut® Planar, across cells from multiple human donors, cell lots, and passage numbers. Assessment criteria included barrier formation and integrity, gene expression, and cytokine responses. Gene expression and culture metric analyses revealed that controlling for stem/progenitor-cell passage number reduces variability and maximizes physiological relevance of the model. After optimizing passage number, donor-specific differences in cytokine responses were observed in a case study, suggesting biologic variability is observable in cell cultures derived from multiple human sources. Our findings highlight key considerations for designing assays that can be applied to additional primary-cell derived systems, as well as establish utility of the RepliGut® Planar platform for robust development of human-predictive drug-response assays.},
note = {Pages: 2023.09.22.559007
Section: New Results},
keywords = {},
pubstate = {published},
tppubtype = {misc}
}
Animal models have historically been poor preclinical predictors of gastrointestinal (GI) directed therapeutic efficacy and drug-induced GI toxicity. Human stem and primary cell-derived culture systems are a major focus of efforts to create biologically relevant models that enhance preclinical predictive value of intestinal efficacy and toxicity. The inherent variability in stem-cell-based complex cultures makes development of useful models a challenge; the stochastic nature of stem-cell differentiation interferes with the ability to build and validate robust, reproducible assays that query drug responses and pharmacokinetics. In this study, we aimed to characterize and reduce potential sources of variability in a complex stem cell-derived intestinal epithelium model, termed RepliGut® Planar, across cells from multiple human donors, cell lots, and passage numbers. Assessment criteria included barrier formation and integrity, gene expression, and cytokine responses. Gene expression and culture metric analyses revealed that controlling for stem/progenitor-cell passage number reduces variability and maximizes physiological relevance of the model. After optimizing passage number, donor-specific differences in cytokine responses were observed in a case study, suggesting biologic variability is observable in cell cultures derived from multiple human sources. Our findings highlight key considerations for designing assays that can be applied to additional primary-cell derived systems, as well as establish utility of the RepliGut® Planar platform for robust development of human-predictive drug-response assays.
Mehta, M.; Polli, J. E.; Seo, P.; Bhoopathy, S.; Berginc, K.; Kristan, K.; Cook, J.; Dressman, J. B.; Mandula, H.; Munshi, U.; Shanker, R.; Volpe, D. A.; Gordon, J.; Veerasingham, S.; Welink, J.; Almeida, S.; Gonzalez, P.; Painter, D.; Tsang, Y. C.; Vaidyanathan, J.; Velagapudi, R.
Drug Permeability - Best Practices for Biopharmaceutics Classification System (BCS)-Based Biowaivers: A workshop Summary Report Journal Article
In: vol. 112, no. 7, pp. 1749–1762, 2023, ISSN: 0022-3549.
Abstract | Links | BibTeX | Tags: {BCS}, {ICH} M9, biopharmaceutics classification system, Biowaiver, Caco2 cell line, permeability
@article{mehta_drug_2023,
title = {Drug Permeability - Best Practices for Biopharmaceutics Classification System (BCS)-Based Biowaivers: A workshop Summary Report},
author = {M. Mehta and J. E. Polli and P. Seo and S. Bhoopathy and K. Berginc and K. Kristan and J. Cook and J. B. Dressman and H. Mandula and U. Munshi and R. Shanker and D. A. Volpe and J. Gordon and S. Veerasingham and J. Welink and S. Almeida and P. Gonzalez and D. Painter and Y. C. Tsang and J. Vaidyanathan and R. Velagapudi},
url = {https://www.sciencedirect.com/science/article/pii/S0022354923001818},
doi = {10.1016/j.xphs.2023.04.016},
issn = {0022-3549},
year = {2023},
date = {2023-07-01},
urldate = {2024-06-26},
volume = {112},
number = {7},
pages = {1749–1762},
abstract = {The workshop “Drug Permeability - Best Practices for Biopharmaceutics Classification System (BCS) Based Biowaivers” was held virtually on December 6, 2021, organized by the University of Maryland Center of Excellence in Regulatory Science and Innovation (M-CERSI), and the Food and Drug Administration (FDA). The workshop focused on the industrial, academic, and regulatory experiences in generating and evaluating permeability data, with the aim to further facilitate implementation of the BCS and efficient development of high-quality drug products globally. As the first international permeability workshop since the BCS based biowaivers was finalized as the ICH M9 guideline, the workshop included lectures, panel discussions, and breakout sessions. Lecture and panel discussion topics covered case studies at IND, NDA, and ANDA stages, typical deficiencies relating to permeability assessment supporting BCS biowaiver, types of evidence that are available to demonstrate high permeability, method suitability of a permeability assay, impact of excipients, importance of global acceptance of permeability methods, opportunities to expand the use of biowaivers (e.g. non-Caco-2 cell lines, totality-of-evidence approach to demonstrate high permeability) and future of permeability testing. Breakout sessions focused on 1) in vitro and in silico intestinal permeability methods; 2) potential excipient effects on permeability and; 3) use of label and literature data to designate permeability class.},
keywords = {{BCS}, {ICH} M9, biopharmaceutics classification system, Biowaiver, Caco2 cell line, permeability},
pubstate = {published},
tppubtype = {article}
}
The workshop “Drug Permeability - Best Practices for Biopharmaceutics Classification System (BCS) Based Biowaivers” was held virtually on December 6, 2021, organized by the University of Maryland Center of Excellence in Regulatory Science and Innovation (M-CERSI), and the Food and Drug Administration (FDA). The workshop focused on the industrial, academic, and regulatory experiences in generating and evaluating permeability data, with the aim to further facilitate implementation of the BCS and efficient development of high-quality drug products globally. As the first international permeability workshop since the BCS based biowaivers was finalized as the ICH M9 guideline, the workshop included lectures, panel discussions, and breakout sessions. Lecture and panel discussion topics covered case studies at IND, NDA, and ANDA stages, typical deficiencies relating to permeability assessment supporting BCS biowaiver, types of evidence that are available to demonstrate high permeability, method suitability of a permeability assay, impact of excipients, importance of global acceptance of permeability methods, opportunities to expand the use of biowaivers (e.g. non-Caco-2 cell lines, totality-of-evidence approach to demonstrate high permeability) and future of permeability testing. Breakout sessions focused on 1) in vitro and in silico intestinal permeability methods; 2) potential excipient effects on permeability and; 3) use of label and literature data to designate permeability class.
Schiffman, Susan S.; Scholl, Elizabeth H.; Furey, Terrence S.; Nagle, H. Troy
Toxicological and pharmacokinetic properties of sucralose-6-acetate and its parent sucralose: in vitro screening assays Journal Article
In: vol. 0, no. 0, pp. 1–35, 2023, ISSN: 1093-7404, (Publisher: Taylor & Francis _eprint: https://doi.org/10.1080/10937404.2023.2213903).
Abstract | Links | BibTeX | Tags: Gene Expression, genotoxicity, intestinal barrier, Sucralose, sucralose-6-acetate
@article{schiffman_toxicological_2023,
title = {Toxicological and pharmacokinetic properties of sucralose-6-acetate and its parent sucralose: in vitro screening assays},
author = {Susan S. Schiffman and Elizabeth H. Scholl and Terrence S. Furey and H. Troy Nagle},
url = {https://doi.org/10.1080/10937404.2023.2213903},
doi = {10.1080/10937404.2023.2213903},
issn = {1093-7404},
year = {2023},
date = {2023-05-29},
urldate = {2023-06-02},
volume = {0},
number = {0},
pages = {1–35},
abstract = {The purpose of this study was to determine the toxicological and pharmacokinetic properties of sucralose-6-acetate, a structural analog of the artificial sweetener sucralose. Sucralose-6-acetate is an intermediate and impurity in the manufacture of sucralose, and recent commercial sucralose samples were found to contain up to 0.67% sucralose-6-acetate. Studies in a rodent model found that sucralose-6-acetate is also present in fecal samples with levels up to 10% relative to sucralose which suggest that sucralose is also acetylated in the intestines. A MultiFlow® assay, a high-throughput genotoxicity screening tool, and a micronucleus (MN) test that detects cytogenetic damage both indicated that sucralose-6-acetate is genotoxic. The mechanism of action was classified as clastogenic (produces DNA strand breaks) using the MultiFlow® assay. The amount of sucralose-6-acetate in a single daily sucralose-sweetened drink might far exceed the threshold of toxicological concern for genotoxicity (TTCgenotox) of 0.15 µg/person/day. The RepliGut® System was employed to expose human intestinal epithelium to sucralose-6-acetate and sucralose, and an RNA-seq analysis was performed to determine gene expression induced by these exposures. Sucralose-6-acetate significantly increased the expression of genes associated with inflammation, oxidative stress, and cancer with greatest expression for the metallothionein 1 G gene (MT1G). Measurements of transepithelial electrical resistance (TEER) and permeability in human transverse colon epithelium indicated that sucralose-6-acetate and sucralose both impaired intestinal barrier integrity. Sucralose-6-acetate also inhibited two members of the cytochrome P450 family (CYP1A2 and CYP2C19). Overall, the toxicological and pharmacokinetic findings for sucralose-6-acetate raise significant health concerns regarding the safety and regulatory status of sucralose itself.},
note = {Publisher: Taylor & Francis
_eprint: https://doi.org/10.1080/10937404.2023.2213903},
keywords = {Gene Expression, genotoxicity, intestinal barrier, Sucralose, sucralose-6-acetate},
pubstate = {published},
tppubtype = {article}
}
The purpose of this study was to determine the toxicological and pharmacokinetic properties of sucralose-6-acetate, a structural analog of the artificial sweetener sucralose. Sucralose-6-acetate is an intermediate and impurity in the manufacture of sucralose, and recent commercial sucralose samples were found to contain up to 0.67% sucralose-6-acetate. Studies in a rodent model found that sucralose-6-acetate is also present in fecal samples with levels up to 10% relative to sucralose which suggest that sucralose is also acetylated in the intestines. A MultiFlow® assay, a high-throughput genotoxicity screening tool, and a micronucleus (MN) test that detects cytogenetic damage both indicated that sucralose-6-acetate is genotoxic. The mechanism of action was classified as clastogenic (produces DNA strand breaks) using the MultiFlow® assay. The amount of sucralose-6-acetate in a single daily sucralose-sweetened drink might far exceed the threshold of toxicological concern for genotoxicity (TTCgenotox) of 0.15 µg/person/day. The RepliGut® System was employed to expose human intestinal epithelium to sucralose-6-acetate and sucralose, and an RNA-seq analysis was performed to determine gene expression induced by these exposures. Sucralose-6-acetate significantly increased the expression of genes associated with inflammation, oxidative stress, and cancer with greatest expression for the metallothionein 1 G gene (MT1G). Measurements of transepithelial electrical resistance (TEER) and permeability in human transverse colon epithelium indicated that sucralose-6-acetate and sucralose both impaired intestinal barrier integrity. Sucralose-6-acetate also inhibited two members of the cytochrome P450 family (CYP1A2 and CYP2C19). Overall, the toxicological and pharmacokinetic findings for sucralose-6-acetate raise significant health concerns regarding the safety and regulatory status of sucralose itself.
Zuo, Li; Kuo, Wei-Ting; Cao, Feng; Chanez-Paredes, Sandra D.; Zeve, Daniel; Mannam, Prabhath; Jean-François, Léa; Day, Anne; Graham, W. Vallen; Sweat, Yan Y.; Shashikanth, Nitesh; Breault, David T.; Turner, Jerrold R.
In: vol. 72, no. 5, pp. 870–881, 2023, ISSN: 0017-5749, 1468-3288, (Publisher: BMJ Publishing Group Section: Inflammatory bowel disease).
Abstract | Links | BibTeX | Tags: {IBD} basic research, {TNF}, actin cytoskeleton, intestinal barrier function, intestinal stem cell
@article{zuo_tacrolimus-binding_2023,
title = {Tacrolimus-binding protein FKBP8 directs myosin light chain kinase-dependent barrier regulation and is a potential therapeutic target in Crohn’s disease},
author = {Li Zuo and Wei-Ting Kuo and Feng Cao and Sandra D. Chanez-Paredes and Daniel Zeve and Prabhath Mannam and Léa Jean-François and Anne Day and W. Vallen Graham and Yan Y. Sweat and Nitesh Shashikanth and David T. Breault and Jerrold R. Turner},
url = {https://gut.bmj.com/content/72/5/870},
doi = {10.1136/gutjnl-2021-326534},
issn = {0017-5749, 1468-3288},
year = {2023},
date = {2023-05-01},
urldate = {2023-04-27},
volume = {72},
number = {5},
pages = {870–881},
abstract = {Objective Intestinal barrier loss is a Crohn’s disease (CD) risk factor. This may be related to increased expression and enzymatic activation of myosin light chain kinase 1 (MLCK1), which increases intestinal paracellular permeability and correlates with CD severity. Moreover, preclinical studies have shown that MLCK1 recruitment to cell junctions is required for tumour necrosis factor (TNF)-induced barrier loss as well as experimental inflammatory bowel disease progression. We sought to define mechanisms of MLCK1 recruitment and to target this process pharmacologically.
Design Protein interactions between FK506 binding protein 8 (FKBP8) and MLCK1 were assessed in vitro. Transgenic and knockout intestinal epithelial cell lines, human intestinal organoids, and mice were used as preclinical models. Discoveries were validated in biopsies from patients with CD and control subjects.
Results MLCK1 interacted specifically with the tacrolimus-binding FKBP8 PPI domain. Knockout or dominant negative FKBP8 expression prevented TNF-induced MLCK1 recruitment and barrier loss in vitro. MLCK1-FKBP8 binding was blocked by tacrolimus, which reversed TNF-induced MLCK1-FKBP8 interactions, MLCK1 recruitment and barrier loss in vitro and in vivo. Biopsies of patient with CD demonstrated increased numbers of MLCK1-FKBP8 interactions at intercellular junctions relative to control subjects.
Conclusion Binding to FKBP8, which can be blocked by tacrolimus, is required for MLCK1 recruitment to intercellular junctions and downstream events leading to immune-mediated barrier loss. The observed increases in MLCK1 activity, MLCK1 localisation at cell junctions and perijunctional MLCK1-FKBP8 interactions in CD suggest that targeting this process may be therapeutic in human disease. These new insights into mechanisms of disease-associated barrier loss provide a critical foundation for therapeutic exploitation of FKBP8-MLCK1 interactions.},
note = {Publisher: BMJ Publishing Group
Section: Inflammatory bowel disease},
keywords = {{IBD} basic research, {TNF}, actin cytoskeleton, intestinal barrier function, intestinal stem cell},
pubstate = {published},
tppubtype = {article}
}
Objective Intestinal barrier loss is a Crohn’s disease (CD) risk factor. This may be related to increased expression and enzymatic activation of myosin light chain kinase 1 (MLCK1), which increases intestinal paracellular permeability and correlates with CD severity. Moreover, preclinical studies have shown that MLCK1 recruitment to cell junctions is required for tumour necrosis factor (TNF)-induced barrier loss as well as experimental inflammatory bowel disease progression. We sought to define mechanisms of MLCK1 recruitment and to target this process pharmacologically.
Design Protein interactions between FK506 binding protein 8 (FKBP8) and MLCK1 were assessed in vitro. Transgenic and knockout intestinal epithelial cell lines, human intestinal organoids, and mice were used as preclinical models. Discoveries were validated in biopsies from patients with CD and control subjects.
Results MLCK1 interacted specifically with the tacrolimus-binding FKBP8 PPI domain. Knockout or dominant negative FKBP8 expression prevented TNF-induced MLCK1 recruitment and barrier loss in vitro. MLCK1-FKBP8 binding was blocked by tacrolimus, which reversed TNF-induced MLCK1-FKBP8 interactions, MLCK1 recruitment and barrier loss in vitro and in vivo. Biopsies of patient with CD demonstrated increased numbers of MLCK1-FKBP8 interactions at intercellular junctions relative to control subjects.
Conclusion Binding to FKBP8, which can be blocked by tacrolimus, is required for MLCK1 recruitment to intercellular junctions and downstream events leading to immune-mediated barrier loss. The observed increases in MLCK1 activity, MLCK1 localisation at cell junctions and perijunctional MLCK1-FKBP8 interactions in CD suggest that targeting this process may be therapeutic in human disease. These new insights into mechanisms of disease-associated barrier loss provide a critical foundation for therapeutic exploitation of FKBP8-MLCK1 interactions.
Design Protein interactions between FK506 binding protein 8 (FKBP8) and MLCK1 were assessed in vitro. Transgenic and knockout intestinal epithelial cell lines, human intestinal organoids, and mice were used as preclinical models. Discoveries were validated in biopsies from patients with CD and control subjects.
Results MLCK1 interacted specifically with the tacrolimus-binding FKBP8 PPI domain. Knockout or dominant negative FKBP8 expression prevented TNF-induced MLCK1 recruitment and barrier loss in vitro. MLCK1-FKBP8 binding was blocked by tacrolimus, which reversed TNF-induced MLCK1-FKBP8 interactions, MLCK1 recruitment and barrier loss in vitro and in vivo. Biopsies of patient with CD demonstrated increased numbers of MLCK1-FKBP8 interactions at intercellular junctions relative to control subjects.
Conclusion Binding to FKBP8, which can be blocked by tacrolimus, is required for MLCK1 recruitment to intercellular junctions and downstream events leading to immune-mediated barrier loss. The observed increases in MLCK1 activity, MLCK1 localisation at cell junctions and perijunctional MLCK1-FKBP8 interactions in CD suggest that targeting this process may be therapeutic in human disease. These new insights into mechanisms of disease-associated barrier loss provide a critical foundation for therapeutic exploitation of FKBP8-MLCK1 interactions.
Horowitz, Arie; Chanez-Paredes, Sandra D.; Haest, Xenia; Turner, Jerrold R.
Paracellular permeability and tight junction regulation in gut health and disease Journal Article
In: pp. 1–16, 2023, ISSN: 1759-5053, (Publisher: Nature Publishing Group).
Abstract | Links | BibTeX | Tags: Gastroenterology, Gastrointestinal system, inflammatory bowel disease
@article{horowitz_paracellular_2023,
title = {Paracellular permeability and tight junction regulation in gut health and disease},
author = {Arie Horowitz and Sandra D. Chanez-Paredes and Xenia Haest and Jerrold R. Turner},
url = {https://www.nature.com/articles/s41575-023-00766-3},
doi = {10.1038/s41575-023-00766-3},
issn = {1759-5053},
year = {2023},
date = {2023-04-25},
urldate = {2023-04-27},
pages = {1–16},
abstract = {Epithelial tight junctions define the paracellular permeability of the intestinal barrier. Molecules can cross the tight junctions via two distinct size-selective and charge-selective paracellular pathways: the pore pathway and the leak pathway. These can be distinguished by their selectivities and differential regulation by immune cells. However, permeability increases measured in most studies are secondary to epithelial damage, which allows non-selective flux via the unrestricted pathway. Restoration of increased unrestricted pathway permeability requires mucosal healing. By contrast, tight junction barrier loss can be reversed by targeted interventions. Specific approaches are needed to restore pore pathway or leak pathway permeability increases. Recent studies have used preclinical disease models to demonstrate the potential of pore pathway or leak pathway barrier restoration in disease. In this Review, we focus on the two paracellular flux pathways that are dependent on the tight junction. We discuss the latest evidence that highlights tight junction components, structures and regulatory mechanisms, their impact on gut health and disease, and opportunities for therapeutic intervention.},
note = {Publisher: Nature Publishing Group},
keywords = {Gastroenterology, Gastrointestinal system, inflammatory bowel disease},
pubstate = {published},
tppubtype = {article}
}
Epithelial tight junctions define the paracellular permeability of the intestinal barrier. Molecules can cross the tight junctions via two distinct size-selective and charge-selective paracellular pathways: the pore pathway and the leak pathway. These can be distinguished by their selectivities and differential regulation by immune cells. However, permeability increases measured in most studies are secondary to epithelial damage, which allows non-selective flux via the unrestricted pathway. Restoration of increased unrestricted pathway permeability requires mucosal healing. By contrast, tight junction barrier loss can be reversed by targeted interventions. Specific approaches are needed to restore pore pathway or leak pathway permeability increases. Recent studies have used preclinical disease models to demonstrate the potential of pore pathway or leak pathway barrier restoration in disease. In this Review, we focus on the two paracellular flux pathways that are dependent on the tight junction. We discuss the latest evidence that highlights tight junction components, structures and regulatory mechanisms, their impact on gut health and disease, and opportunities for therapeutic intervention.
