Inducers of the endothelial cell barrier identified through chemogenomic screening in genome-edited hPSC-endothelial cells
| العنوان: | Inducers of the endothelial cell barrier identified through chemogenomic screening in genome-edited hPSC-endothelial cells |
|---|---|
| المؤلفون: | Leonard D. Goldstein, Filip Roudnicky, Bo Kyoung Kim, Claas A. Meyer, Lisa Sach-Peltason, Nikhil J. Pandya, Klaus Christensen, Christoph Patsch, Héloïse Ragelle, Oliv Eidam, Chad A. Cowan, Martin Graf, Pamela Strassburger, Zora Modrusan, Mark Burcin, Balazs Banfai, Verena Kueppers, Manuel Tzouros, Mirjana Lazendic, Sabine Uhles, Gregor Sturm, Peter D. Westenskow, Franco Revelant, Yanjun Lan, Jitao David Zhang, Sabine Gruener |
| المصدر: | Proc Natl Acad Sci U S A |
| بيانات النشر: | Proceedings of the National Academy of Sciences, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Pluripotent Stem Cells, Vascular Endothelial Growth Factor A, Pyridines, Drug Evaluation, Preclinical, Cell Line, Tight Junctions, Small Molecule Libraries, Transcriptome, Mice, Blood-Retinal Barrier, Animals, Humans, Kinome, Claudin-5, Induced pluripotent stem cell, Cell Proliferation, Gene Editing, Mice, Knockout, Genome, Multidisciplinary, SI Corrections, Tight junction, Chemistry, Wnt signaling pathway, Endothelial Cells, Cell Differentiation, Biological Sciences, Cell biology, Endothelial stem cell, Vascular endothelial growth factor A, Blood-Brain Barrier, Paracellular transport, Pyrazoles |
| الوصف: | The blood–retina barrier and blood–brain barrier (BRB/BBB) are selective and semipermeable and are critical for supporting and protecting central nervous system (CNS)-resident cells. Endothelial cells (ECs) within the BRB/BBB are tightly coupled, express high levels of Claudin-5 (CLDN5), a junctional protein that stabilizes ECs, and are important for proper neuronal function. To identify novel CLDN5 regulators (and ultimately EC stabilizers), we generated a CLDN5-P2A-GFP stable cell line from human pluripotent stem cells (hPSCs), directed their differentiation to ECs (CLDN5-GFP hPSC-ECs), and performed flow cytometry-based chemogenomic library screening to measure GFP expression as a surrogate reporter of barrier integrity. Using this approach, we identified 62 unique compounds that activated CLDN5-GFP. Among them were TGF-β pathway inhibitors, including RepSox. When applied to hPSC-ECs, primary brain ECs, and retinal ECs, RepSox strongly elevated barrier resistance (transendothelial electrical resistance), reduced paracellular permeability (fluorescein isothiocyanate-dextran), and prevented vascular endothelial growth factor A (VEGFA)-induced barrier breakdown in vitro. RepSox also altered vascular patterning in the mouse retina during development when delivered exogenously. To determine the mechanism of action of RepSox, we performed kinome-, transcriptome-, and proteome-profiling and discovered that RepSox inhibited TGF-β, VEGFA, and inflammatory gene networks. In addition, RepSox not only activated vascular-stabilizing and barrier-establishing Notch and Wnt pathways, but also induced expression of important tight junctions and transporters. Taken together, our data suggest that inhibiting multiple pathways by selected individual small molecules, such as RepSox, may be an effective strategy for the development of better BRB/BBB models and novel EC barrier-inducing therapeutics. |
| تدمد: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1911532117 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2681eba3f4ec57913642788bbe14e23c https://doi.org/10.1073/pnas.1911532117 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....2681eba3f4ec57913642788bbe14e23c |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10916490 00278424 |
|---|---|
| DOI: | 10.1073/pnas.1911532117 |