Modulation of integrin and E-cadherin-mediated adhesions to spatially control heterogeneity in human pluripotent stem cell differentiation.
Heterogeneity in human pluripotent stem cell (PSC) fates is partially caused by mechanical asymmetry arising from spatial polarization of cell-cell and cell-matrix adhesions. Independent studies have shown that integrin and E-cadherin adhesions promote opposing differentiation and pluripotent fates respectively although their crosstalk mechanism in modulating cell fate heterogeneity remains unknown. Here, we demonstrated that spatial polarization of integrin and E-cadherin adhesions in a human PSC colony compete to recruit Rho-ROCK activated myosin II to different localities to pattern pluripotent-differentiation decisions, resulting in spatially heterogeneous colonies. Cell micropatterning was used to modulate the spatial polarization of cell adhesions, which enabled us to prospectively determine localization patterns of activated myosin II and mesoendoderm differentiation. Direct inhibition of Rho-ROCK-myosin II activation phenocopied E-cadherin rather than integrin inhibition to form uniformly differentiated colonies. This indicated that E-cadherin was the primary gatekeeper to differentiation progression. This insight allows for biomaterials to be tailored for human PSC maintenance or differentiation with minimal heterogeneity.
Institute of Bioengineering and Nanotechnology, A*STAR, #04-01, 31 Biopolis Way, 138669 Singapore, Singapore; Department of Biomedical Engineering, National University of Singapore, 9 Engineering Drive 1, EA #03-12, Singapore 117575, Singapore. Electronic address: biety@nus.edu.sg.
Yi-Chin
YC
Toh
Institute of Bioengineering and Nanotechnology, A*STAR, #04-01, 31 Biopolis Way, 138669 Singapore, Singapore; Mechanobiology Institute, National University of Singapore, T-Lab, #10-01, 5A Engineering Drive 1, Singapore 117411, Singapore.
Jiangwa
J
Xing
Institute of Bioengineering and Nanotechnology, A*STAR, #04-01, 31 Biopolis Way, 138669 Singapore, Singapore; Mechanobiology Institute, National University of Singapore, T-Lab, #10-01, 5A Engineering Drive 1, Singapore 117411, Singapore; Department of Physiology, Yong Loo Lin School of Medicine, National University Health System, MD9 #04-11, 2 Medical Drive, Singapore 117597, Singapore; NUS Graduate School for Integrative Sciences and Engineering, Centre for Life Sciences (CeLS), #05-01, 28 Medical Drive, Singapore 117456, Singapore; Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing; #B-10, Singapore 138602, Singapore; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA. Electronic address: hanry_yu@nuhs.edu.sg.
Hanry
H
Yu
D000205:Actomyosin / Q000378:metabolism; D015703:Antigens, CD; D015820:Cadherins / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D016764:Cell Polarity / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004707:Endoderm / Q000166:cytology / Q000187:drug effects; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D007797:Laminin / Q000494:pharmacology; D008648:Mesoderm / Q000166:cytology / Q000187:drug effects; D024462:Myosin Type II / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D011509:Proteoglycans / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D054460:rho-Associated Kinases / Q000378:metabolism
D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; D004338:Drug Combinations; D016023:Integrins; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D000205:Actomyosin; D054460:rho-Associated Kinases; D024462:Myosin Type II
Cell fate heterogeneity; Human pluripotent stem cells; Mechanical forces; Micropatterning