Sarma, Sudeep; Catella, Carly M.; Pedro, Ellyce T. San; Xiao, Xingqing; Durmusoglu, Deniz; Menegatti, Stefano; Crook, Nathan; Magness, Scott T.; Hall, Carol K.
Design of 8-mer Peptides that Block Clostridioides difficile Toxin A in Intestinal Cells Journal Article
In: pp. 2023.01.10.523493, 2023.
Abstract | Links | BibTeX | Tags:
@article{sarma_design_2023,
title = {Design of 8-mer Peptides that Block Clostridioides difficile Toxin A in Intestinal Cells},
author = {Sudeep Sarma and Carly M. Catella and Ellyce T. San Pedro and Xingqing Xiao and Deniz Durmusoglu and Stefano Menegatti and Nathan Crook and Scott T. Magness and Carol K. Hall},
doi = {10.1101/2023.01.10.523493},
year = {2023},
date = {2023-01-12},
pages = {2023.01.10.523493},
abstract = {Clostridioides difficile ( C. diff .) is a bacterium that causes severe diarrhea and inflammation of the colon. The pathogenicity of C. diff . infection is derived from two major toxins, toxins A (TcdA) and B (TcdB). Peptide inhibitors that can be delivered to the gut to inactivate these toxins are an attractive therapeutic strategy. In this work, we present a new approach that combines a pep tide b inding d esign algorithm (PepBD), molecular-level simulations, rapid screening of candidate peptides for toxin binding, a primary human cell-based assay, and surface plasmon resonance (SPR) measurements to develop peptide inhibitors that block the glucosyltransferase activity of TcdA by targeting its glucosyltransferase domain (GTD). Using PepBD and explicit-solvent molecular dynamics simulations, we identified seven candidate peptides, SA1-SA7. These peptides were selected for specific TcdA GTD binding through a custom solid-phase peptide screening system, which eliminated the weaker inhibitors SA5-SA7. The efficacies of SA1-SA4 were then tested using a trans-epithelial electrical resistance (TEER) assay on monolayers of the human gut epithelial culture model. One peptide, SA1, was found to block TcdA toxicity in primary-derived human jejunum (small intestinal) and colon (large intestinal) epithelial cells. SA1 bound TcdA with a K D of 56.1 ± 29.8 nM as measured by surface plasmon resonance (SPR).
SIGNIFICANCE STATEMENT: Infections by Clostridioides difficile , a bacterium that targets the large intestine (colon), impact a significant number of people worldwide. Bacterial colonization is mediated by two exotoxins: toxins A and B. Short peptides that can inhibit the biocatalytic activity of these toxins represent a promising strategy to prevent and treat C. diff . infection. We describe an approach that combines a Peptide B inding D esign (PepBD) algorithm, molecular-level simulations, a rapid screening assay to evaluate peptide:toxin binding, a primary human cell-based assay, and surface plasmon resonance (SPR) measurements to develop peptide inhibitors that block Toxin A in small intestinal and colon epithelial cells. Importantly, our designed peptide, SA1, bound toxin A with nanomolar affinity and blocked toxicity in colon cells.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Clostridioides difficile ( C. diff .) is a bacterium that causes severe diarrhea and inflammation of the colon. The pathogenicity of C. diff . infection is derived from two major toxins, toxins A (TcdA) and B (TcdB). Peptide inhibitors that can be delivered to the gut to inactivate these toxins are an attractive therapeutic strategy. In this work, we present a new approach that combines a pep tide b inding d esign algorithm (PepBD), molecular-level simulations, rapid screening of candidate peptides for toxin binding, a primary human cell-based assay, and surface plasmon resonance (SPR) measurements to develop peptide inhibitors that block the glucosyltransferase activity of TcdA by targeting its glucosyltransferase domain (GTD). Using PepBD and explicit-solvent molecular dynamics simulations, we identified seven candidate peptides, SA1-SA7. These peptides were selected for specific TcdA GTD binding through a custom solid-phase peptide screening system, which eliminated the weaker inhibitors SA5-SA7. The efficacies of SA1-SA4 were then tested using a trans-epithelial electrical resistance (TEER) assay on monolayers of the human gut epithelial culture model. One peptide, SA1, was found to block TcdA toxicity in primary-derived human jejunum (small intestinal) and colon (large intestinal) epithelial cells. SA1 bound TcdA with a K D of 56.1 ± 29.8 nM as measured by surface plasmon resonance (SPR).
SIGNIFICANCE STATEMENT: Infections by Clostridioides difficile , a bacterium that targets the large intestine (colon), impact a significant number of people worldwide. Bacterial colonization is mediated by two exotoxins: toxins A and B. Short peptides that can inhibit the biocatalytic activity of these toxins represent a promising strategy to prevent and treat C. diff . infection. We describe an approach that combines a Peptide B inding D esign (PepBD) algorithm, molecular-level simulations, a rapid screening assay to evaluate peptide:toxin binding, a primary human cell-based assay, and surface plasmon resonance (SPR) measurements to develop peptide inhibitors that block Toxin A in small intestinal and colon epithelial cells. Importantly, our designed peptide, SA1, bound toxin A with nanomolar affinity and blocked toxicity in colon cells.
SIGNIFICANCE STATEMENT: Infections by Clostridioides difficile , a bacterium that targets the large intestine (colon), impact a significant number of people worldwide. Bacterial colonization is mediated by two exotoxins: toxins A and B. Short peptides that can inhibit the biocatalytic activity of these toxins represent a promising strategy to prevent and treat C. diff . infection. We describe an approach that combines a Peptide B inding D esign (PepBD) algorithm, molecular-level simulations, a rapid screening assay to evaluate peptide:toxin binding, a primary human cell-based assay, and surface plasmon resonance (SPR) measurements to develop peptide inhibitors that block Toxin A in small intestinal and colon epithelial cells. Importantly, our designed peptide, SA1, bound toxin A with nanomolar affinity and blocked toxicity in colon cells.
McCoubrey, Laura E.; Favaron, Alessia; Awad, Atheer; Orlu, Mine; Gaisford, Simon; Basit, Abdul W.
Colonic drug delivery: Formulating the next generation of colon-targeted therapeutics Journal Article
In: vol. 353, pp. 1107–1126, 2023, ISSN: 0168-3659.
Abstract | Links | BibTeX | Tags: Artificial intelligence and machine learning, Drug delivery systems and the large intestine, Gastrointestinal microbiota and microbiome, Inflammatory bowel disease and drug products, Oral biologics and biopharmaceuticals, Targeting the colon
@article{mccoubrey_colonic_2023,
title = {Colonic drug delivery: Formulating the next generation of colon-targeted therapeutics},
author = {Laura E. McCoubrey and Alessia Favaron and Atheer Awad and Mine Orlu and Simon Gaisford and Abdul W. Basit},
url = {https://www.sciencedirect.com/science/article/pii/S0168365922008434},
doi = {10.1016/j.jconrel.2022.12.029},
issn = {0168-3659},
year = {2023},
date = {2023-01-01},
urldate = {2024-04-02},
volume = {353},
pages = {1107–1126},
abstract = {Colonic drug delivery can facilitate access to unique therapeutic targets and has the potential to enhance drug bioavailability whilst reducing off-target effects. Delivering drugs to the colon requires considered formulation development, as both oral and rectal dosage forms can encounter challenges if the colon’s distinct physiological environment is not appreciated. As the therapeutic opportunities surrounding colonic drug delivery multiply, the success of novel pharmaceuticals lies in their design. This review provides a modern insight into the key parameters determining the effective design and development of colon-targeted medicines. Influential physiological features governing the release, dissolution, stability, and absorption of drugs in the colon are first discussed, followed by an overview of the most reliable colon-targeted formulation strategies. Finally, the most appropriate in vitro, in vivo, and in silico preclinical investigations are presented, with the goal of inspiring strategic development of new colon-targeted therapeutics.},
keywords = {Artificial intelligence and machine learning, Drug delivery systems and the large intestine, Gastrointestinal microbiota and microbiome, Inflammatory bowel disease and drug products, Oral biologics and biopharmaceuticals, Targeting the colon},
pubstate = {published},
tppubtype = {article}
}
Colonic drug delivery can facilitate access to unique therapeutic targets and has the potential to enhance drug bioavailability whilst reducing off-target effects. Delivering drugs to the colon requires considered formulation development, as both oral and rectal dosage forms can encounter challenges if the colon’s distinct physiological environment is not appreciated. As the therapeutic opportunities surrounding colonic drug delivery multiply, the success of novel pharmaceuticals lies in their design. This review provides a modern insight into the key parameters determining the effective design and development of colon-targeted medicines. Influential physiological features governing the release, dissolution, stability, and absorption of drugs in the colon are first discussed, followed by an overview of the most reliable colon-targeted formulation strategies. Finally, the most appropriate in vitro, in vivo, and in silico preclinical investigations are presented, with the goal of inspiring strategic development of new colon-targeted therapeutics.
2022
Bolster, Doug; Chae, Lee; van Klinken, Jan-Willem; Kalgaonkar, Swati
Impact of selected novel plant bioactives on improvement of impaired gut barrier function using human primary cell intestinal epithelium: Journal Article
In: vol. 20, 2022, ISSN: 2637-8779.
Abstract | Links | BibTeX | Tags: Bioactives, Gut barrier function, Gut permeability, Hepatic nuclear factor 4α, N-Trans-caffeoyltyramine, N-Trans-feruloyltyramine
@article{bolster_impact_2022,
title = {Impact of selected novel plant bioactives on improvement of impaired gut barrier function using human primary cell intestinal epithelium:},
author = {Doug Bolster and Lee Chae and Jan-Willem van Klinken and Swati Kalgaonkar},
url = {http://www.isnff-jfb.com/index.php/JFB/article/view/301},
doi = {10.31665/JFB.2022.18324},
issn = {2637-8779},
year = {2022},
date = {2022-12-30},
urldate = {2023-06-02},
volume = {20},
abstract = {Gut barrier function is compromised in the obese state. The N-trans caffeoyltyramine (NCT) and N-trans feruloyltyramine (NFT), two naturally occurring bioactive compounds in hemp hulls, identified using in silico approaches, have the potential to improve gut barrier function and their effects were studied here in vitro. Proliferative human transverse colon epithelial cells were plated and co-cultured with tumor necrosis factor (TNF) along with NCT, NFT or NCT/NFT (2.2 ratio) post-differentiation, over a 48-hour period to induce inflammation and to observe the effects of NCT and NFT. A decrease in transepithelial electrical resistance (TEER) and increase in the intestinal permeability were observed with increased addition of TNF. Co-administration of NCT and NFT demonstrated a dose-dependent and statistically significant reversal of impaired TEER and intestinal permeability. NCT and NFT demonstrated a physiologically relevant reversal of impaired gut barrier function in the setting of inflammation via significant improvement in TEER and percent permeability.},
keywords = {Bioactives, Gut barrier function, Gut permeability, Hepatic nuclear factor 4α, N-Trans-caffeoyltyramine, N-Trans-feruloyltyramine},
pubstate = {published},
tppubtype = {article}
}
Gut barrier function is compromised in the obese state. The N-trans caffeoyltyramine (NCT) and N-trans feruloyltyramine (NFT), two naturally occurring bioactive compounds in hemp hulls, identified using in silico approaches, have the potential to improve gut barrier function and their effects were studied here in vitro. Proliferative human transverse colon epithelial cells were plated and co-cultured with tumor necrosis factor (TNF) along with NCT, NFT or NCT/NFT (2.2 ratio) post-differentiation, over a 48-hour period to induce inflammation and to observe the effects of NCT and NFT. A decrease in transepithelial electrical resistance (TEER) and increase in the intestinal permeability were observed with increased addition of TNF. Co-administration of NCT and NFT demonstrated a dose-dependent and statistically significant reversal of impaired TEER and intestinal permeability. NCT and NFT demonstrated a physiologically relevant reversal of impaired gut barrier function in the setting of inflammation via significant improvement in TEER and percent permeability.
Barbieri, Maria Antonietta; Viola, Anna; Cicala, Giuseppe; Spina, Edoardo; Fries, Walter
Effectiveness and Safety Profiles of Biological Therapies in Inflammatory Bowel Disease: Real Life Data from an Active Pharmacovigilance Project Journal Article
In: vol. 10, no. 12, pp. 3280, 2022, ISSN: 2227-9059.
Abstract | Links | BibTeX | Tags:
@article{barbieri_effectiveness_2022,
title = {Effectiveness and Safety Profiles of Biological Therapies in Inflammatory Bowel Disease: Real Life Data from an Active Pharmacovigilance Project},
author = {Maria Antonietta Barbieri and Anna Viola and Giuseppe Cicala and Edoardo Spina and Walter Fries},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9775949/},
doi = {10.3390/biomedicines10123280},
issn = {2227-9059},
year = {2022},
date = {2022-12-18},
urldate = {2023-05-05},
volume = {10},
number = {12},
pages = {3280},
abstract = {Post-marketing surveillance is essential to evaluate the risk/benefit profile of drugs; however, pharmacovigilance studies comparing persistence and safety of biologic therapies in patients with inflammatory bowel disease (IBD) are scant. The aim of this study was to prospectively investigate persistence together with safety profiles of biologics in a cohort of patients diagnosed with Crohn’s Disease (CD) or ulcerative colitis (UC) followed by the IBD unit of Messina and treated with infliximab (IFX), adalimumab (ADA), golimumab (GOL), vedolizumab (VED), and ustekinumab (UST) from 2017 through 2021. Descriptive and treatment persistence analyses with predictors for discontinuation and occurrence of adverse drug reactions (ADRs) were performed. A total of 675 IBD patients were enrolled. A higher persistence rate was noted for UST and ADA in the first year (83.8% and 83.1%, respectively) and for IFX in the fifth year of treatment (58.1%). GOL, VED, and UST—all used as second/third-line therapies—seemed to have a higher risk of non-persistence than IFX (in order HR: 2.19; CI 95%: 1.33–3.61, 1.45; 1.04–2.04, 2.25; 1.25–4.07) as well as switchers and those who had at least one ADR (18.1; 13.22–24.68 and 1.55; 1.20–1.99, respectively). The reported ADRs, which were generally mild–moderate, were largely known. However, real-world data should be implemented to further study undetected safety concerns, including risk of malignancy.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Post-marketing surveillance is essential to evaluate the risk/benefit profile of drugs; however, pharmacovigilance studies comparing persistence and safety of biologic therapies in patients with inflammatory bowel disease (IBD) are scant. The aim of this study was to prospectively investigate persistence together with safety profiles of biologics in a cohort of patients diagnosed with Crohn’s Disease (CD) or ulcerative colitis (UC) followed by the IBD unit of Messina and treated with infliximab (IFX), adalimumab (ADA), golimumab (GOL), vedolizumab (VED), and ustekinumab (UST) from 2017 through 2021. Descriptive and treatment persistence analyses with predictors for discontinuation and occurrence of adverse drug reactions (ADRs) were performed. A total of 675 IBD patients were enrolled. A higher persistence rate was noted for UST and ADA in the first year (83.8% and 83.1%, respectively) and for IFX in the fifth year of treatment (58.1%). GOL, VED, and UST—all used as second/third-line therapies—seemed to have a higher risk of non-persistence than IFX (in order HR: 2.19; CI 95%: 1.33–3.61, 1.45; 1.04–2.04, 2.25; 1.25–4.07) as well as switchers and those who had at least one ADR (18.1; 13.22–24.68 and 1.55; 1.20–1.99, respectively). The reported ADRs, which were generally mild–moderate, were largely known. However, real-world data should be implemented to further study undetected safety concerns, including risk of malignancy.
Schnur, Sabrina; Wahl, Vanessa; Metz, Julia K.; Gillmann, Jessica; Hans, Fabian; Rotermund, Katharina; Zäh, Ralf-Kilian; Brück, Dietmar A.; Schneider, Marc; Hittinger, Marius
Inflammatory bowel disease addressed by Caco-2 and monocyte-derived macrophages: an opportunity for an in vitro drug screening assay Journal Article
In: vol. 1, no. 4, pp. 365–383, 2022, ISSN: 2731-3441.
Abstract | Links | BibTeX | Tags: {IVIVC}, Cell-based co-culture, Chronic inflammation of {GI}-tract, Drug testing, Efficacy outcome pathways
@article{schnur_inflammatory_2022,
title = {Inflammatory bowel disease addressed by Caco-2 and monocyte-derived macrophages: an opportunity for an in vitro drug screening assay},
author = {Sabrina Schnur and Vanessa Wahl and Julia K. Metz and Jessica Gillmann and Fabian Hans and Katharina Rotermund and Ralf-Kilian Zäh and Dietmar A. Brück and Marc Schneider and Marius Hittinger},
url = {https://doi.org/10.1007/s44164-022-00035-8},
doi = {10.1007/s44164-022-00035-8},
issn = {2731-3441},
year = {2022},
date = {2022-11-01},
urldate = {2023-05-05},
volume = {1},
number = {4},
pages = {365–383},
abstract = {Inflammatory bowel disease (IBD) is a widespread disease, affecting a growing demographic. The treatment of chronic inflammation located in the GI-tract is dependent on the severity; therefore, the IBD treatment pyramid is commonly applied. Animal experimentation plays a key role for novel IBD drug development; nevertheless, it is ethically questionable and limited in its throughput. Reliable and valid in vitro assays offer the opportunity to overcome these limitations.},
keywords = {{IVIVC}, Cell-based co-culture, Chronic inflammation of {GI}-tract, Drug testing, Efficacy outcome pathways},
pubstate = {published},
tppubtype = {article}
}
Inflammatory bowel disease (IBD) is a widespread disease, affecting a growing demographic. The treatment of chronic inflammation located in the GI-tract is dependent on the severity; therefore, the IBD treatment pyramid is commonly applied. Animal experimentation plays a key role for novel IBD drug development; nevertheless, it is ethically questionable and limited in its throughput. Reliable and valid in vitro assays offer the opportunity to overcome these limitations.
da Silva Santos, Rafael Rocha; Ramos, Matheus Corrêa; Ferreira, Juliana Veloso; Gonçalves, José Eduardo; César, Isabela Costa
In: vol. 296, pp. 115480, 2022, ISSN: 0378-8741.
Abstract | Links | BibTeX | Tags: Caco-2 cell, Equilibrium solubility, G. Forst, Kava, Kavain, permeability
@article{santos_biopharmaceutical_2022-1,
title = {Biopharmaceutical evaluation of kavain in textitPiper methysticum G. Forst dried extract: Equilibrium solubility and intestinal permeability in Caco-2 cell model},
author = {Rafael Rocha da Silva Santos and Matheus Corrêa Ramos and Juliana Veloso Ferreira and José Eduardo Gonçalves and Isabela Costa César},
url = {https://www.sciencedirect.com/science/article/pii/S0378874122005190},
doi = {10.1016/j.jep.2022.115480},
issn = {0378-8741},
year = {2022},
date = {2022-10-05},
urldate = {2025-05-02},
volume = {296},
pages = {115480},
abstract = {Ethnopharmacological relevance
Piper methysticum G. Forst, popularly known as kava, is a traditional medicinal plant native from South Pacific islands and widely used to treat anxiety, depression and stress. The psychoactive properties are related to the kavalactones, mainly kavain.
