title: Substrate stiffness orchestrates epithelial cellular heterogeneity with controlled proliferative pattern via E-cadherin/β-catenin mechanotransduction.
abstract: Epithelial cellular heterogeneity has been observed in pathological tissues with abnormal matrix stiffness and cells cultured on rigid substrates. However, it remains unclear how matrix stiffness influences cellular heterogeneity formation in multi-cellular population. Here, we demonstrated that cellular heterogeneity regulated by substrate stiffness is evident starting from the initial single-cell stage (indicated by cellular Young's modulus and morphology) until the resulting multi-cellular stage (indicated by cellular functions) through distinguished proliferative patterns. Epithelial cells on soft substrate proliferated in a neighbor-dependent manner with stronger E-cadherin expression and more homogeneous E-cadherin/β-catenin localization compared to those on coverslips, which resulted in reduced heterogeneity in downstream cellular functions of the multi-cellular population. In particular, decreased heterogeneity in human embryonic stem cells upon expansion and endodermal induction was achieved on soft substrate. Overall, our work provides new insights on mechanotransduction during epithelial proliferation which regulates the formation of cellular heterogeneity and potentially provides a highly efficient approach to regulate stem cell fate by fine-tuning substrate stiffness.
  
  
  
  This study demonstrates that cellular heterogeneity regulated by substrate stiffness is evident starting from the initial single-cell stage until the resulting multi-cellular stage through distinguished proliferative patterns. During this process, E-cadherin/β-catenin mechanotransduction is found to play important role in substrate stiffness-regulated epithelial cellular heterogeneity formation. In particular, decreased heterogeneity in human embryonic stem cells upon expansion and endodermal induction is achieved on soft substrate. Hence, we believe that this work not only provides new insights on mechanotransduction of E-cadherin/β-catenin which regulates the formation of cellular heterogeneity during proliferation, but also potentially provides a highly efficient approach to regulate stem cell fate by fine-tuning substrate stiffness.
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China; Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing 100084, China.
authors.forename: Bingjie
authors.initials: B
authors.lastname: Wang
authors.affiliation: School of Life Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Qin
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China; Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing 100084, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Department of Basic Medical Sciences, School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Tie
authors.initials: T
authors.lastname: Xia
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing 100084, China.
authors.forename: Jingyu
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China; Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing 100084, China.
authors.forename: Longwei
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing 100084, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Department of Plastic, Reconstructive and Aesthetic Surgery, Beijing Tsinghua Changgung Hospital, Medical Center, Tsinghua University, Beijing 102218, China; Hepato-biliary Surgery of the First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xu
authors.affiliation: Department of Plastic, Reconstructive and Aesthetic Surgery, Beijing Tsinghua Changgung Hospital, Medical Center, Tsinghua University, Beijing 102218, China.
authors.forename: Chenyu
authors.initials: C
authors.lastname: Huang
authors.affiliation: Department of Basic Medical Sciences, School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing 100084, China; China Orthopedic Regenerative Medicine Group, China. Electronic address: duyanan@tsinghua.edu.cn.
authors.forename: Yanan
authors.initials: Y
authors.lastname: Du
mesh_terms: D015703:Antigens, CD; D001696:Biomechanical Phenomena / Q000187:drug effects; D064113:CRISPR-Cas Systems / Q000187:drug effects; D015820:Cadherins / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D055119:Elastic Modulus / Q000187:drug effects; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D022001:Focal Adhesions / Q000187:drug effects / Q000378:metabolism; D055786:Gene Knockout Techniques; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D020100:Hydrogels / Q000494:pharmacology; D008099:Liver / Q000187:drug effects / Q000473:pathology; D040542:Mechanotransduction, Cellular / Q000187:drug effects*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D051176:beta Catenin / Q000378:metabolism*
chemical_list: D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; D020100:Hydrogels; D051176:beta Catenin; C437167:poly(ethylene glycol)diacrylate; D011092:Polyethylene Glycols
keywords: E-cadherin/β-catenin mechanotransduction; Epithelial cellular heterogeneity; Proliferative pattern; Substrate stiffness