Publications
Browse peer-reviewed literature, posters, webinars, blog articles, and more showing how we and others are using RepliGut Systems to support discovery.
2024
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: Lab Chip, vol. 24, no. 2, pp. 339–355, 2024, ISSN: 1473-0189, (Publisher: The Royal Society of Chemistry).
Abstract | Links | BibTeX | Tags: absorption, Bioavailability, Biological Transport, Caco-2 Cells, drug absorption, drug metabolising enzymes ({DME}), drug permeability, intestinal barrier, oral bioavailability, permeability
@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 = {2024-01-17},
journal = {Lab Chip},
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 = {absorption, Bioavailability, Biological Transport, Caco-2 Cells, drug absorption, drug metabolising enzymes ({DME}), drug permeability, intestinal barrier, oral bioavailability, permeability},
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.
2018
Gunasekara, Dulan B.; Speer, Jennifer; Wang, Yuli; Nguyen, Daniel L.; Reed, Mark I.; Smiddy, Nicole M.; Parker, Joel S.; Fallon, John K.; Smith, Philip C.; Sims, Christopher E.; Magness, Scott T.; Allbritton, Nancy L.
A monolayer of primary colonic epithelium generated on a scaffold with a gradient of stiffness for drug transport studies Journal Article
In: vol. 90, no. 22, pp. 13331, 2018, (Publisher: NIH Public Access).
Abstract | Links | BibTeX | Tags: Atenolol, Biological Transport, Caco-2 Cells, Colon, Epithelium, Humans, Propranolol, Riboflavin, Tissue Engineering
@article{gunasekara_monolayer_2018,
title = {A monolayer of primary colonic epithelium generated on a scaffold with a gradient of stiffness for drug transport studies},
author = {Dulan B. Gunasekara and Jennifer Speer and Yuli Wang and Daniel L. Nguyen and Mark I. Reed and Nicole M. Smiddy and Joel S. Parker and John K. Fallon and Philip C. Smith and Christopher E. Sims and Scott T. Magness and Nancy L. Allbritton},
url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6339567/},
doi = {10.1021/acs.analchem.8b02845},
year = {2018},
date = {2018-11-11},
urldate = {2018-11-11},
volume = {90},
number = {22},
pages = {13331},
abstract = {Animal models are frequently used for in vitro physiologic and drug transport studies of the colon, but there exists significant pressure to improve assay throughput as well as achieve tighter control of experimental variables than can be achieved with ...},
note = {Publisher: NIH Public Access},
keywords = {Atenolol, Biological Transport, Caco-2 Cells, Colon, Epithelium, Humans, Propranolol, Riboflavin, Tissue Engineering},
pubstate = {published},
tppubtype = {article}
}
Animal models are frequently used for in vitro physiologic and drug transport studies of the colon, but there exists significant pressure to improve assay throughput as well as achieve tighter control of experimental variables than can be achieved with ...