Aim of the study
To evaluate the biopharmaceutical properties of synthetic kavain and when present in kava dried extracts by means of equilibrium solubility and intestinal permeability studies in the Caco-2 cell model.
Materials and methods
The equilibrium solubility of kavain was performed using a shake flask incubator at 37 °C in different media at physiological pH range (1.2–6.8). The intestinal permeability of kavain evaluated in Caco-2 cells in the presence and absence of the P-glycoprotein inhibitor verapamil. Kavain concentrations were determined by reversed phase high performance liquid chromatography (HPLC).
Results
HPLC methods were developed and fully validated for kavain quantitation. Kavain demonstrated low solubility and the pH of the aqueous media did not affect its solubility. Kavain was found to be highly permeable and efflux of kavain mediated by P-glycoprotein was not significant during intestinal permeation.
Conclusion
The results of biopharmaceutical studies provided useful information for predicting availability of kavain from the gastrointestinal tract and this compound was ranked as BCS Class II, exhibiting dissolution rate-limited absorption.},
keywords = {Caco-2 cell, Equilibrium solubility, G. Forst, Kava, Kavain, permeability},
pubstate = {published},
tppubtype = {article}
}
Ethnopharmacological relevance
Piper methysticum G. Forst, popularly known as kava, is a traditional medicinal plant native from South Pacific islands and widely used to treat anxiety, depression and stress. The psychoactive properties are related to the kavalactones, mainly kavain.
Aim of the study
To evaluate the biopharmaceutical properties of synthetic kavain and when present in kava dried extracts by means of equilibrium solubility and intestinal permeability studies in the Caco-2 cell model.
Materials and methods
The equilibrium solubility of kavain was performed using a shake flask incubator at 37 °C in different media at physiological pH range (1.2–6.8). The intestinal permeability of kavain evaluated in Caco-2 cells in the presence and absence of the P-glycoprotein inhibitor verapamil. Kavain concentrations were determined by reversed phase high performance liquid chromatography (HPLC).
Results
HPLC methods were developed and fully validated for kavain quantitation. Kavain demonstrated low solubility and the pH of the aqueous media did not affect its solubility. Kavain was found to be highly permeable and efflux of kavain mediated by P-glycoprotein was not significant during intestinal permeation.
Conclusion
The results of biopharmaceutical studies provided useful information for predicting availability of kavain from the gastrointestinal tract and this compound was ranked as BCS Class II, exhibiting dissolution rate-limited absorption.
Piper methysticum G. Forst, popularly known as kava, is a traditional medicinal plant native from South Pacific islands and widely used to treat anxiety, depression and stress. The psychoactive properties are related to the kavalactones, mainly kavain.
Aim of the study
To evaluate the biopharmaceutical properties of synthetic kavain and when present in kava dried extracts by means of equilibrium solubility and intestinal permeability studies in the Caco-2 cell model.
Materials and methods
The equilibrium solubility of kavain was performed using a shake flask incubator at 37 °C in different media at physiological pH range (1.2–6.8). The intestinal permeability of kavain evaluated in Caco-2 cells in the presence and absence of the P-glycoprotein inhibitor verapamil. Kavain concentrations were determined by reversed phase high performance liquid chromatography (HPLC).
Results
HPLC methods were developed and fully validated for kavain quantitation. Kavain demonstrated low solubility and the pH of the aqueous media did not affect its solubility. Kavain was found to be highly permeable and efflux of kavain mediated by P-glycoprotein was not significant during intestinal permeation.
Conclusion
The results of biopharmaceutical studies provided useful information for predicting availability of kavain from the gastrointestinal tract and this compound was ranked as BCS Class II, exhibiting dissolution rate-limited absorption.
da Silva Santos, Rafael Rocha; Ramos, Matheus Corrêa; Ferreira, Juliana Veloso; Gonçalves, José Eduardo; César, Isabela Costa
In: vol. 296, pp. 115480, 2022, ISSN: 1872-7573.
Abstract | Links | BibTeX | Tags: Caco-2 cell, Caco-2 Cells, Equilibrium solubility, Humans, Kava, Kavain, Lactones, permeability, Piper methysticum G. Forst, Plant Extracts, Pyrones, Solubility
@article{santos_biopharmaceutical_2022,
title = {Biopharmaceutical evaluation of kavain in Piper methysticum G. Forst dried extract: Equilibrium solubility and intestinal permeability in Caco-2 cell model},
author = {Rafael Rocha da Silva Santos and Matheus Corrêa Ramos and Juliana Veloso Ferreira and José Eduardo Gonçalves and Isabela Costa César},
doi = {10.1016/j.jep.2022.115480},
issn = {1872-7573},
year = {2022},
date = {2022-10-05},
volume = {296},
pages = {115480},
abstract = {ETHNOPHARMACOLOGICAL RELEVANCE: Piper methysticum G. Forst, popularly known as kava, is a traditional medicinal plant native from South Pacific islands and widely used to treat anxiety, depression and stress. The psychoactive properties are related to the kavalactones, mainly kavain.
AIM OF THE STUDY: To evaluate the biopharmaceutical properties of synthetic kavain and when present in kava dried extracts by means of equilibrium solubility and intestinal permeability studies in the Caco-2 cell model.
MATERIALS AND METHODS: The equilibrium solubility of kavain was performed using a shake flask incubator at 37 °C in different media at physiological pH range (1.2-6.8). The intestinal permeability of kavain evaluated in Caco-2 cells in the presence and absence of the P-glycoprotein inhibitor verapamil. Kavain concentrations were determined by reversed phase high performance liquid chromatography (HPLC).
RESULTS: HPLC methods were developed and fully validated for kavain quantitation. Kavain demonstrated low solubility and the pH of the aqueous media did not affect its solubility. Kavain was found to be highly permeable and efflux of kavain mediated by P-glycoprotein was not significant during intestinal permeation.
CONCLUSION: The results of biopharmaceutical studies provided useful information for predicting availability of kavain from the gastrointestinal tract and this compound was ranked as BCS Class II, exhibiting dissolution rate-limited absorption.},
keywords = {Caco-2 cell, Caco-2 Cells, Equilibrium solubility, Humans, Kava, Kavain, Lactones, permeability, Piper methysticum G. Forst, Plant Extracts, Pyrones, Solubility},
pubstate = {published},
tppubtype = {article}
}
ETHNOPHARMACOLOGICAL RELEVANCE: Piper methysticum G. Forst, popularly known as kava, is a traditional medicinal plant native from South Pacific islands and widely used to treat anxiety, depression and stress. The psychoactive properties are related to the kavalactones, mainly kavain.
AIM OF THE STUDY: To evaluate the biopharmaceutical properties of synthetic kavain and when present in kava dried extracts by means of equilibrium solubility and intestinal permeability studies in the Caco-2 cell model.
MATERIALS AND METHODS: The equilibrium solubility of kavain was performed using a shake flask incubator at 37 °C in different media at physiological pH range (1.2-6.8). The intestinal permeability of kavain evaluated in Caco-2 cells in the presence and absence of the P-glycoprotein inhibitor verapamil. Kavain concentrations were determined by reversed phase high performance liquid chromatography (HPLC).
RESULTS: HPLC methods were developed and fully validated for kavain quantitation. Kavain demonstrated low solubility and the pH of the aqueous media did not affect its solubility. Kavain was found to be highly permeable and efflux of kavain mediated by P-glycoprotein was not significant during intestinal permeation.
CONCLUSION: The results of biopharmaceutical studies provided useful information for predicting availability of kavain from the gastrointestinal tract and this compound was ranked as BCS Class II, exhibiting dissolution rate-limited absorption.
AIM OF THE STUDY: To evaluate the biopharmaceutical properties of synthetic kavain and when present in kava dried extracts by means of equilibrium solubility and intestinal permeability studies in the Caco-2 cell model.
MATERIALS AND METHODS: The equilibrium solubility of kavain was performed using a shake flask incubator at 37 °C in different media at physiological pH range (1.2-6.8). The intestinal permeability of kavain evaluated in Caco-2 cells in the presence and absence of the P-glycoprotein inhibitor verapamil. Kavain concentrations were determined by reversed phase high performance liquid chromatography (HPLC).
RESULTS: HPLC methods were developed and fully validated for kavain quantitation. Kavain demonstrated low solubility and the pH of the aqueous media did not affect its solubility. Kavain was found to be highly permeable and efflux of kavain mediated by P-glycoprotein was not significant during intestinal permeation.
CONCLUSION: The results of biopharmaceutical studies provided useful information for predicting availability of kavain from the gastrointestinal tract and this compound was ranked as BCS Class II, exhibiting dissolution rate-limited absorption.
Paul, Rand [R-KY Sen.
Text - S.5002 - 117th Congress (2021-2022): FDA Modernization Act 2.0 Online
2022, visited: 23.08.2024, (Archive Location: 2022-09-29).
Abstract | Links | BibTeX | Tags:
@online{sen_paul_text_2022,
title = {Text - S.5002 - 117th Congress (2021-2022): FDA Modernization Act 2.0},
author = {Rand [R-KY Sen. Paul},
url = {https://www.congress.gov/bill/117th-congress/senate-bill/5002/text},
year = {2022},
date = {2022-09-29},
urldate = {2024-08-23},
abstract = {Text for S.5002 - 117th Congress (2021-2022): FDA Modernization Act 2.0},
note = {Archive Location: 2022-09-29},
keywords = {},
pubstate = {published},
tppubtype = {online}
}
Text for S.5002 - 117th Congress (2021-2022): FDA Modernization Act 2.0
Breau, Keith A.; Ok, Meryem T.; Gomez-Martinez, Ismael; Burclaff, Joseph; Kohn, Nathan P.; Magness, Scott T.
Efficient transgenesis and homology-directed gene targeting in monolayers of primary human small intestinal and colonic epithelial stem cells Journal Article
In: vol. 17, no. 6, pp. 1493–1506, 2022, ISSN: 2213-6711.
Abstract | Links | BibTeX | Tags: {CRISPR}-Cas Systems, {CRISPR}/Cas9, {OLFM}4, {PiggyBac}, 2D monolayer cultures, electroporation, Gene Editing, Gene Targeting, human {ISC} marker, Humans, Intestine, microphysiological device, Organoids, planar crypt-microarray, Small, Stem Cells, Transfection, transgenic
@article{breau_efficient_2022,
title = {Efficient transgenesis and homology-directed gene targeting in monolayers of primary human small intestinal and colonic epithelial stem cells},
author = {Keith A. Breau and Meryem T. Ok and Ismael Gomez-Martinez and Joseph Burclaff and Nathan P. Kohn and Scott T. Magness},
doi = {10.1016/j.stemcr.2022.04.005},
issn = {2213-6711},
year = {2022},
date = {2022-06-14},
volume = {17},
number = {6},
pages = {1493–1506},
abstract = {Two-dimensional (2D) cultures of intestinal and colonic epithelium can be generated using human intestinal stem cells (hISCs) derived from primary tissue sources. These 2D cultures are emerging as attractive and versatile alternatives to three-dimensional organoid cultures; however, transgenesis and gene-editing approaches have not been developed for hISCs grown as 2D monolayers. Using 2D cultured hISCs we show that electroporation achieves up to 80% transfection in hISCs from six anatomical regions with around 64% survival and produces 0.15% transgenesis by PiggyBac transposase and 35% gene edited indels by electroporation of Cas9-ribonucleoprotein complexes at the OLFM4 locus. We create OLFM4-emGFP knock-in hISCs, validate the reporter on engineered 2D crypt devices, and develop complete workflows for high-throughput cloning and expansion of transgenic lines in 3-4 weeks. New findings demonstrate small hISCs expressing the highest OLFM4 levels exhibit the most organoid forming potential and show utility of the 2D crypt device to evaluate hISC function.},
keywords = {{CRISPR}-Cas Systems, {CRISPR}/Cas9, {OLFM}4, {PiggyBac}, 2D monolayer cultures, electroporation, Gene Editing, Gene Targeting, human {ISC} marker, Humans, Intestine, microphysiological device, Organoids, planar crypt-microarray, Small, Stem Cells, Transfection, transgenic},
pubstate = {published},
tppubtype = {article}
}
Two-dimensional (2D) cultures of intestinal and colonic epithelium can be generated using human intestinal stem cells (hISCs) derived from primary tissue sources. These 2D cultures are emerging as attractive and versatile alternatives to three-dimensional organoid cultures; however, transgenesis and gene-editing approaches have not been developed for hISCs grown as 2D monolayers. Using 2D cultured hISCs we show that electroporation achieves up to 80% transfection in hISCs from six anatomical regions with around 64% survival and produces 0.15% transgenesis by PiggyBac transposase and 35% gene edited indels by electroporation of Cas9-ribonucleoprotein complexes at the OLFM4 locus. We create OLFM4-emGFP knock-in hISCs, validate the reporter on engineered 2D crypt devices, and develop complete workflows for high-throughput cloning and expansion of transgenic lines in 3-4 weeks. New findings demonstrate small hISCs expressing the highest OLFM4 levels exhibit the most organoid forming potential and show utility of the 2D crypt device to evaluate hISC function.
Joshi, Abhishek; Soni, Arun; Acharya, Sanjeev
In vitro models and ex vivo systems used in inflammatory bowel disease Journal Article
In: vol. 1, no. 3, pp. 213–227, 2022, ISSN: 2731-3441.
Abstract | Links | BibTeX | Tags: Caco-2 cell lines, Ex vivo systems, Gut-on-chip, In vitro cell lines, inflammatory bowel disease, Organoids
@article{joshi_vitro_2022,
title = {In vitro models and ex vivo systems used in inflammatory bowel disease},
author = {Abhishek Joshi and Arun Soni and Sanjeev Acharya},
url = {https://doi.org/10.1007/s44164-022-00017-w},
doi = {10.1007/s44164-022-00017-w},
issn = {2731-3441},
year = {2022},
date = {2022-06-01},
urldate = {2023-04-07},
volume = {1},
number = {3},
pages = {213–227},
abstract = {Inflammatory bowel disease (IBD) is a chronic, relapsing gastrointestinal condition. Ulcerative colitis and Crohn’s disease are types of inflammatory bowel disease. Over many decades, the disease has been a topic of study, with experts still trying to figure out its cause and pathology. Researchers have established many in vivo animal models, in vitro cell lines, and ex vivo systems to understand its cause ultimately and adequately identify a therapy. However, in vivo animal models cannot be regarded as good models for studying IBD since they cannot completely simulate the disease. Furthermore, because species differences are a crucial subject of concern, in vitro cell lines and ex vivo systems can be employed to recreate the condition properly. In vitro models serve as the starting point for biological and medical research. Ex vivo and in vitro models for replicating gut physiology have been developed. This review aims to present a clear understanding of several in vitro and ex vivo models of IBD and provide insights into their benefits and limits and their value in understanding intestinal physiology.},
keywords = {Caco-2 cell lines, Ex vivo systems, Gut-on-chip, In vitro cell lines, inflammatory bowel disease, Organoids},
pubstate = {published},
tppubtype = {article}
}
Inflammatory bowel disease (IBD) is a chronic, relapsing gastrointestinal condition. Ulcerative colitis and Crohn’s disease are types of inflammatory bowel disease. Over many decades, the disease has been a topic of study, with experts still trying to figure out its cause and pathology. Researchers have established many in vivo animal models, in vitro cell lines, and ex vivo systems to understand its cause ultimately and adequately identify a therapy. However, in vivo animal models cannot be regarded as good models for studying IBD since they cannot completely simulate the disease. Furthermore, because species differences are a crucial subject of concern, in vitro cell lines and ex vivo systems can be employed to recreate the condition properly. In vitro models serve as the starting point for biological and medical research. Ex vivo and in vitro models for replicating gut physiology have been developed. This review aims to present a clear understanding of several in vitro and ex vivo models of IBD and provide insights into their benefits and limits and their value in understanding intestinal physiology.
Zangara, Megan T.; Ponti, András K.; Miller, Noah D.; Engelhart, Morgan J.; Ahern, Philip P.; Sangwan, Naseer; McDonald, Christine
Maltodextrin Consumption Impairs the Intestinal Mucus Barrier and Accelerates Colitis Through Direct Actions on the Epithelium Journal Article
In: vol. 13, pp. 841188, 2022, ISSN: 1664-3224.
Abstract | Links | BibTeX | Tags:
@article{zangara_maltodextrin_2022,
title = {Maltodextrin Consumption Impairs the Intestinal Mucus Barrier and Accelerates Colitis Through Direct Actions on the Epithelium},
author = {Megan T. Zangara and András K. Ponti and Noah D. Miller and Morgan J. Engelhart and Philip P. Ahern and Naseer Sangwan and Christine McDonald},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8963984/},
doi = {10.3389/fimmu.2022.841188},
issn = {1664-3224},
year = {2022},
date = {2022-03-14},
urldate = {2023-04-27},
volume = {13},
pages = {841188},
abstract = {Food additives are common components of processed foods consumed in a Western diet. In inflammatory bowel disease patients, some diets that exclude food additives improved clinical disease parameters, suggesting a link between food additives and disease pathogenesis. Food additives also enhanced disease severity in mouse colitis models through incompletely described mechanisms. This study examined the mechanisms by which the food additive maltodextrin (MDX) alters the development of colitis in a murine model. Interleukin-10 knockout (IL10KO) mice were fed diets supplemented with MDX or carboxymethyl cellulose (CMC) to determine their impact on colitis onset and severity; microbiome composition, function, and location; colonic immune cell infiltrates; and mucus layer integrity. Primary IL10KO colonic epithelial monolayers were used to dissect the impact of MDX directly on epithelial differentiation and mucus production. MDX or CMC consumption increased the incidence and severity of colitis, as well as decreased microbiome diversity, altered microbial composition, and decreased fecal acetic acid levels. The number of mucus producing cells were decreased in food additive fed mice and resulted in increased microbial proximity to the intestinal epithelium. Additionally, MDX supplementation resulted in crypt hyperplasia and expansion of the HopX+ injury renewal stem cell niche. In primary intestinal epithelial-derived monolayers devoid of microbes and immune cells, MDX exposure decreased goblet cell number and mucus production in association with downregulated expression of the transcription factor Klf4, a marker of terminally differentiated goblet cells. These results suggest MDX disrupts the balance of epithelial cell differentiation and proliferation to contribute to disease pathogenesis through direct and indirect actions on the intestinal epithelial barrier.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Food additives are common components of processed foods consumed in a Western diet. In inflammatory bowel disease patients, some diets that exclude food additives improved clinical disease parameters, suggesting a link between food additives and disease pathogenesis. Food additives also enhanced disease severity in mouse colitis models through incompletely described mechanisms. This study examined the mechanisms by which the food additive maltodextrin (MDX) alters the development of colitis in a murine model. Interleukin-10 knockout (IL10KO) mice were fed diets supplemented with MDX or carboxymethyl cellulose (CMC) to determine their impact on colitis onset and severity; microbiome composition, function, and location; colonic immune cell infiltrates; and mucus layer integrity. Primary IL10KO colonic epithelial monolayers were used to dissect the impact of MDX directly on epithelial differentiation and mucus production. MDX or CMC consumption increased the incidence and severity of colitis, as well as decreased microbiome diversity, altered microbial composition, and decreased fecal acetic acid levels. The number of mucus producing cells were decreased in food additive fed mice and resulted in increased microbial proximity to the intestinal epithelium. Additionally, MDX supplementation resulted in crypt hyperplasia and expansion of the HopX+ injury renewal stem cell niche. In primary intestinal epithelial-derived monolayers devoid of microbes and immune cells, MDX exposure decreased goblet cell number and mucus production in association with downregulated expression of the transcription factor Klf4, a marker of terminally differentiated goblet cells. These results suggest MDX disrupts the balance of epithelial cell differentiation and proliferation to contribute to disease pathogenesis through direct and indirect actions on the intestinal epithelial barrier.
Burclaff, Joseph; Bliton, R. Jarrett; Breau, Keith A.; Ok, Meryem T.; Gomez-Martinez, Ismael; Ranek, Jolene S.; Bhatt, Aadra P.; Purvis, Jeremy E.; Woosley, John T.; Magness, Scott T.
A Proximal-to-Distal Survey of Healthy Adult Human Small Intestine and Colon Epithelium by Single-Cell Transcriptomics Journal Article
In: vol. 13, no. 5, pp. 1554–1589, 2022, ISSN: 2352-345X.
Abstract | Links | BibTeX | Tags:
@article{burclaff_proximal–distal_2022,
title = {A Proximal-to-Distal Survey of Healthy Adult Human Small Intestine and Colon Epithelium by Single-Cell Transcriptomics},
author = {Joseph Burclaff and R. Jarrett Bliton and Keith A. Breau and Meryem T. Ok and Ismael Gomez-Martinez and Jolene S. Ranek and Aadra P. Bhatt and Jeremy E. Purvis and John T. Woosley and Scott T. Magness},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9043569/},
doi = {10.1016/j.jcmgh.2022.02.007},
issn = {2352-345X},
year = {2022},
date = {2022-02-15},
urldate = {2024-07-31},
volume = {13},
number = {5},
pages = {1554–1589},
abstract = {Background & Aims
Single-cell transcriptomics offer unprecedented resolution of tissue function at the cellular level, yet studies analyzing healthy adult human small intestine and colon are sparse. Here, we present single-cell transcriptomics covering the duodenum, jejunum, ileum, and ascending, transverse, and descending colon from 3 human beings.
Methods
A total of 12,590 single epithelial cells from 3 independently processed organ donors were evaluated for organ-specific lineage biomarkers, differentially regulated genes, receptors, and drug targets. Analyses focused on intrinsic cell properties and their capacity for response to extrinsic signals along the gut axis across different human beings.
Results
Cells were assigned to 25 epithelial lineage clusters. Multiple accepted intestinal stem cell markers do not specifically mark all human intestinal stem cells. Lysozyme expression is not unique to human Paneth cells, and Paneth cells lack expression of expected niche factors. Bestrophin 4 (BEST4)+ cells express Neuropeptide Y (NPY) and show maturational differences between the small intestine and colon. Tuft cells possess a broad ability to interact with the innate and adaptive immune systems through previously unreported receptors. Some classes of mucins, hormones, cell junctions, and nutrient absorption genes show unappreciated regional expression differences across lineages. The differential expression of receptors and drug targets across lineages show biological variation and the potential for variegated responses.
Conclusions
Our study identifies novel lineage marker genes, covers regional differences, shows important differences between mouse and human gut epithelium, and reveals insight into how the epithelium responds to the environment and drugs. This comprehensive cell atlas of the healthy adult human intestinal epithelium resolves likely functional differences across anatomic regions along the gastrointestinal tract and advances our understanding of human intestinal physiology.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Background & Aims
Single-cell transcriptomics offer unprecedented resolution of tissue function at the cellular level, yet studies analyzing healthy adult human small intestine and colon are sparse. Here, we present single-cell transcriptomics covering the duodenum, jejunum, ileum, and ascending, transverse, and descending colon from 3 human beings.
Methods
A total of 12,590 single epithelial cells from 3 independently processed organ donors were evaluated for organ-specific lineage biomarkers, differentially regulated genes, receptors, and drug targets. Analyses focused on intrinsic cell properties and their capacity for response to extrinsic signals along the gut axis across different human beings.
Results
Cells were assigned to 25 epithelial lineage clusters. Multiple accepted intestinal stem cell markers do not specifically mark all human intestinal stem cells. Lysozyme expression is not unique to human Paneth cells, and Paneth cells lack expression of expected niche factors. Bestrophin 4 (BEST4)+ cells express Neuropeptide Y (NPY) and show maturational differences between the small intestine and colon. Tuft cells possess a broad ability to interact with the innate and adaptive immune systems through previously unreported receptors. Some classes of mucins, hormones, cell junctions, and nutrient absorption genes show unappreciated regional expression differences across lineages. The differential expression of receptors and drug targets across lineages show biological variation and the potential for variegated responses.
Conclusions
Our study identifies novel lineage marker genes, covers regional differences, shows important differences between mouse and human gut epithelium, and reveals insight into how the epithelium responds to the environment and drugs. This comprehensive cell atlas of the healthy adult human intestinal epithelium resolves likely functional differences across anatomic regions along the gastrointestinal tract and advances our understanding of human intestinal physiology.
Single-cell transcriptomics offer unprecedented resolution of tissue function at the cellular level, yet studies analyzing healthy adult human small intestine and colon are sparse. Here, we present single-cell transcriptomics covering the duodenum, jejunum, ileum, and ascending, transverse, and descending colon from 3 human beings.
Methods
A total of 12,590 single epithelial cells from 3 independently processed organ donors were evaluated for organ-specific lineage biomarkers, differentially regulated genes, receptors, and drug targets. Analyses focused on intrinsic cell properties and their capacity for response to extrinsic signals along the gut axis across different human beings.
Results
Cells were assigned to 25 epithelial lineage clusters. Multiple accepted intestinal stem cell markers do not specifically mark all human intestinal stem cells. Lysozyme expression is not unique to human Paneth cells, and Paneth cells lack expression of expected niche factors. Bestrophin 4 (BEST4)+ cells express Neuropeptide Y (NPY) and show maturational differences between the small intestine and colon. Tuft cells possess a broad ability to interact with the innate and adaptive immune systems through previously unreported receptors. Some classes of mucins, hormones, cell junctions, and nutrient absorption genes show unappreciated regional expression differences across lineages. The differential expression of receptors and drug targets across lineages show biological variation and the potential for variegated responses.
Conclusions
Our study identifies novel lineage marker genes, covers regional differences, shows important differences between mouse and human gut epithelium, and reveals insight into how the epithelium responds to the environment and drugs. This comprehensive cell atlas of the healthy adult human intestinal epithelium resolves likely functional differences across anatomic regions along the gastrointestinal tract and advances our understanding of human intestinal physiology.
Arian, Christopher M.; Imaoka, Tomoki; Yang, Jade; Kelly, Edward J.; Thummel, Kenneth E.
Gutsy science: In vitro systems of the human intestine to model oral drug disposition Journal Article
In: vol. 230, pp. 107962, 2022, ISSN: 0163-7258.
Abstract | Links | BibTeX | Tags: first-pass, Intestine, Microphysiological systems, Organoids
@article{arian_gutsy_2022,
title = {Gutsy science: In vitro systems of the human intestine to model oral drug disposition},
author = {Christopher M. Arian and Tomoki Imaoka and Jade Yang and Edward J. Kelly and Kenneth E. Thummel},
url = {https://www.sciencedirect.com/science/article/pii/S0163725821001649},
doi = {10.1016/j.pharmthera.2021.107962},
issn = {0163-7258},
year = {2022},
date = {2022-02-01},
urldate = {2023-06-16},
volume = {230},
pages = {107962},
abstract = {The intestine has important gate-keeping functions that can profoundly affect the systemic blood exposure of orally administered drugs. Thus, characterizing a new molecular entity's (NME) disposition within the intestine is of utmost importance in drug development. While currently used in vitro systems, such as Ussing chamber, precision-cut intestinal slices, immortalized cell lines, and primary enterocytes provide substantial knowledge about drug absorption and the intestinal first-pass effect, they remain sub-optimal for quantitatively predicting this process and the oral bioavailability of many drugs. Use of novel in vitro systems such as intestinal organoids and intestinal microphysiological systems have provided substantial advances over the past decade, expanding our understanding of intestinal physiology, pathology, and development. However, application of these emerging in vitro systems in the pharmaceutical science is in its infancy. Preliminary work has demonstrated that these systems more accurately recapitulate the physiology and biochemistry of the intact intestine, as it relates to oral drug disposition, and thus they hold considerable promise as preclinical testing platforms of the future. Here we review currently used and emerging in vitro models of the human intestine employed in pharmaceutical science research. We also highlight aspects of these emerging tools that require further study.},
keywords = {first-pass, Intestine, Microphysiological systems, Organoids},
pubstate = {published},
tppubtype = {article}
}
The intestine has important gate-keeping functions that can profoundly affect the systemic blood exposure of orally administered drugs. Thus, characterizing a new molecular entity's (NME) disposition within the intestine is of utmost importance in drug development. While currently used in vitro systems, such as Ussing chamber, precision-cut intestinal slices, immortalized cell lines, and primary enterocytes provide substantial knowledge about drug absorption and the intestinal first-pass effect, they remain sub-optimal for quantitatively predicting this process and the oral bioavailability of many drugs. Use of novel in vitro systems such as intestinal organoids and intestinal microphysiological systems have provided substantial advances over the past decade, expanding our understanding of intestinal physiology, pathology, and development. However, application of these emerging in vitro systems in the pharmaceutical science is in its infancy. Preliminary work has demonstrated that these systems more accurately recapitulate the physiology and biochemistry of the intact intestine, as it relates to oral drug disposition, and thus they hold considerable promise as preclinical testing platforms of the future. Here we review currently used and emerging in vitro models of the human intestine employed in pharmaceutical science research. We also highlight aspects of these emerging tools that require further study.
Macedo, Maria Helena; Barros, Andreia S.; Martínez, Elena; Barrias, Cristina C.; Sarmento, Bruno
All layers matter: Innovative three-dimensional epithelium-stroma-endothelium intestinal model for reliable permeability outcomes Journal Article
In: vol. 341, pp. 414–430, 2022, ISSN: 0168-3659.
Abstract | Links | BibTeX | Tags: 3D intestinal model, drug absorption, Drug development, Endothelium, Hydrogel, permeability
@article{macedo_all_2022,
title = {All layers matter: Innovative three-dimensional epithelium-stroma-endothelium intestinal model for reliable permeability outcomes},
author = {Maria Helena Macedo and Andreia S. Barros and Elena Martínez and Cristina C. Barrias and Bruno Sarmento},
url = {https://www.sciencedirect.com/science/article/pii/S0168365921006519},
doi = {10.1016/j.jconrel.2021.11.048},
issn = {0168-3659},
year = {2022},
date = {2022-01-01},
urldate = {2024-04-02},
volume = {341},
pages = {414–430},
abstract = {Drug development is an ever-growing field, increasingly requesting reliable in vitro tools to speed up early screening phases, reducing the need for animal experiments. In oral delivery, understanding the absorption pattern of a new drug in the small intestine is paramount. Classical two-dimensional (2D) in vitro models are generally too simplistic and do not accurately represent native tissues. The main goal of this work was to develop an advanced three-dimensional (3D) in vitro intestinal model to test absorption in a more reliable manner, by better mimicking the native environment. The 3D model is composed of a collagen-based stromal layer with embedded fibroblasts mimicking the intestinal lamina propria and providing support for the epithelium, composed of enterocytes and mucus-secreting cells. An endothelial layer, surrogating the absorptive capillary network, is also present. The cellular crosstalk between the different cells present in the model is unveiled, disclosing key players, namely those involved in the contraction of collagen by fibroblasts. The developed 3D model presents lower levels of P-glycoprotein (P-gp) and Multidrug Resistance Protein 2 (MRP2) efflux transporters, which are normally overexpressed in traditional Caco-2 models, and are paramount in the absorption of many compounds. This, allied with transepithelial electrical resistance (TEER) values closer to physiological ranges, leads to improved and more reliable permeability outcomes, which are observed when comparing our results with in vivo data.},
keywords = {3D intestinal model, drug absorption, Drug development, Endothelium, Hydrogel, permeability},
pubstate = {published},
tppubtype = {article}
}
Drug development is an ever-growing field, increasingly requesting reliable in vitro tools to speed up early screening phases, reducing the need for animal experiments. In oral delivery, understanding the absorption pattern of a new drug in the small intestine is paramount. Classical two-dimensional (2D) in vitro models are generally too simplistic and do not accurately represent native tissues. The main goal of this work was to develop an advanced three-dimensional (3D) in vitro intestinal model to test absorption in a more reliable manner, by better mimicking the native environment. The 3D model is composed of a collagen-based stromal layer with embedded fibroblasts mimicking the intestinal lamina propria and providing support for the epithelium, composed of enterocytes and mucus-secreting cells. An endothelial layer, surrogating the absorptive capillary network, is also present. The cellular crosstalk between the different cells present in the model is unveiled, disclosing key players, namely those involved in the contraction of collagen by fibroblasts. The developed 3D model presents lower levels of P-glycoprotein (P-gp) and Multidrug Resistance Protein 2 (MRP2) efflux transporters, which are normally overexpressed in traditional Caco-2 models, and are paramount in the absorption of many compounds. This, allied with transepithelial electrical resistance (TEER) values closer to physiological ranges, leads to improved and more reliable permeability outcomes, which are observed when comparing our results with in vivo data.
2021
Elzinga, Janneke; Lugt, Benthe; Belzer, Clara; Steegenga, Wilma T.
Characterization of increased mucus production of HT29-MTX-E12 cells grown under Semi-Wet interface with Mechanical Stimulation Journal Article
In: vol. 16, no. 12, pp. e0261191, 2021, ISSN: 1932-6203.
Abstract | Links | BibTeX | Tags: Caco-2 Cells, Cell differentiation, Gastrointestinal tract, Gene Expression, Glucose, Microarrays, Mucin, Mucus
@article{elzinga_characterization_2021,
title = {Characterization of increased mucus production of HT29-MTX-E12 cells grown under Semi-Wet interface with Mechanical Stimulation},
author = {Janneke Elzinga and Benthe Lugt and Clara Belzer and Wilma T. Steegenga},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8687553/},
doi = {10.1371/journal.pone.0261191},
issn = {1932-6203},
year = {2021},
date = {2021-12-20},
urldate = {2024-04-02},
volume = {16},
number = {12},
pages = {e0261191},
abstract = {The intestinal mucus layer plays a crucial role in human health. To study intestinal mucus function and structure in vitro, the mucus-producing intestinal cell line HT29-MTX-E12 has been commonly used. However, this cell line produces only low amounts of the intestine-specific MUC2. It has been shown previously that HT29-MTX-E12 cells cultured under Semi-Wet interface with Mechanical Stimulation (SWMS) produced higher amounts of MUC2, concomitant with a thicker mucus layer, compared to cells cultured conventionally. However, it remains unknown which underlying pathways are involved. Therefore, we aimed to further explore the cellular processes underlying the increased MUC2 production by HT29-MTX-E12 cells grown under SWMS conditions. Cells grown on Transwell membranes for 14 days under static and SWMS conditions (after cell seeding and attachment) were subjected to transcriptome analysis to investigate underlying molecular pathways at gene expression level. Caco-2 and LS174T cell lines were included as references. We characterized how SWMS conditions affected HT29-MTX-E12 cells in terms of epithelial barrier integrity, by measuring transepithelial electrical resistance, and cell metabolism, by monitoring pH and lactate production per molecule glucose of the conditioned medium. We confirmed higher MUC2 production under SWMS conditions at gene and protein level and demonstrated that this culturing method primarily stimulated cell growth. In addition, we also found evidence for a more aerobic cell metabolism under SWMS, as shown previously for similar models. In summary, we suggest different mechanisms by which MUC2 production is enhanced under SWMS and propose potential applications of this model in future studies.},
keywords = {Caco-2 Cells, Cell differentiation, Gastrointestinal tract, Gene Expression, Glucose, Microarrays, Mucin, Mucus},
pubstate = {published},
tppubtype = {article}
}
The intestinal mucus layer plays a crucial role in human health. To study intestinal mucus function and structure in vitro, the mucus-producing intestinal cell line HT29-MTX-E12 has been commonly used. However, this cell line produces only low amounts of the intestine-specific MUC2. It has been shown previously that HT29-MTX-E12 cells cultured under Semi-Wet interface with Mechanical Stimulation (SWMS) produced higher amounts of MUC2, concomitant with a thicker mucus layer, compared to cells cultured conventionally. However, it remains unknown which underlying pathways are involved. Therefore, we aimed to further explore the cellular processes underlying the increased MUC2 production by HT29-MTX-E12 cells grown under SWMS conditions. Cells grown on Transwell membranes for 14 days under static and SWMS conditions (after cell seeding and attachment) were subjected to transcriptome analysis to investigate underlying molecular pathways at gene expression level. Caco-2 and LS174T cell lines were included as references. We characterized how SWMS conditions affected HT29-MTX-E12 cells in terms of epithelial barrier integrity, by measuring transepithelial electrical resistance, and cell metabolism, by monitoring pH and lactate production per molecule glucose of the conditioned medium. We confirmed higher MUC2 production under SWMS conditions at gene and protein level and demonstrated that this culturing method primarily stimulated cell growth. In addition, we also found evidence for a more aerobic cell metabolism under SWMS, as shown previously for similar models. In summary, we suggest different mechanisms by which MUC2 production is enhanced under SWMS and propose potential applications of this model in future studies.
Baydi, Zineb; Limami, Youness; Khalki, Loubna; Zaid, Nabil; Naya, Abdallah; Mtairag, El Mostafa; Oudghiri, Mounia; Zaid, Younes
An Update of Research Animal Models of Inflammatory Bowel Disease Journal Article
In: vol. 2021, pp. 7479540, 2021, ISSN: 2356-6140.
Abstract | Links | BibTeX | Tags:
@article{baydi_update_2021,
title = {An Update of Research Animal Models of Inflammatory Bowel Disease},
author = {Zineb Baydi and Youness Limami and Loubna Khalki and Nabil Zaid and Abdallah Naya and El Mostafa Mtairag and Mounia Oudghiri and Younes Zaid},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8687830/},
doi = {10.1155/2021/7479540},
issn = {2356-6140},
year = {2021},
date = {2021-12-13},
urldate = {2023-02-02},
volume = {2021},
pages = {7479540},
abstract = {Inflammatory bowel disease (IBD) is a group of chronic disorders that includes two main disease forms, Crohn's disease, and ulcerative colitis. The understanding of the intestinal inflammation occurring in IBD has been immeasurably advanced by the development of the now numerous murine models of intestinal inflammation. The usefulness of this research tool in IBD arises from a convergence of underlying genetic susceptibility, immune system dysfunction, environmental factors, and shifts in gut microbiota. Due to the multifactorial feature of these diseases, different animal models have been used to investigate the underlying mechanisms and develop potential therapeutic strategies. The results of preclinical efficacy studies often inform the progression of therapeutic strategies. This review describes the distinct feature and limitations of each murine IBD model and discusses the previous and current lessons from the IBD models.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Inflammatory bowel disease (IBD) is a group of chronic disorders that includes two main disease forms, Crohn's disease, and ulcerative colitis. The understanding of the intestinal inflammation occurring in IBD has been immeasurably advanced by the development of the now numerous murine models of intestinal inflammation. The usefulness of this research tool in IBD arises from a convergence of underlying genetic susceptibility, immune system dysfunction, environmental factors, and shifts in gut microbiota. Due to the multifactorial feature of these diseases, different animal models have been used to investigate the underlying mechanisms and develop potential therapeutic strategies. The results of preclinical efficacy studies often inform the progression of therapeutic strategies. This review describes the distinct feature and limitations of each murine IBD model and discusses the previous and current lessons from the IBD models.
Howard, R. Logan; Markovetz, Matthew; Wang, Yuli; Ehre, Camille; Sheikh, Shehzad Z.; Allbritton, Nancy L.; Hill, David B.
Biochemical and rheological analysis of human colonic culture mucus reveals similarity to gut mucus Journal Article
In: vol. 120, no. 23, pp. 5384–5394, 2021, ISSN: 0006-3495, (Publisher: Elsevier).
@article{howard_biochemical_2021,
title = {Biochemical and rheological analysis of human colonic culture mucus reveals similarity to gut mucus},
author = {R. Logan Howard and Matthew Markovetz and Yuli Wang and Camille Ehre and Shehzad Z. Sheikh and Nancy L. Allbritton and David B. Hill},
url = {https://www.cell.com/biophysj/abstract/S0006-3495(21)00883-3},
doi = {10.1016/j.bpj.2021.10.024},
issn = {0006-3495},
year = {2021},
date = {2021-12-07},
urldate = {2023-07-03},
volume = {120},
number = {23},
pages = {5384–5394},
note = {Publisher: Elsevier},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Franco, Yesenia L.; Silva, Lais Da; Cristofoletti, Rodrigo
Navigating Through Cell-Based In vitro Models Available for Prediction of Intestinal Permeability and Metabolism: Are We Ready for 3D? Journal Article
In: vol. 24, no. 1, pp. 2, 2021, ISSN: 1550-7416.
Abstract | Links | BibTeX | Tags: cell culture, in vitro models, microdevices, microphysiological system, Organoids, permeability
@article{franco_navigating_2021,
title = {Navigating Through Cell-Based In vitro Models Available for Prediction of Intestinal Permeability and Metabolism: Are We Ready for 3D?},
author = {Yesenia L. Franco and Lais Da Silva and Rodrigo Cristofoletti},
url = {https://doi.org/10.1208/s12248-021-00665-y},
doi = {10.1208/s12248-021-00665-y},
issn = {1550-7416},
year = {2021},
date = {2021-11-22},
urldate = {2022-12-01},
volume = {24},
number = {1},
pages = {2},
abstract = {Traditionally, in vitro studies to quantify theintestinal permeability of drugs have relied on two-dimensional cell culture models using human colorectal carcinoma cell lines, namely Caco-2, HT 29 and T84 cells. Although these models have been commonly used for high-throughput screening of xenobiotics in preclinical studies, they do not fully recapitulate the morphology and functionality of enterocytes found in the human intestine in vivo. Efforts to improve the physiological and functional relevance of in vitro intestinal models have led to the development of enteroids/intestinal organoids and microphysiological systems. These models leverage advances in three-dimensional cell culture techniques and stem cell technology (in addition to microfluidics for microphysiological systems), to mimic the architecture and microenvironment of the in vivo intestine more accurately. In this commentary, we will discuss the advantages and limitations of these established and emerging intestinal models, as well as their current and potential future applications for the pre-clinical assessment of oral therapies.},
keywords = {cell culture, in vitro models, microdevices, microphysiological system, Organoids, permeability},
pubstate = {published},
tppubtype = {article}
}
Traditionally, in vitro studies to quantify theintestinal permeability of drugs have relied on two-dimensional cell culture models using human colorectal carcinoma cell lines, namely Caco-2, HT 29 and T84 cells. Although these models have been commonly used for high-throughput screening of xenobiotics in preclinical studies, they do not fully recapitulate the morphology and functionality of enterocytes found in the human intestine in vivo. Efforts to improve the physiological and functional relevance of in vitro intestinal models have led to the development of enteroids/intestinal organoids and microphysiological systems. These models leverage advances in three-dimensional cell culture techniques and stem cell technology (in addition to microfluidics for microphysiological systems), to mimic the architecture and microenvironment of the in vivo intestine more accurately. In this commentary, we will discuss the advantages and limitations of these established and emerging intestinal models, as well as their current and potential future applications for the pre-clinical assessment of oral therapies.
Honig, Gerard; Larkin, Paul B; Heller, Caren; Hurtado-Lorenzo, Andrés
Research-Based Product Innovation to Address Critical Unmet Needs of Patients with Inflammatory Bowel Diseases Journal Article
In: vol. 27, iss. Suppl 2, pp. S1–S16, 2021, ISSN: 1078-0998.
Abstract | Links | BibTeX | Tags:
@article{honig_research-based_2021,
title = {Research-Based Product Innovation to Address Critical Unmet Needs of Patients with Inflammatory Bowel Diseases},
author = {Gerard Honig and Paul B Larkin and Caren Heller and Andrés Hurtado-Lorenzo},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8922161/},
doi = {10.1093/ibd/izab230},
issn = {1078-0998},
year = {2021},
date = {2021-11-15},
urldate = {2023-04-27},
volume = {27},
issue = {Suppl 2},
pages = {S1–S16},
abstract = {Despite progress in recent decades, patients with inflammatory bowel diseases face many
critical unmet needs, demonstrating the limitations of available treatment options.
Addressing these unmet needs will require interventions targeting multiple aspects of
inflammatory bowel disease pathology, including disease drivers that are not targeted by
available therapies. The vast majority of late-stage investigational therapies also focus
primarily on a narrow range of fundamental mechanisms. Thus, there is a pressing need to
advance to clinical stage differentiated investigational therapies directly targeting a
broader range of key mechanistic drivers of inflammatory bowel diseases. In addition,
innovations are critically needed to enable treatments to be tailored to the specific
underlying abnormal biological pathways of patients; interventions with improved safety
profiles; biomarkers to develop prognostic, predictive, and monitoring tests; novel
devices for nonpharmacological approaches such as minimally invasive monitoring; and
digital health technologies. To address these needs, the Crohn’s & Colitis Foundation
launched IBD Ventures, a venture philanthropy–funding mechanism, and IBD Innovate®, an
innovative, product-focused scientific conference. This special IBD Innovate® supplement
is a collection of articles reflecting the diverse and exciting research and development
that is currently ongoing in the inflammatory bowel disease field to deliver innovative
and differentiated products addressing critical unmet needs of patients. Here, we
highlight the pipeline of new product opportunities currently advancing at the preclinical
and early clinical development stages. We categorize and describe novel and differentiated
potential product opportunities based on their potential to address the following critical
unmet patient needs: (1) biomarkers for prognosis of disease course and
prediction/monitoring of treatment response; (2) restoration of eubiosis; (3) restoration
of barrier function and mucosal healing; (4) more effective and safer anti-inflammatories;
(5) neuromodulatory and behavioral therapies; (6) management of disease complications; and
(7) targeted drug delivery., We highlight the pipeline of investigational therapies, diagnostics, and devices with
potential to address pressing unmet needs of patients with inflammatory bowel diseases,
including biomarkers for prognosis and treatment response, restoration of eubiosis and
mucosal healing, neuromodulation, and improved control of inflammation and disease
complications.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Despite progress in recent decades, patients with inflammatory bowel diseases face many
critical unmet needs, demonstrating the limitations of available treatment options.
Addressing these unmet needs will require interventions targeting multiple aspects of
inflammatory bowel disease pathology, including disease drivers that are not targeted by
available therapies. The vast majority of late-stage investigational therapies also focus
primarily on a narrow range of fundamental mechanisms. Thus, there is a pressing need to
advance to clinical stage differentiated investigational therapies directly targeting a
broader range of key mechanistic drivers of inflammatory bowel diseases. In addition,
innovations are critically needed to enable treatments to be tailored to the specific
underlying abnormal biological pathways of patients; interventions with improved safety
profiles; biomarkers to develop prognostic, predictive, and monitoring tests; novel
devices for nonpharmacological approaches such as minimally invasive monitoring; and
digital health technologies. To address these needs, the Crohn’s & Colitis Foundation
launched IBD Ventures, a venture philanthropy–funding mechanism, and IBD Innovate®, an
innovative, product-focused scientific conference. This special IBD Innovate® supplement
is a collection of articles reflecting the diverse and exciting research and development
that is currently ongoing in the inflammatory bowel disease field to deliver innovative
and differentiated products addressing critical unmet needs of patients. Here, we
highlight the pipeline of new product opportunities currently advancing at the preclinical
and early clinical development stages. We categorize and describe novel and differentiated
potential product opportunities based on their potential to address the following critical
unmet patient needs: (1) biomarkers for prognosis of disease course and
prediction/monitoring of treatment response; (2) restoration of eubiosis; (3) restoration
of barrier function and mucosal healing; (4) more effective and safer anti-inflammatories;
(5) neuromodulatory and behavioral therapies; (6) management of disease complications; and
(7) targeted drug delivery., We highlight the pipeline of investigational therapies, diagnostics, and devices with
potential to address pressing unmet needs of patients with inflammatory bowel diseases,
including biomarkers for prognosis and treatment response, restoration of eubiosis and
mucosal healing, neuromodulation, and improved control of inflammation and disease
complications.
critical unmet needs, demonstrating the limitations of available treatment options.
Addressing these unmet needs will require interventions targeting multiple aspects of
inflammatory bowel disease pathology, including disease drivers that are not targeted by
available therapies. The vast majority of late-stage investigational therapies also focus
primarily on a narrow range of fundamental mechanisms. Thus, there is a pressing need to
advance to clinical stage differentiated investigational therapies directly targeting a
broader range of key mechanistic drivers of inflammatory bowel diseases. In addition,
innovations are critically needed to enable treatments to be tailored to the specific
underlying abnormal biological pathways of patients; interventions with improved safety
profiles; biomarkers to develop prognostic, predictive, and monitoring tests; novel
devices for nonpharmacological approaches such as minimally invasive monitoring; and
digital health technologies. To address these needs, the Crohn’s & Colitis Foundation
launched IBD Ventures, a venture philanthropy–funding mechanism, and IBD Innovate®, an
innovative, product-focused scientific conference. This special IBD Innovate® supplement
is a collection of articles reflecting the diverse and exciting research and development
that is currently ongoing in the inflammatory bowel disease field to deliver innovative
and differentiated products addressing critical unmet needs of patients. Here, we
highlight the pipeline of new product opportunities currently advancing at the preclinical
and early clinical development stages. We categorize and describe novel and differentiated
potential product opportunities based on their potential to address the following critical
unmet patient needs: (1) biomarkers for prognosis of disease course and
prediction/monitoring of treatment response; (2) restoration of eubiosis; (3) restoration
of barrier function and mucosal healing; (4) more effective and safer anti-inflammatories;
(5) neuromodulatory and behavioral therapies; (6) management of disease complications; and
(7) targeted drug delivery., We highlight the pipeline of investigational therapies, diagnostics, and devices with
potential to address pressing unmet needs of patients with inflammatory bowel diseases,
including biomarkers for prognosis and treatment response, restoration of eubiosis and
mucosal healing, neuromodulation, and improved control of inflammation and disease
complications.
Mohammadi, Sina; Morell-Perez, Carolina; Wright, Charles W.; Wyche, Thomas P.; White, Cory H.; Sana, Theodore R.; Lieberman, Linda A.
Assessing donor-to-donor variability in human intestinal organoid cultures Journal Article
In: vol. 16, no. 9, pp. 2364–2378, 2021, ISSN: 2213-6711.
Abstract | Links | BibTeX | Tags: hormones, human, Intestine, metabolism, Organoids, variability
@article{mohammadi_assessing_2021,
title = {Assessing donor-to-donor variability in human intestinal organoid cultures},
author = {Sina Mohammadi and Carolina Morell-Perez and Charles W. Wright and Thomas P. Wyche and Cory H. White and Theodore R. Sana and Linda A. Lieberman},
url = {https://www.sciencedirect.com/science/article/pii/S2213671121003829},
doi = {10.1016/j.stemcr.2021.07.016},
issn = {2213-6711},
year = {2021},
date = {2021-09-14},
urldate = {2022-11-15},
volume = {16},
number = {9},
pages = {2364–2378},
abstract = {Donor-to-donor variability in primary human organoid cultures has not been well characterized. As these cultures contain multiple cell types, there is greater concern that variability could lead to increased noise. In this work we investigated donor-to-donor variability in human gut adult stem cell (ASC) organoids. We examined intestinal developmental pathways during culture differentiation in ileum- and colon-derived cultures established from multiple donors, showing that differentiation patterns were consistent among cultures. This finding indicates that donor-to-donor variability in this system remains at a manageable level. Intestinal metabolic activity was evaluated by targeted analysis of central carbon metabolites and by analyzing hormone production patterns. Both experiments demonstrated similar metabolic functions among donors. Importantly, this activity reflected intestinal biology, indicating that these ASC organoid cultures are appropriate for studying metabolic processes. This work establishes a framework for generating high-confidence data using human primary cultures through thorough characterization of variability.},
keywords = {hormones, human, Intestine, metabolism, Organoids, variability},
pubstate = {published},
tppubtype = {article}
}
Donor-to-donor variability in primary human organoid cultures has not been well characterized. As these cultures contain multiple cell types, there is greater concern that variability could lead to increased noise. In this work we investigated donor-to-donor variability in human gut adult stem cell (ASC) organoids. We examined intestinal developmental pathways during culture differentiation in ileum- and colon-derived cultures established from multiple donors, showing that differentiation patterns were consistent among cultures. This finding indicates that donor-to-donor variability in this system remains at a manageable level. Intestinal metabolic activity was evaluated by targeted analysis of central carbon metabolites and by analyzing hormone production patterns. Both experiments demonstrated similar metabolic functions among donors. Importantly, this activity reflected intestinal biology, indicating that these ASC organoid cultures are appropriate for studying metabolic processes. This work establishes a framework for generating high-confidence data using human primary cultures through thorough characterization of variability.
Vinarov, Zahari; Abdallah, Mohammad; Agundez, José A. G.; Allegaert, Karel; Basit, Abdul W.; Braeckmans, Marlies; Ceulemans, Jens; Corsetti, Maura; Griffin, Brendan T.; Grimm, Michael; Keszthelyi, Daniel; Koziolek, Mirko; Madla, Christine M.; Matthys, Christophe; McCoubrey, Laura E.; Mitra, Amitava; Reppas, Christos; Stappaerts, Jef; Steenackers, Nele; Trevaskis, Natalie L.; Vanuytsel, Tim; Vertzoni, Maria; Weitschies, Werner; Wilson, Clive; Augustijns, Patrick
Impact of gastrointestinal tract variability on oral drug absorption and pharmacokinetics: An UNGAP review Journal Article
In: vol. 162, pp. 105812, 2021, ISSN: 0928-0987.
Abstract | Links | BibTeX | Tags: Diseases, Drug formulation, Fasted and fed state, Pediatrics and geriatrics, Physiology, Variation
@article{vinarov_impact_2021,
title = {Impact of gastrointestinal tract variability on oral drug absorption and pharmacokinetics: An UNGAP review},
author = {Zahari Vinarov and Mohammad Abdallah and José A. G. Agundez and Karel Allegaert and Abdul W. Basit and Marlies Braeckmans and Jens Ceulemans and Maura Corsetti and Brendan T. Griffin and Michael Grimm and Daniel Keszthelyi and Mirko Koziolek and Christine M. Madla and Christophe Matthys and Laura E. McCoubrey and Amitava Mitra and Christos Reppas and Jef Stappaerts and Nele Steenackers and Natalie L. Trevaskis and Tim Vanuytsel and Maria Vertzoni and Werner Weitschies and Clive Wilson and Patrick Augustijns},
url = {https://www.sciencedirect.com/science/article/pii/S0928098721001147},
doi = {10.1016/j.ejps.2021.105812},
issn = {0928-0987},
year = {2021},
date = {2021-07-01},
urldate = {2025-02-28},
volume = {162},
pages = {105812},
abstract = {The absorption of oral drugs is frequently plagued by significant variability with potentially serious therapeutic consequences. The source of variability can be traced back to interindividual variability in physiology, differences in special populations (age- and disease-dependent), drug and formulation properties, or food-drug interactions. Clinical evidence for the impact of some of these factors on drug pharmacokinetic variability is mounting: e.g. gastric pH and emptying time, small intestinal fluid properties, differences in pediatrics and the elderly, and surgical changes in gastrointestinal anatomy. However, the link of colonic factors variability (transit time, fluid composition, microbiome), sex differences (male vs. female) and gut-related diseases (chronic constipation, anorexia and cachexia) to drug absorption variability has not been firmly established yet. At the same time, a way to decrease oral drug pharmacokinetic variability is provided by the pharmaceutical industry: clinical evidence suggests that formulation approaches employed during drug development can decrease the variability in oral exposure. This review outlines the main drivers of oral drug exposure variability and potential approaches to overcome them, while highlighting existing knowledge gaps and guiding future studies in this area.},
keywords = {Diseases, Drug formulation, Fasted and fed state, Pediatrics and geriatrics, Physiology, Variation},
pubstate = {published},
tppubtype = {article}
}
The absorption of oral drugs is frequently plagued by significant variability with potentially serious therapeutic consequences. The source of variability can be traced back to interindividual variability in physiology, differences in special populations (age- and disease-dependent), drug and formulation properties, or food-drug interactions. Clinical evidence for the impact of some of these factors on drug pharmacokinetic variability is mounting: e.g. gastric pH and emptying time, small intestinal fluid properties, differences in pediatrics and the elderly, and surgical changes in gastrointestinal anatomy. However, the link of colonic factors variability (transit time, fluid composition, microbiome), sex differences (male vs. female) and gut-related diseases (chronic constipation, anorexia and cachexia) to drug absorption variability has not been firmly established yet. At the same time, a way to decrease oral drug pharmacokinetic variability is provided by the pharmaceutical industry: clinical evidence suggests that formulation approaches employed during drug development can decrease the variability in oral exposure. This review outlines the main drivers of oral drug exposure variability and potential approaches to overcome them, while highlighting existing knowledge gaps and guiding future studies in this area.
Koepsell, Hermann
Update on drug-drug interaction at organic cation transporters: mechanisms, clinical impact, and proposal for advanced in vitro testing Journal Article
In: vol. 17, no. 6, pp. 635–653, 2021, ISSN: 1742-5255, (Publisher: Taylor & Francis _eprint: https://doi.org/10.1080/17425255.2021.1915284).
Abstract | Links | BibTeX | Tags: {MATE}1, {MATE}2-K, {OCT}1, {OCT}2, {OCT}3, {PMAT}, Drug-drug interaction, high affinity inhibition, in vitro testing, organic cation transport
@article{koepsell_update_2021,
title = {Update on drug-drug interaction at organic cation transporters: mechanisms, clinical impact, and proposal for advanced in vitro testing},
author = {Hermann Koepsell},
url = {https://doi.org/10.1080/17425255.2021.1915284},
doi = {10.1080/17425255.2021.1915284},
issn = {1742-5255},
year = {2021},
date = {2021-06-03},
urldate = {2024-06-04},
volume = {17},
number = {6},
pages = {635–653},
abstract = {Introduction: Organic cation transporters collectively called OCTs belong to three gene families (SLC22A1 OCT1, SLC22A2 OCT2, SLC22A3 OCT3, SLC22A4 OCTN1, SLC22A5 OCTN2, SLC29A4 PMAT, SLC47A1 MATE1, and SLC47A1 MATE2-K). OCTs transport structurally diverse drugs with overlapping selectivity. Some OCTs were shown to be critically involved in pharmacokinetics and therapeutic efficacy of cationic drugs. Drug-drug interactions at individual OCTs were shown to result in clinical effects. Procedures for in vitro testing of drugs for interaction with OCT1, OCT2, MATE1, and MATE2-K have been recommended. Areas covered: An overview of functional properties, cation selectivity, location, and clinical impact of OCTs is provided. In addition, clinically relevant drug-drug interactions in OCTs are compiled. Because it was observed that the half maximal concentration of drugs to inhibit transport by OCTs (IC50) is dependent on the transported cation and its concentration, an advanced protocol for in vitro testing of drugs for interaction with OCTs is proposed. In addition, it is suggested to include OCT3 and PMAT for in vitro testing. Expert opinion: Research on clinical roles of OCTs should be reinforced including more transporters and drugs. An improvement of the in vitro testing protocol considering recent data is imperative for the benefit of patients.},
note = {Publisher: Taylor & Francis
_eprint: https://doi.org/10.1080/17425255.2021.1915284},
keywords = {{MATE}1, {MATE}2-K, {OCT}1, {OCT}2, {OCT}3, {PMAT}, Drug-drug interaction, high affinity inhibition, in vitro testing, organic cation transport},
pubstate = {published},
tppubtype = {article}
}
Introduction: Organic cation transporters collectively called OCTs belong to three gene families (SLC22A1 OCT1, SLC22A2 OCT2, SLC22A3 OCT3, SLC22A4 OCTN1, SLC22A5 OCTN2, SLC29A4 PMAT, SLC47A1 MATE1, and SLC47A1 MATE2-K). OCTs transport structurally diverse drugs with overlapping selectivity. Some OCTs were shown to be critically involved in pharmacokinetics and therapeutic efficacy of cationic drugs. Drug-drug interactions at individual OCTs were shown to result in clinical effects. Procedures for in vitro testing of drugs for interaction with OCT1, OCT2, MATE1, and MATE2-K have been recommended. Areas covered: An overview of functional properties, cation selectivity, location, and clinical impact of OCTs is provided. In addition, clinically relevant drug-drug interactions in OCTs are compiled. Because it was observed that the half maximal concentration of drugs to inhibit transport by OCTs (IC50) is dependent on the transported cation and its concentration, an advanced protocol for in vitro testing of drugs for interaction with OCTs is proposed. In addition, it is suggested to include OCT3 and PMAT for in vitro testing. Expert opinion: Research on clinical roles of OCTs should be reinforced including more transporters and drugs. An improvement of the in vitro testing protocol considering recent data is imperative for the benefit of patients.
Raimondi, Stefano; Musmeci, Eliana; Candeliere, Francesco; Amaretti, Alberto; Rossi, Maddalena
Identification of mucin degraders of the human gut microbiota Journal Article
In: vol. 11, no. 1, pp. 11094, 2021, ISSN: 2045-2322, (Number: 1 Publisher: Nature Publishing Group).
Abstract | Links | BibTeX | Tags: Bacteria, Microbial communities, Microbiology
@article{raimondi_identification_2021,
title = {Identification of mucin degraders of the human gut microbiota},
author = {Stefano Raimondi and Eliana Musmeci and Francesco Candeliere and Alberto Amaretti and Maddalena Rossi},
url = {https://www.nature.com/articles/s41598-021-90553-4},
doi = {10.1038/s41598-021-90553-4},
issn = {2045-2322},
year = {2021},
date = {2021-05-27},
urldate = {2023-02-23},
volume = {11},
number = {1},
pages = {11094},
abstract = {Mucins are large glycoproteins consisting of approximately 80% of hetero-oligosaccharides. Gut mucin degraders of healthy subjects were investigated, through a culture dependent and independent approach. The faeces of five healthy adults were subjected to three steps of anaerobic enrichment in a medium with sole mucins as carbon and nitrogen sources. The bacterial community was compared before and after the enrichment by 16S rRNA gene profiling. Bacteria capable of fermenting sugars, such as Anaerotruncus, Holdemania, and Enterococcaceae likely took advantage of the carbohydrate chains. Escherichia coli and Enterobacteriaceae, Peptococcales, the Coriobacteriale Eggerthella, and a variety of Clostridia such as Oscillospiraceae, Anaerotruncus, and Lachnoclostridium, significantly increased and likely participated to the degradation of the protein backbone of mucin. The affinity of E. coli and Enterobacteriaceae for mucin may facilitate the access to the gut mucosa, promoting gut barrier damage and triggering systemic inflammatory responses. Only three species of strict anaerobes able to grow on mucin were isolated from the enrichments of five different microbiota: Clostridium disporicum, Clostridium tertium, and Paraclostridium benzoelyticum. The limited number of species isolated confirms that in the gut the degradation of these glycoproteins results from cooperation and cross-feeding among several species exhibiting different metabolic capabilities.},
note = {Number: 1
Publisher: Nature Publishing Group},
keywords = {Bacteria, Microbial communities, Microbiology},
pubstate = {published},
tppubtype = {article}
}
Mucins are large glycoproteins consisting of approximately 80% of hetero-oligosaccharides. Gut mucin degraders of healthy subjects were investigated, through a culture dependent and independent approach. The faeces of five healthy adults were subjected to three steps of anaerobic enrichment in a medium with sole mucins as carbon and nitrogen sources. The bacterial community was compared before and after the enrichment by 16S rRNA gene profiling. Bacteria capable of fermenting sugars, such as Anaerotruncus, Holdemania, and Enterococcaceae likely took advantage of the carbohydrate chains. Escherichia coli and Enterobacteriaceae, Peptococcales, the Coriobacteriale Eggerthella, and a variety of Clostridia such as Oscillospiraceae, Anaerotruncus, and Lachnoclostridium, significantly increased and likely participated to the degradation of the protein backbone of mucin. The affinity of E. coli and Enterobacteriaceae for mucin may facilitate the access to the gut mucosa, promoting gut barrier damage and triggering systemic inflammatory responses. Only three species of strict anaerobes able to grow on mucin were isolated from the enrichments of five different microbiota: Clostridium disporicum, Clostridium tertium, and Paraclostridium benzoelyticum. The limited number of species isolated confirms that in the gut the degradation of these glycoproteins results from cooperation and cross-feeding among several species exhibiting different metabolic capabilities.
Song, Yun-Qing; He, Rong-Jing; Pu, Dan; Guan, Xiao-Qing; Shi, Jin-Hui; Li, Yao-Guang; Hou, Jie; Jia, Shou-Ning; Qin, Wei-Wei; Fang, Sheng-Quan; Ge, Guang-Bo
Discovery and Characterization of the Biflavones From Ginkgo biloba as Highly Specific and Potent Inhibitors Against Human Carboxylesterase 2 Journal Article
In: vol. 12, pp. 655659, 2021, ISSN: 1663-9812.
Abstract | Links | BibTeX | Tags:
@article{song_discovery_2021,
title = {Discovery and Characterization of the Biflavones From Ginkgo biloba as Highly Specific and Potent Inhibitors Against Human Carboxylesterase 2},
author = {Yun-Qing Song and Rong-Jing He and Dan Pu and Xiao-Qing Guan and Jin-Hui Shi and Yao-Guang Li and Jie Hou and Shou-Ning Jia and Wei-Wei Qin and Sheng-Quan Fang and Guang-Bo Ge},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8167799/},
doi = {10.3389/fphar.2021.655659},
issn = {1663-9812},
year = {2021},
date = {2021-05-18},
urldate = {2025-05-02},
volume = {12},
pages = {655659},
abstract = {Human carboxylesterase 2 (CES2), one of the most abundant hydrolases distributed in the small intestine, has been validated as a key therapeutic target to ameliorate the intestinal toxicity caused by irinotecan. This study aims to discover efficacious CES2 inhibitors from natural products and to characterize the inhibition potentials and inhibitory mechanisms of the newly identified CES2 inhibitors. Following high-throughput screening and evaluation of the inhibition potency of more than 100 natural products against CES2, it was found that the biflavones isolated from Ginkgo biloba displayed extremely potent CES2 inhibition activities and high specificity over CES1 (>1000-fold). Further investigation showed that ginkgetin, bilobetin, sciadopitysin and isoginkgetin potently inhibited CES2-catalyzed hydrolysis of various substrates, including the CES2 substrate-drug irinotecan. Notably, the inhibition potentials of four biflavones against CES2 were more potent than that of loperamide, a marketed anti-diarrhea agent used for alleviating irinotecan-induced intestinal toxicity. Inhibition kinetic analyses demonstrated that ginkgetin, bilobetin, sciadopitysin and isoginkgetin potently inhibited CES2-catalyzed fluorescein diacetate hydrolysis via a reversible and mixed inhibition manner, with K
i values of less than 100 nM. Ensemble docking and molecular dynamics revealed that these biflavones could tightly and stably bind on the catalytic cavity of CES2 via hydrogen bonding and π-π stacking interactions, while the interactions with CES1 were awfully poor. Collectively, this study reports that the biflavones isolated from Ginkgo biloba are potent and highly specific CES2 inhibitors, which offers several promising lead compounds for developing novel anti-diarrhea agent to alleviate irinotecan-induced diarrhea.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Human carboxylesterase 2 (CES2), one of the most abundant hydrolases distributed in the small intestine, has been validated as a key therapeutic target to ameliorate the intestinal toxicity caused by irinotecan. This study aims to discover efficacious CES2 inhibitors from natural products and to characterize the inhibition potentials and inhibitory mechanisms of the newly identified CES2 inhibitors. Following high-throughput screening and evaluation of the inhibition potency of more than 100 natural products against CES2, it was found that the biflavones isolated from Ginkgo biloba displayed extremely potent CES2 inhibition activities and high specificity over CES1 (>1000-fold). Further investigation showed that ginkgetin, bilobetin, sciadopitysin and isoginkgetin potently inhibited CES2-catalyzed hydrolysis of various substrates, including the CES2 substrate-drug irinotecan. Notably, the inhibition potentials of four biflavones against CES2 were more potent than that of loperamide, a marketed anti-diarrhea agent used for alleviating irinotecan-induced intestinal toxicity. Inhibition kinetic analyses demonstrated that ginkgetin, bilobetin, sciadopitysin and isoginkgetin potently inhibited CES2-catalyzed fluorescein diacetate hydrolysis via a reversible and mixed inhibition manner, with K
i values of less than 100 nM. Ensemble docking and molecular dynamics revealed that these biflavones could tightly and stably bind on the catalytic cavity of CES2 via hydrogen bonding and π-π stacking interactions, while the interactions with CES1 were awfully poor. Collectively, this study reports that the biflavones isolated from Ginkgo biloba are potent and highly specific CES2 inhibitors, which offers several promising lead compounds for developing novel anti-diarrhea agent to alleviate irinotecan-induced diarrhea.
i values of less than 100 nM. Ensemble docking and molecular dynamics revealed that these biflavones could tightly and stably bind on the catalytic cavity of CES2 via hydrogen bonding and π-π stacking interactions, while the interactions with CES1 were awfully poor. Collectively, this study reports that the biflavones isolated from Ginkgo biloba are potent and highly specific CES2 inhibitors, which offers several promising lead compounds for developing novel anti-diarrhea agent to alleviate irinotecan-induced diarrhea.
Dutton, Johanna S; Hinman, Samuel S; Kim, Raehyun; Attayek, Peter J; Maurer, Mallory; Sims, Christopher S; Allbritton, Nancy L
Hyperglycemia minimally alters primary self-renewing human colonic epithelial cells while TNFα-promotes severe intestinal epithelial dysfunction Journal Article
In: vol. 13, no. 6, pp. 139–152, 2021, ISSN: 1757-9694.
Abstract | Links | BibTeX | Tags:
@article{dutton_hyperglycemia_2021,
title = {Hyperglycemia minimally alters primary self-renewing human colonic epithelial cells while TNFα-promotes severe intestinal epithelial dysfunction},
author = {Johanna S Dutton and Samuel S Hinman and Raehyun Kim and Peter J Attayek and Mallory Maurer and Christopher S Sims and Nancy L Allbritton},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8204630/},
doi = {10.1093/intbio/zyab008},
issn = {1757-9694},
year = {2021},
date = {2021-05-15},
urldate = {2023-07-03},
volume = {13},
number = {6},
pages = {139–152},
abstract = {Hyperglycemia is thought to increase production of inflammatory cytokines and permeability of the large intestine. Resulting intestinal inflammation is then often characterized by excess secretion of tumor necrosis factor alpha (TNFα). Thus, hyperglycemia in hospitalized patients suffering from severe trauma or disease is frequently accompanied by TNFα secretion, and the combined impact of these insults on the intestinal epithelium is poorly understood. This study utilized a simple yet elegant model of the intestinal epithelium, comprised of primary human intestinal stem cells and their differentiated progeny, to investigate the impact of hyperglycemia and inflammatory factors on the colonic epithelium. When compared to epithelium cultured under conditions of physiologic glucose, cells under hyperglycemic conditions displayed decreased mucin-2 (MUC2), as well as diminished alkaline phosphatase (ALP) activity. Conditions of 60 mM glucose potentiated secretion of the cytokine IL-8 suggesting that cytokine secretion during hyperglycemia may be a source of tissue inflammation. TNFα measurably increased secretion of IL-8 and IL-1β, which was enhanced at 60 mM glucose. Surprisingly, intestinal permeability and paracellular transport were not altered by even extreme levels of hyperglycemia. The presence of TNFα increased MUC2 presence, decreased ALP activity, and negatively impacted monolayer barrier function. When TNFα hyperglycemia and ≤30 mM glucose and were combined, MUC2 and ALP activity remained similar to that of TNFα alone, although synergistic effects were seen at 60 mM glucose. An automated image analysis pipeline was developed to assay changes in properties of the zonula occludens-1 (ZO-1)-demarcated cell boundaries. While hyperglycemia alone had little impact on cell shape and size, cell morphologic properties were extraordinarily sensitive to soluble TNFα. These results suggest that TNFα acted as the dominant modulator of the epithelium relative to glucose, and that control of inflammation rather than glucose may be key to maintaining intestinal homeostasis.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Hyperglycemia is thought to increase production of inflammatory cytokines and permeability of the large intestine. Resulting intestinal inflammation is then often characterized by excess secretion of tumor necrosis factor alpha (TNFα). Thus, hyperglycemia in hospitalized patients suffering from severe trauma or disease is frequently accompanied by TNFα secretion, and the combined impact of these insults on the intestinal epithelium is poorly understood. This study utilized a simple yet elegant model of the intestinal epithelium, comprised of primary human intestinal stem cells and their differentiated progeny, to investigate the impact of hyperglycemia and inflammatory factors on the colonic epithelium. When compared to epithelium cultured under conditions of physiologic glucose, cells under hyperglycemic conditions displayed decreased mucin-2 (MUC2), as well as diminished alkaline phosphatase (ALP) activity. Conditions of 60 mM glucose potentiated secretion of the cytokine IL-8 suggesting that cytokine secretion during hyperglycemia may be a source of tissue inflammation. TNFα measurably increased secretion of IL-8 and IL-1β, which was enhanced at 60 mM glucose. Surprisingly, intestinal permeability and paracellular transport were not altered by even extreme levels of hyperglycemia. The presence of TNFα increased MUC2 presence, decreased ALP activity, and negatively impacted monolayer barrier function. When TNFα hyperglycemia and ≤30 mM glucose and were combined, MUC2 and ALP activity remained similar to that of TNFα alone, although synergistic effects were seen at 60 mM glucose. An automated image analysis pipeline was developed to assay changes in properties of the zonula occludens-1 (ZO-1)-demarcated cell boundaries. While hyperglycemia alone had little impact on cell shape and size, cell morphologic properties were extraordinarily sensitive to soluble TNFα. These results suggest that TNFα acted as the dominant modulator of the epithelium relative to glucose, and that control of inflammation rather than glucose may be key to maintaining intestinal homeostasis.
Sodhi, Jasleen K.; Benet, Leslie Z.
Successful and Unsuccessful Prediction of Human Hepatic Clearance for Lead Optimization Journal Article
In: vol. 64, no. 7, pp. 3546–3559, 2021, ISSN: 0022-2623.
Abstract | Links | BibTeX | Tags:
@article{sodhi_successful_2021,
title = {Successful and Unsuccessful Prediction of Human Hepatic Clearance for Lead Optimization},
author = {Jasleen K. Sodhi and Leslie Z. Benet},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8504179/},
doi = {10.1021/acs.jmedchem.0c01930},
issn = {0022-2623},
year = {2021},
date = {2021-04-08},
urldate = {2025-05-12},
volume = {64},
number = {7},
pages = {3546–3559},
abstract = {Development of new chemical entities is costly, time-consuming, and has a low success rate. Accurate prediction of pharmacokinetic properties is critical to progress compounds with favorable drug-like characteristics in lead optimization. Of particular importance is the prediction of hepatic clearance, which determines drug exposure and contributes to projection of dose, half-life, and bioavailability. The most commonly employed methodology to predict hepatic clearance is termed in vitro to in vivo extrapolation (IVIVE) that involves measuring drug metabolism in vitro, scaling-up this in vitro intrinsic clearance to a prediction of in vivo intrinsic clearance by reconciling the enzymatic content between the incubation and an average human liver, and applying a model of hepatic disposition to account for limitations of protein binding and blood flow to predict in vivo clearance. This manuscript reviews common in vitro techniques used to predict hepatic clearance as well as current challenges and recent theoretical advancements in IVIVE.,},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Development of new chemical entities is costly, time-consuming, and has a low success rate. Accurate prediction of pharmacokinetic properties is critical to progress compounds with favorable drug-like characteristics in lead optimization. Of particular importance is the prediction of hepatic clearance, which determines drug exposure and contributes to projection of dose, half-life, and bioavailability. The most commonly employed methodology to predict hepatic clearance is termed in vitro to in vivo extrapolation (IVIVE) that involves measuring drug metabolism in vitro, scaling-up this in vitro intrinsic clearance to a prediction of in vivo intrinsic clearance by reconciling the enzymatic content between the incubation and an average human liver, and applying a model of hepatic disposition to account for limitations of protein binding and blood flow to predict in vivo clearance. This manuscript reviews common in vitro techniques used to predict hepatic clearance as well as current challenges and recent theoretical advancements in IVIVE.,
Janssen, Aafke W. F.; Duivenvoorde, Loes P. M.; Rijkers, Deborah; Nijssen, Rosalie; Peijnenburg, Ad A. C. M.; Zande, Meike; Louisse, Jochem
In: vol. 95, no. 3, pp. 907–922, 2021, ISSN: 1432-0738.
Abstract | Links | BibTeX | Tags: Cytochrome P450 ({CYP}), Gastrointestinal tract, Organotypic models, Stem Cells, Toxicology
@article{janssen_cytochrome_2021,
title = {Cytochrome P450 expression, induction and activity in human induced pluripotent stem cell-derived intestinal organoids and comparison with primary human intestinal epithelial cells and Caco-2 cells},
author = {Aafke W. F. Janssen and Loes P. M. Duivenvoorde and Deborah Rijkers and Rosalie Nijssen and Ad A. C. M. Peijnenburg and Meike Zande and Jochem Louisse},
url = {https://doi.org/10.1007/s00204-020-02953-6},
doi = {10.1007/s00204-020-02953-6},
issn = {1432-0738},
year = {2021},
date = {2021-03-01},
urldate = {2022-12-01},
volume = {95},
number = {3},
pages = {907–922},
abstract = {Human intestinal organoids (HIOs) are a promising in vitro model consisting of different intestinal cell types with a 3D microarchitecture resembling native tissue. In the current study, we aimed to assess the expression of the most common intestinal CYP enzymes in a human induced pluripotent stem cell (hiPSC)-derived HIO model, and the suitability of that model to study chemical-induced changes in CYP expression and activity. We compared this model with the commonly used human colonic adenocarcinoma cell line Caco-2 and with a human primary intestinal epithelial cell (IEC)-based model, closely resembling in vivo tissue. We optimized an existing protocol to differentiate hiPSCs into HIOs and demonstrated that obtained HIOs contain a polarized epithelium with tight junctions consisting of enterocytes, goblet cells, enteroendocrine cells and Paneth cells. We extensively characterized the gene expression of CYPs and activity of CYP3A4/5, indicating relatively high gene expression levels of the most important intestinal CYP enzymes in HIOs compared to the other models. Furthermore, we showed that CYP1A1 and CYP1B1 were induced by β-naphtoflavone in all three models, whereas CYP3A4 was induced by phenobarbital and rifampicin in HIOs, in the IEC-based model (although not statistically significant), but not in Caco-2 cells. Interestingly, CYP2B6 expression was not induced in any of the models by the well-known liver CYP2B6 inducer phenobarbital. In conclusion, our study indicates that hiPSC-based HIOs are a useful in vitro intestinal model to study biotransformation of chemicals in the intestine.},
keywords = {Cytochrome P450 ({CYP}), Gastrointestinal tract, Organotypic models, Stem Cells, Toxicology},
pubstate = {published},
tppubtype = {article}
}
Human intestinal organoids (HIOs) are a promising in vitro model consisting of different intestinal cell types with a 3D microarchitecture resembling native tissue. In the current study, we aimed to assess the expression of the most common intestinal CYP enzymes in a human induced pluripotent stem cell (hiPSC)-derived HIO model, and the suitability of that model to study chemical-induced changes in CYP expression and activity. We compared this model with the commonly used human colonic adenocarcinoma cell line Caco-2 and with a human primary intestinal epithelial cell (IEC)-based model, closely resembling in vivo tissue. We optimized an existing protocol to differentiate hiPSCs into HIOs and demonstrated that obtained HIOs contain a polarized epithelium with tight junctions consisting of enterocytes, goblet cells, enteroendocrine cells and Paneth cells. We extensively characterized the gene expression of CYPs and activity of CYP3A4/5, indicating relatively high gene expression levels of the most important intestinal CYP enzymes in HIOs compared to the other models. Furthermore, we showed that CYP1A1 and CYP1B1 were induced by β-naphtoflavone in all three models, whereas CYP3A4 was induced by phenobarbital and rifampicin in HIOs, in the IEC-based model (although not statistically significant), but not in Caco-2 cells. Interestingly, CYP2B6 expression was not induced in any of the models by the well-known liver CYP2B6 inducer phenobarbital. In conclusion, our study indicates that hiPSC-based HIOs are a useful in vitro intestinal model to study biotransformation of chemicals in the intestine.
Markus, Jan; Landry, Tim; Stevens, Zachary; Scott, Hailey; Llanos, Pierre; Debatis, Michelle; Armento, Alexander; Klausner, Mitchell; Ayehunie, Seyoum
Human small intestinal organotypic culture model for drug permeation, inflammation, and toxicity assays Journal Article
In: vol. 57, no. 2, pp. 160–173, 2021, ISSN: 1543-706X.
Abstract | Links | BibTeX | Tags:
@article{markus_human_2021,
title = {Human small intestinal organotypic culture model for drug permeation, inflammation, and toxicity assays},
author = {Jan Markus and Tim Landry and Zachary Stevens and Hailey Scott and Pierre Llanos and Michelle Debatis and Alexander Armento and Mitchell Klausner and Seyoum Ayehunie},
url = {https://doi.org/10.1007/s11626-020-00526-6},
doi = {10.1007/s11626-020-00526-6},
issn = {1543-706X},
year = {2021},
date = {2021-02-01},
urldate = {2023-02-20},
volume = {57},
number = {2},
pages = {160–173},
abstract = {The gastrointestinal tract (GIT), in particular, the small intestine, plays a significant role in food digestion, fluid and electrolyte transport, drug absorption and metabolism, and nutrient uptake. As the longest portion of the GIT, the small intestine also plays a vital role in protecting the host against pathogenic or opportunistic microbial invasion. However, establishing polarized intestinal tissue models in vitro that reflect the architecture and physiology of the gut has been a challenge for decades and the lack of translational models that predict human responses has impeded research in the drug absorption, metabolism, and drug-induced gastrointestinal toxicity space. Often, animals fail to recapitulate human physiology and do not predict human outcomes. Also, certain human pathogens are species specific and do not infect other hosts. Concerns such as variability of results, a low throughput format, and ethical considerations further complicate the use of animals for predicting the safety and efficacy xenobiotics in humans. These limitations necessitate the development of in vitro 3D human intestinal tissue models that recapitulate in vivo–like microenvironment and provide more physiologically relevant cellular responses so that they can better predict the safety and efficacy of pharmaceuticals and toxicants. Over the past decade, much progress has been made in the development of in vitro intestinal models (organoids and 3D-organotypic tissues) using either inducible pluripotent or adult stem cells. Among the models, the MatTek’s intestinal tissue model (EpiIntestinal™ Ashland, MA) has been used extensively by the pharmaceutical industry to study drug permeation, metabolism, drug-induced GI toxicity, pathogen infections, inflammation, wound healing, and as a predictive model for a clinical adverse outcome (diarrhea) to pharmaceutical drugs. In this paper, our review will focus on the potential of in vitro small intestinal tissues as preclinical research tool and as alternative to the use of animals.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The gastrointestinal tract (GIT), in particular, the small intestine, plays a significant role in food digestion, fluid and electrolyte transport, drug absorption and metabolism, and nutrient uptake. As the longest portion of the GIT, the small intestine also plays a vital role in protecting the host against pathogenic or opportunistic microbial invasion. However, establishing polarized intestinal tissue models in vitro that reflect the architecture and physiology of the gut has been a challenge for decades and the lack of translational models that predict human responses has impeded research in the drug absorption, metabolism, and drug-induced gastrointestinal toxicity space. Often, animals fail to recapitulate human physiology and do not predict human outcomes. Also, certain human pathogens are species specific and do not infect other hosts. Concerns such as variability of results, a low throughput format, and ethical considerations further complicate the use of animals for predicting the safety and efficacy xenobiotics in humans. These limitations necessitate the development of in vitro 3D human intestinal tissue models that recapitulate in vivo–like microenvironment and provide more physiologically relevant cellular responses so that they can better predict the safety and efficacy of pharmaceuticals and toxicants. Over the past decade, much progress has been made in the development of in vitro intestinal models (organoids and 3D-organotypic tissues) using either inducible pluripotent or adult stem cells. Among the models, the MatTek’s intestinal tissue model (EpiIntestinal™ Ashland, MA) has been used extensively by the pharmaceutical industry to study drug permeation, metabolism, drug-induced GI toxicity, pathogen infections, inflammation, wound healing, and as a predictive model for a clinical adverse outcome (diarrhea) to pharmaceutical drugs. In this paper, our review will focus on the potential of in vitro small intestinal tissues as preclinical research tool and as alternative to the use of animals.
Apostolou, Athanasia; Panchakshari, Rohit A.; Banerjee, Antara; Manatakis, Dimitris V.; Paraskevopoulou, Maria D.; Luc, Raymond; Abu-Ali, Galeb; Dimitriou, Alexandra; Lucchesi, Carolina; Kulkarni, Gauri; Maulana, Tengku Ibrahim; Kasendra, Magdalena; Kerns, Jordan S.; Bleck, Bertram; Ewart, Lorna; Manolakos, Elias S.; Hamilton, Geraldine A.; Giallourakis, Cosmas; Karalis, Katia
A Novel Microphysiological Colon Platform to Decipher Mechanisms Driving Human Intestinal Permeability Journal Article
In: vol. 12, no. 5, pp. 1719–1741, 2021, ISSN: 2352-345X.
Abstract | Links | BibTeX | Tags: Interleukin 22, Leaky Gut, Organ-on-Chip, Organoids
@article{apostolou_novel_2021,
title = {A Novel Microphysiological Colon Platform to Decipher Mechanisms Driving Human Intestinal Permeability},
author = {Athanasia Apostolou and Rohit A. Panchakshari and Antara Banerjee and Dimitris V. Manatakis and Maria D. Paraskevopoulou and Raymond Luc and Galeb Abu-Ali and Alexandra Dimitriou and Carolina Lucchesi and Gauri Kulkarni and Tengku Ibrahim Maulana and Magdalena Kasendra and Jordan S. Kerns and Bertram Bleck and Lorna Ewart and Elias S. Manolakos and Geraldine A. Hamilton and Cosmas Giallourakis and Katia Karalis},
url = {https://www.sciencedirect.com/science/article/pii/S2352345X21001454},
doi = {10.1016/j.jcmgh.2021.07.004},
issn = {2352-345X},
year = {2021},
date = {2021-01-01},
urldate = {2024-07-31},
volume = {12},
number = {5},
pages = {1719–1741},
abstract = {Background & Aims
The limited availability of organoid systems that mimic the molecular signatures and architecture of human intestinal epithelium has been an impediment to allowing them to be harnessed for the development of therapeutics as well as physiological insights. We developed a microphysiological Organ-on-Chip (Emulate, Inc, Boston, MA) platform designed to mimic properties of human intestinal epithelium leading to insights into barrier integrity.
Methods
We combined the human biopsy-derived leucine-rich repeat-containing G-protein–coupled receptor 5–positive organoids and Organ-on-Chip technologies to establish a micro-engineered human Colon Intestine-Chip (Emulate, Inc, Boston, MA). We characterized the proximity of the model to human tissue and organoids maintained in suspension by RNA sequencing analysis, and their differentiation to intestinal epithelial cells on the Colon Intestine-Chip under variable conditions. Furthermore, organoids from different donors were evaluated to understand variability in the system. Our system was applied to understanding the epithelial barrier and characterizing mechanisms driving the cytokine-induced barrier disruption.
Results
Our data highlight the importance of the endothelium and the in vivo tissue-relevant dynamic microenvironment in the Colon Intestine-Chip in the establishment of a tight monolayer of differentiated, polarized, organoid-derived intestinal epithelial cells. We confirmed the effect of interferon-γ on the colonic barrier and identified reorganization of apical junctional complexes, and induction of apoptosis in the intestinal epithelial cells as mediating mechanisms. We show that in the human Colon Intestine-Chip exposure to interleukin 22 induces disruption of the barrier, unlike its described protective role in experimental colitis in mice.
Conclusions
We developed a human Colon Intestine-Chip platform and showed its value in the characterization of the mechanism of action of interleukin 22 in the human epithelial barrier. This system can be used to elucidate, in a time- and challenge-dependent manner, the mechanism driving the development of leaky gut in human beings and to identify associated biomarkers.},
keywords = {Interleukin 22, Leaky Gut, Organ-on-Chip, Organoids},
pubstate = {published},
tppubtype = {article}
}
Background & Aims
The limited availability of organoid systems that mimic the molecular signatures and architecture of human intestinal epithelium has been an impediment to allowing them to be harnessed for the development of therapeutics as well as physiological insights. We developed a microphysiological Organ-on-Chip (Emulate, Inc, Boston, MA) platform designed to mimic properties of human intestinal epithelium leading to insights into barrier integrity.
Methods
We combined the human biopsy-derived leucine-rich repeat-containing G-protein–coupled receptor 5–positive organoids and Organ-on-Chip technologies to establish a micro-engineered human Colon Intestine-Chip (Emulate, Inc, Boston, MA). We characterized the proximity of the model to human tissue and organoids maintained in suspension by RNA sequencing analysis, and their differentiation to intestinal epithelial cells on the Colon Intestine-Chip under variable conditions. Furthermore, organoids from different donors were evaluated to understand variability in the system. Our system was applied to understanding the epithelial barrier and characterizing mechanisms driving the cytokine-induced barrier disruption.
Results
Our data highlight the importance of the endothelium and the in vivo tissue-relevant dynamic microenvironment in the Colon Intestine-Chip in the establishment of a tight monolayer of differentiated, polarized, organoid-derived intestinal epithelial cells. We confirmed the effect of interferon-γ on the colonic barrier and identified reorganization of apical junctional complexes, and induction of apoptosis in the intestinal epithelial cells as mediating mechanisms. We show that in the human Colon Intestine-Chip exposure to interleukin 22 induces disruption of the barrier, unlike its described protective role in experimental colitis in mice.
Conclusions
We developed a human Colon Intestine-Chip platform and showed its value in the characterization of the mechanism of action of interleukin 22 in the human epithelial barrier. This system can be used to elucidate, in a time- and challenge-dependent manner, the mechanism driving the development of leaky gut in human beings and to identify associated biomarkers.
The limited availability of organoid systems that mimic the molecular signatures and architecture of human intestinal epithelium has been an impediment to allowing them to be harnessed for the development of therapeutics as well as physiological insights. We developed a microphysiological Organ-on-Chip (Emulate, Inc, Boston, MA) platform designed to mimic properties of human intestinal epithelium leading to insights into barrier integrity.
Methods
We combined the human biopsy-derived leucine-rich repeat-containing G-protein–coupled receptor 5–positive organoids and Organ-on-Chip technologies to establish a micro-engineered human Colon Intestine-Chip (Emulate, Inc, Boston, MA). We characterized the proximity of the model to human tissue and organoids maintained in suspension by RNA sequencing analysis, and their differentiation to intestinal epithelial cells on the Colon Intestine-Chip under variable conditions. Furthermore, organoids from different donors were evaluated to understand variability in the system. Our system was applied to understanding the epithelial barrier and characterizing mechanisms driving the cytokine-induced barrier disruption.
Results
Our data highlight the importance of the endothelium and the in vivo tissue-relevant dynamic microenvironment in the Colon Intestine-Chip in the establishment of a tight monolayer of differentiated, polarized, organoid-derived intestinal epithelial cells. We confirmed the effect of interferon-γ on the colonic barrier and identified reorganization of apical junctional complexes, and induction of apoptosis in the intestinal epithelial cells as mediating mechanisms. We show that in the human Colon Intestine-Chip exposure to interleukin 22 induces disruption of the barrier, unlike its described protective role in experimental colitis in mice.
Conclusions
We developed a human Colon Intestine-Chip platform and showed its value in the characterization of the mechanism of action of interleukin 22 in the human epithelial barrier. This system can be used to elucidate, in a time- and challenge-dependent manner, the mechanism driving the development of leaky gut in human beings and to identify associated biomarkers.
2020
Li, Albert P.
In: vol. 48, no. 10, pp. 980–992, 2020, ISSN: 0090-9556, 1521-009X, (Publisher: American Society for Pharmacology and Experimental Therapeutics Section: Special Section on Natural Products: Experimental Approaches to Elucidate Disposition Mechanisms and Predict Pharmacokinetic Drug Interactions — Minireview).
Abstract | Links | BibTeX | Tags:
@article{li_vitro_2020,
title = {In Vitro Human Cell–Based Experimental Models for the Evaluation of Enteric Metabolism and Drug Interaction Potential of Drugs and Natural Products},
author = {Albert P. Li},
url = {https://dmd.aspetjournals.org/content/48/10/980},
doi = {10.1124/dmd.120.000053},
issn = {0090-9556, 1521-009X},
year = {2020},
date = {2020-10-01},
urldate = {2023-06-16},
volume = {48},
number = {10},
pages = {980–992},
abstract = {Elements of key enteric drug metabolism and disposition pathways are reviewed to aid the assessment of the applicability of current cell-based enteric experimental systems for the evaluation of enteric metabolism and drug interaction potential. Enteric nuclear receptors include vitamin D receptor, constitutive androstane receptor, pregnane X receptor, farnesoid X receptor, liver X receptor, aryl hydrocarbon receptor, and peroxisome proliferator–activated receptor. Enteric drug metabolizing enzyme pathways include both cytochrome P450 (P450) and non-P450 drug metabolizing enzymes based on gene expression, proteomics, and activity. Both uptake and efflux transporters are present in the small intestine, with P-glycoprotein found to be responsible for most drug-drug and food-drug interactions. The cell-based in vitro enteric systems reviewed are 1) immortalized cell line model: the human colon adenocarcinoma (Caco-2) cells; 2) human stem cell–derived enterocyte models: stem cell enteric systems, either from intestinal crypt cells or induced pluripotent stem cells; and 3) primary cell models: human intestinal slices, cryopreserved human enterocytes, permeabilized cofactor-supplemented (MetMax) cryopreserved human enterocytes, and cryopreserved human intestinal mucosa. The major deficiency with both immortalized cell lines and stem cell–derived enterocytes is that drug metabolizing enzyme activities, although they are detectable, are substantially lower than those for the intestinal mucosa in vivo. Human intestine slices, cryopreserved human enterocytes, MetMax cryopreserved human enterocytes, and cryopreserved human intestinal mucosa retain robust enteric drug metabolizing enzyme activity and represent appropriate models for the evaluation of metabolism and metabolism-dependent drug interaction potential of orally administered xenobiotics including drugs, botanical products, and dietary supplements.
Significance Statement Enteric drug metabolism plays an important role in the bioavailability and metabolic fate of orally administered drugs as well as in enteric drug-drug and food-drug interactions. The current status of key enteric drug metabolism and disposition pathways and in vitro human cell–based enteric experimental systems for the evaluation of the metabolism and drug interaction potential of orally administered substances is reviewed.},
note = {Publisher: American Society for Pharmacology and Experimental Therapeutics
Section: Special Section on Natural Products: Experimental Approaches to Elucidate Disposition Mechanisms and Predict Pharmacokinetic Drug Interactions — Minireview},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Elements of key enteric drug metabolism and disposition pathways are reviewed to aid the assessment of the applicability of current cell-based enteric experimental systems for the evaluation of enteric metabolism and drug interaction potential. Enteric nuclear receptors include vitamin D receptor, constitutive androstane receptor, pregnane X receptor, farnesoid X receptor, liver X receptor, aryl hydrocarbon receptor, and peroxisome proliferator–activated receptor. Enteric drug metabolizing enzyme pathways include both cytochrome P450 (P450) and non-P450 drug metabolizing enzymes based on gene expression, proteomics, and activity. Both uptake and efflux transporters are present in the small intestine, with P-glycoprotein found to be responsible for most drug-drug and food-drug interactions. The cell-based in vitro enteric systems reviewed are 1) immortalized cell line model: the human colon adenocarcinoma (Caco-2) cells; 2) human stem cell–derived enterocyte models: stem cell enteric systems, either from intestinal crypt cells or induced pluripotent stem cells; and 3) primary cell models: human intestinal slices, cryopreserved human enterocytes, permeabilized cofactor-supplemented (MetMax) cryopreserved human enterocytes, and cryopreserved human intestinal mucosa. The major deficiency with both immortalized cell lines and stem cell–derived enterocytes is that drug metabolizing enzyme activities, although they are detectable, are substantially lower than those for the intestinal mucosa in vivo. Human intestine slices, cryopreserved human enterocytes, MetMax cryopreserved human enterocytes, and cryopreserved human intestinal mucosa retain robust enteric drug metabolizing enzyme activity and represent appropriate models for the evaluation of metabolism and metabolism-dependent drug interaction potential of orally administered xenobiotics including drugs, botanical products, and dietary supplements.
Significance Statement Enteric drug metabolism plays an important role in the bioavailability and metabolic fate of orally administered drugs as well as in enteric drug-drug and food-drug interactions. The current status of key enteric drug metabolism and disposition pathways and in vitro human cell–based enteric experimental systems for the evaluation of the metabolism and drug interaction potential of orally administered substances is reviewed.
Significance Statement Enteric drug metabolism plays an important role in the bioavailability and metabolic fate of orally administered drugs as well as in enteric drug-drug and food-drug interactions. The current status of key enteric drug metabolism and disposition pathways and in vitro human cell–based enteric experimental systems for the evaluation of the metabolism and drug interaction potential of orally administered substances is reviewed.
Belair, David G.; Visconti, Richard J.; Hong, Miyoun; Marella, Mathieu; Peters, Matthew F.; Scott, Clay W.; Kolaja, Kyle L.
Human ileal organoid model recapitulates clinical incidence of diarrhea associated with small molecule drugs Journal Article
In: vol. 68, pp. 104928, 2020, ISSN: 0887-2333.
Abstract | Links | BibTeX | Tags:
@article{belair_human_2020,
title = {Human ileal organoid model recapitulates clinical incidence of diarrhea associated with small molecule drugs},
author = {David G. Belair and Richard J. Visconti and Miyoun Hong and Mathieu Marella and Matthew F. Peters and Clay W. Scott and Kyle L. Kolaja},
url = {https://www.sciencedirect.com/science/article/pii/S0887233320304781},
doi = {10.1016/j.tiv.2020.104928},
issn = {0887-2333},
year = {2020},
date = {2020-10-01},
urldate = {2024-07-31},
volume = {68},
pages = {104928},
abstract = {Drug-induced gastrointestinal toxicity (GIT) is a common treatment-emergent adverse event that can negatively impact dosing, thereby limiting efficacy and treatment options for patients. An in vitro assay of GIT is needed to address patient variability, mimic the microphysiology of the gut, and accurately predict drug-induced GIT. Primary human ileal organoids (termed ‘enteroids’) have proven useful for stimulating intestinal stem cell proliferation and differentiation to multiple cell types present in the gut epithelium. Enteroids have enabled characterization of gut biology and the signaling involved in the pathogenesis of disease. Here, enteroids were differentiated from four healthy human donors and assessed for culture duration-dependent differentiation status by immunostaining for gut epithelial markers lysozyme, chromogranin A, mucin, and sucrase isomaltase. Differentiated enteroids were evaluated with a reference set of 31 drugs exhibiting varying degrees of clinical incidence of diarrhea, a common manifestation of GIT that can be caused by drug-induced thinning of the gut epithelium. An assay examining enteroid viability in response to drug treatment demonstrated 90% accuracy for recapitulating the incidence of drug-induced diarrhea. The human enteroid viability assay developed here presents a promising in vitro model for evaluating drug-induced diarrhea.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Drug-induced gastrointestinal toxicity (GIT) is a common treatment-emergent adverse event that can negatively impact dosing, thereby limiting efficacy and treatment options for patients. An in vitro assay of GIT is needed to address patient variability, mimic the microphysiology of the gut, and accurately predict drug-induced GIT. Primary human ileal organoids (termed ‘enteroids’) have proven useful for stimulating intestinal stem cell proliferation and differentiation to multiple cell types present in the gut epithelium. Enteroids have enabled characterization of gut biology and the signaling involved in the pathogenesis of disease. Here, enteroids were differentiated from four healthy human donors and assessed for culture duration-dependent differentiation status by immunostaining for gut epithelial markers lysozyme, chromogranin A, mucin, and sucrase isomaltase. Differentiated enteroids were evaluated with a reference set of 31 drugs exhibiting varying degrees of clinical incidence of diarrhea, a common manifestation of GIT that can be caused by drug-induced thinning of the gut epithelium. An assay examining enteroid viability in response to drug treatment demonstrated 90% accuracy for recapitulating the incidence of drug-induced diarrhea. The human enteroid viability assay developed here presents a promising in vitro model for evaluating drug-induced diarrhea.
Wang, Yuli; Sims, Christopher E.; Allbritton, Nancy L.
Enterochromaffin Cell-Enriched Monolayer Platform for Assaying Serotonin Release from Human Primary Intestinal Cells Journal Article
In: vol. 92, no. 18, pp. 12330–12337, 2020, ISSN: 1520-6882.
Abstract | Links | BibTeX | Tags: Enterochromaffin Cells, Humans, Intestinal Mucosa, Serotonin
@article{wang_enterochromaffin_2020,
title = {Enterochromaffin Cell-Enriched Monolayer Platform for Assaying Serotonin Release from Human Primary Intestinal Cells},
author = {Yuli Wang and Christopher E. Sims and Nancy L. Allbritton},
doi = {10.1021/acs.analchem.0c02016},
issn = {1520-6882},
year = {2020},
date = {2020-09-15},
volume = {92},
number = {18},
pages = {12330–12337},
abstract = {Enteroendocrine (EE) cells within the intestinal epithelium produce a range of hormones that have key roles in modulating satiety and feeding behavior in humans. The regulation of hormone release from EE cells as a potential therapeutic strategy to treat metabolic disorders is highly sought after by the pharmaceutical industry. However, functional studies are limited by the scarcity of EE cells (or surrogates) in both in vivo and in vitro systems. Enterochromaffin (EC) cells are a subtype of EE cells that produce serotonin (5HT). Here, we explored simple strategies to enrich EC cells in in vitro monolayer systems derived from human primary intestinal stem cells. During differentiation of the monolayers, the EC cell lineage was significantly altered by both the culture method [air-liquid interface (ALI) vs submerged] and the presence of vasoactive intestinal peptide (VIP). Compared with traditional submerged cultures without VIP, VIP-assisted ALI culture significantly boosted the number of EC cells and their 5HT secretion by up to 430 and 390%, respectively. The method also increased the numbers of other subtypes of EE cells such as L cells. Additionally, this method generated monolayers with enhanced barrier integrity, so that directional (basal or apical) 5HT secretion was measurable. For all donor tissues, the enriched EC cells improved the signal-to-background ratio and reliability of 5HT release assays. The enhancement in the 5HT secretion behavior was consistent over time from a single donor, but significant variation in the amount of secreted 5HT was present among tissues derived from five different donors. To demonstrate the utility of the EC-enriched monolayer system, 13 types of pungent food ingredients were screened for their ability to stimulate 5HT secretion. Curcumin found in the spice turmeric derived from the Curcuma longa plant was found to be the most potent secretagogue. This EC-enriched cell monolayer platform can provide a valuable analytical tool for the high-throughput screening of nutrients and gut microbial components that alter the secretion of 5HT.},
keywords = {Enterochromaffin Cells, Humans, Intestinal Mucosa, Serotonin},
pubstate = {published},
tppubtype = {article}
}
Enteroendocrine (EE) cells within the intestinal epithelium produce a range of hormones that have key roles in modulating satiety and feeding behavior in humans. The regulation of hormone release from EE cells as a potential therapeutic strategy to treat metabolic disorders is highly sought after by the pharmaceutical industry. However, functional studies are limited by the scarcity of EE cells (or surrogates) in both in vivo and in vitro systems. Enterochromaffin (EC) cells are a subtype of EE cells that produce serotonin (5HT). Here, we explored simple strategies to enrich EC cells in in vitro monolayer systems derived from human primary intestinal stem cells. During differentiation of the monolayers, the EC cell lineage was significantly altered by both the culture method [air-liquid interface (ALI) vs submerged] and the presence of vasoactive intestinal peptide (VIP). Compared with traditional submerged cultures without VIP, VIP-assisted ALI culture significantly boosted the number of EC cells and their 5HT secretion by up to 430 and 390%, respectively. The method also increased the numbers of other subtypes of EE cells such as L cells. Additionally, this method generated monolayers with enhanced barrier integrity, so that directional (basal or apical) 5HT secretion was measurable. For all donor tissues, the enriched EC cells improved the signal-to-background ratio and reliability of 5HT release assays. The enhancement in the 5HT secretion behavior was consistent over time from a single donor, but significant variation in the amount of secreted 5HT was present among tissues derived from five different donors. To demonstrate the utility of the EC-enriched monolayer system, 13 types of pungent food ingredients were screened for their ability to stimulate 5HT secretion. Curcumin found in the spice turmeric derived from the Curcuma longa plant was found to be the most potent secretagogue. This EC-enriched cell monolayer platform can provide a valuable analytical tool for the high-throughput screening of nutrients and gut microbial components that alter the secretion of 5HT.
Donowitz, Mark; Turner, Jerrold R.; Verkman, Alan S.; Zachos, Nicholas Constantine
Current and potential future applications of human stem cell models in drug development Journal Article
In: vol. 130, no. 7, pp. 3342–3344, 2020, ISSN: 0021-9738, (Publisher: American Society for Clinical Investigation).
@article{donowitz_current_2020,
title = {Current and potential future applications of human stem cell models in drug development},
author = {Mark Donowitz and Jerrold R. Turner and Alan S. Verkman and Nicholas Constantine Zachos},
url = {https://www.jci.org/articles/view/138645},
doi = {10.1172/JCI138645},
issn = {0021-9738},
year = {2020},
date = {2020-07-01},
urldate = {2023-03-03},
volume = {130},
number = {7},
pages = {3342–3344},
note = {Publisher: American Society for Clinical Investigation},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Gareb, Bahez; Otten, Antonius T.; Frijlink, Henderik W.; Dijkstra, Gerard; Kosterink, Jos G. W.
Review: Local Tumor Necrosis Factor-α Inhibition in Inflammatory Bowel Disease Journal Article
In: vol. 12, no. 6, pp. 539, 2020, ISSN: 1999-4923.
Abstract | Links | BibTeX | Tags:
@article{gareb_review_2020,
title = {Review: Local Tumor Necrosis Factor-α Inhibition in Inflammatory Bowel Disease},
author = {Bahez Gareb and Antonius T. Otten and Henderik W. Frijlink and Gerard Dijkstra and Jos G. W. Kosterink},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7356880/},
doi = {10.3390/pharmaceutics12060539},
issn = {1999-4923},
year = {2020},
date = {2020-06-11},
urldate = {2023-05-04},
volume = {12},
number = {6},
pages = {539},
abstract = {Crohn’s disease (CD) and ulcerative colitis (UC) are inflammatory bowel diseases (IBD) characterized by intestinal inflammation. Increased intestinal levels of the proinflammatory cytokine tumor necrosis factor-α (TNF-α) are associated with disease activity and severity. Anti-TNF-α therapy is administered systemically and efficacious in the treatment of IBD. However, systemic exposure is associated with adverse events that may impede therapeutic treatment. Clinical studies show that the efficacy correlates with immunological effects localized in the gastrointestinal tract (GIT) as opposed to systemic effects. These data suggest that site-specific TNF-α inhibition in IBD may be efficacious with fewer expected side effects related to systemic exposure. We therefore reviewed the available literature that investigated the efficacy or feasibility of local TNF-α inhibition in IBD. A literature search was performed on PubMed with given search terms and strategy. Of 8739 hits, 48 citations were included in this review. These studies ranged from animal studies to randomized placebo-controlled clinical trials. In these studies, local anti-TNF-α therapy was achieved with antibodies, antisense oligonucleotides (ASO), small interfering RNA (siRNA), microRNA (miRNA) and genetically modified organisms. This narrative review summarizes and discusses these approaches in view of the clinical relevance of local TNF-α inhibition in IBD.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Crohn’s disease (CD) and ulcerative colitis (UC) are inflammatory bowel diseases (IBD) characterized by intestinal inflammation. Increased intestinal levels of the proinflammatory cytokine tumor necrosis factor-α (TNF-α) are associated with disease activity and severity. Anti-TNF-α therapy is administered systemically and efficacious in the treatment of IBD. However, systemic exposure is associated with adverse events that may impede therapeutic treatment. Clinical studies show that the efficacy correlates with immunological effects localized in the gastrointestinal tract (GIT) as opposed to systemic effects. These data suggest that site-specific TNF-α inhibition in IBD may be efficacious with fewer expected side effects related to systemic exposure. We therefore reviewed the available literature that investigated the efficacy or feasibility of local TNF-α inhibition in IBD. A literature search was performed on PubMed with given search terms and strategy. Of 8739 hits, 48 citations were included in this review. These studies ranged from animal studies to randomized placebo-controlled clinical trials. In these studies, local anti-TNF-α therapy was achieved with antibodies, antisense oligonucleotides (ASO), small interfering RNA (siRNA), microRNA (miRNA) and genetically modified organisms. This narrative review summarizes and discusses these approaches in view of the clinical relevance of local TNF-α inhibition in IBD.
Koelink, Pim J.; Bloemendaal, Felicia M.; Li, Bofeng; Westera, Liset; Vogels, Esther W. M.; van Roest, Manon; Gloudemans, Anouk K.; van 't Wout, Angelique B.; Korf, Hannelie; Vermeire, Séverine; te Velde, Anje A.; Ponsioen, Cyriel Y.; D'Haens, Geert RAM; Verbeek, J. Sjef; Geiger, Terrence L.; Wildenberg, Manon E.; van den Brink, Gijs R.
Anti-TNF therapy in IBD exerts its therapeutic effect through macrophage IL-10 signalling Journal Article
In: vol. 69, no. 6, pp. 1053–1063, 2020, ISSN: 0017-5749, 1468-3288, (Publisher: BMJ Publishing Group Section: Inflammatory bowel disease).
Abstract | Links | BibTeX | Tags: {IBD} basic research, {TNF}, antibody targeted therapy, infliximab, interleukins
@article{koelink_anti-tnf_2020,
title = {Anti-TNF therapy in IBD exerts its therapeutic effect through macrophage IL-10 signalling},
author = {Pim J. Koelink and Felicia M. Bloemendaal and Bofeng Li and Liset Westera and Esther W. M. Vogels and Manon van Roest and Anouk K. Gloudemans and Angelique B. van 't Wout and Hannelie Korf and Séverine Vermeire and Anje A. te Velde and Cyriel Y. Ponsioen and Geert RAM D'Haens and J. Sjef Verbeek and Terrence L. Geiger and Manon E. Wildenberg and Gijs R. van den Brink},
url = {https://gut.bmj.com/content/69/6/1053},
doi = {10.1136/gutjnl-2019-318264},
issn = {0017-5749, 1468-3288},
year = {2020},
date = {2020-06-01},
urldate = {2023-05-03},
volume = {69},
number = {6},
pages = {1053–1063},
abstract = {Objective Macrophage interleukin (IL)-10 signalling plays a critical role in the maintenance of a regulatory phenotype that prevents the development of IBD. We have previously found that anti-tumour necrosis factor (TNF) monoclonal antibodies act through Fcγ-receptor (FcγR) signalling to promote repolarisation of proinflammatory intestinal macrophages to a CD206+ regulatory phenotype. The role of IL-10 in anti-TNF-induced macrophage repolarisation has not been examined.
Design We used human peripheral blood monocytes and mouse bone marrow-derived macrophages to study IL-10 production and CD206+ regulatory macrophage differentiation. To determine whether the efficacy of anti-TNF was dependent on IL-10 signalling in vivo and in which cell type, we used the CD4+CD45Rbhigh T-cell transfer model in combination with several genetic mouse models.
Results Anti-TNF therapy increased macrophage IL-10 production in an FcγR-dependent manner, which caused differentiation of macrophages to a more regulatory CD206+ phenotype in vitro. Pharmacological blockade of IL-10 signalling prevented the induction of these CD206+ regulatory macrophages and diminished the therapeutic efficacy of anti-TNF therapy in the CD4+CD45Rbhigh T-cell transfer model of IBD. Using cell type-specific IL-10 receptor mutant mice, we found that IL-10 signalling in macrophages but not T cells was critical for the induction of CD206+ regulatory macrophages and therapeutic response to anti-TNF.
Conclusion The therapeutic efficacy of anti-TNF in resolving intestinal inflammation is critically dependent on IL-10 signalling in macrophages.},
note = {Publisher: BMJ Publishing Group
Section: Inflammatory bowel disease},
keywords = {{IBD} basic research, {TNF}, antibody targeted therapy, infliximab, interleukins},
pubstate = {published},
tppubtype = {article}
}
Objective Macrophage interleukin (IL)-10 signalling plays a critical role in the maintenance of a regulatory phenotype that prevents the development of IBD. We have previously found that anti-tumour necrosis factor (TNF) monoclonal antibodies act through Fcγ-receptor (FcγR) signalling to promote repolarisation of proinflammatory intestinal macrophages to a CD206+ regulatory phenotype. The role of IL-10 in anti-TNF-induced macrophage repolarisation has not been examined.
Design We used human peripheral blood monocytes and mouse bone marrow-derived macrophages to study IL-10 production and CD206+ regulatory macrophage differentiation. To determine whether the efficacy of anti-TNF was dependent on IL-10 signalling in vivo and in which cell type, we used the CD4+CD45Rbhigh T-cell transfer model in combination with several genetic mouse models.
Results Anti-TNF therapy increased macrophage IL-10 production in an FcγR-dependent manner, which caused differentiation of macrophages to a more regulatory CD206+ phenotype in vitro. Pharmacological blockade of IL-10 signalling prevented the induction of these CD206+ regulatory macrophages and diminished the therapeutic efficacy of anti-TNF therapy in the CD4+CD45Rbhigh T-cell transfer model of IBD. Using cell type-specific IL-10 receptor mutant mice, we found that IL-10 signalling in macrophages but not T cells was critical for the induction of CD206+ regulatory macrophages and therapeutic response to anti-TNF.
Conclusion The therapeutic efficacy of anti-TNF in resolving intestinal inflammation is critically dependent on IL-10 signalling in macrophages.
Design We used human peripheral blood monocytes and mouse bone marrow-derived macrophages to study IL-10 production and CD206+ regulatory macrophage differentiation. To determine whether the efficacy of anti-TNF was dependent on IL-10 signalling in vivo and in which cell type, we used the CD4+CD45Rbhigh T-cell transfer model in combination with several genetic mouse models.
Results Anti-TNF therapy increased macrophage IL-10 production in an FcγR-dependent manner, which caused differentiation of macrophages to a more regulatory CD206+ phenotype in vitro. Pharmacological blockade of IL-10 signalling prevented the induction of these CD206+ regulatory macrophages and diminished the therapeutic efficacy of anti-TNF therapy in the CD4+CD45Rbhigh T-cell transfer model of IBD. Using cell type-specific IL-10 receptor mutant mice, we found that IL-10 signalling in macrophages but not T cells was critical for the induction of CD206+ regulatory macrophages and therapeutic response to anti-TNF.
Conclusion The therapeutic efficacy of anti-TNF in resolving intestinal inflammation is critically dependent on IL-10 signalling in macrophages.
Ananthakrishnan, Ashwin N.; Kaplan, Gilaad G.; Ng, Siew C.
Changing Global Epidemiology of Inflammatory Bowel Diseases: Sustaining Health Care Delivery Into the 21st Century Journal Article
In: vol. 18, no. 6, pp. 1252–1260, 2020, ISSN: 1542-3565, 1542-7714, (Publisher: Elsevier).
Links | BibTeX | Tags: {CD}, {FDR}, {IBD}, {UC}, Crohn’s disease, Crohn's disease, first-degree relative, inflammatory bowel disease, Ulcerative Colitis
@article{ananthakrishnan_changing_2020,
title = {Changing Global Epidemiology of Inflammatory Bowel Diseases: Sustaining Health Care Delivery Into the 21st Century},
author = {Ashwin N. Ananthakrishnan and Gilaad G. Kaplan and Siew C. Ng},
url = {https://www.cghjournal.org/article/S1542-3565%2820%2930107-5/fulltext},
doi = {10.1016/j.cgh.2020.01.028},
issn = {1542-3565, 1542-7714},
year = {2020},
date = {2020-05-01},
urldate = {2023-04-14},
volume = {18},
number = {6},
pages = {1252–1260},
note = {Publisher: Elsevier},
keywords = {{CD}, {FDR}, {IBD}, {UC}, Crohn’s disease, Crohn's disease, first-degree relative, inflammatory bowel disease, Ulcerative Colitis},
pubstate = {published},
tppubtype = {article}
}