title: Nuclear p120 catenin unlocks mitotic block of contact-inhibited human corneal endothelial monolayers without disrupting adherent junctions.
abstract: Contact inhibition ubiquitously exists in non-transformed cells that are in contact with neighboring cells. This phenomenon explains the poor regenerative capacity of in vivo human corneal endothelial cells during aging, injury and surgery. This study demonstrated that the conventional approach of expanding human corneal endothelial cells by disrupting contact inhibition with EDTA followed by bFGF activated canonical Wnt signaling and lost the normal phenotype to endothelial-mesenchymal transition, especially if TGFβ1 was added. By contrast, siRNA against p120 catenin (CTNND1) also uniquely promoted proliferation of the endothelial cells by activating trafficking of p120 catenin to the nucleus, thus relieving repression by nuclear Kaiso. This nuclear p120-catenin-Kaiso signaling is associated with activation of RhoA-ROCK signaling, destabilization of microtubules and inhibition of Hippo signaling, but not with activation of Wnt-β-catenin signaling. Consequently, proliferating human corneal endothelial cells maintained a hexagonal shape, with junctional expression of N-cadherin, ZO-1 and Na(+)/K(+)-ATPase. Further expansion of human corneal endothelial monolayers with a normal phenotype and a higher density was possible by prolonging treatment with p120 catenin siRNA followed by its withdrawal. This new strategy of perturbing contact inhibition by selective activation of p120-catenin-Kaiso signaling without disrupting adherent junction could be used to engineer surgical grafts containing normal human corneal endothelial cells to meet a global corneal shortage and for endothelial keratoplasties.
authors.affiliation: Research and Development Department, TissueTech Inc, Ocular Surface Center and Ocular Surface Research and Education Foundation, Miami, FL 33173, USA.
authors.forename: Ying-Ting
authors.initials: YT
authors.lastname: Zhu
authors.forename: Hung-Chi
authors.initials: HC
authors.lastname: Chen
authors.forename: Szu-Yu
authors.initials: SY
authors.lastname: Chen
authors.forename: Scheffer C G
authors.initials: SC
authors.lastname: Tseng
mesh_terms: D022005:Adherens Junctions / Q000187:drug effects / Q000502:physiology*; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D051177:Catenins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D048708:Cell Growth Processes / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003260:Contact Inhibition / Q000187:drug effects / Q000502:physiology*; D003315:Cornea / Q000166:cytology* / Q000187:drug effects; D004492:Edetic Acid / Q000494:pharmacology; D004728:Endothelium, Corneal / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D008875:Middle Aged; D008938:Mitosis / Q000187:drug effects / Q000502:physiology*; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D051153:Wnt Proteins / Q000378:metabolism; D055815:Young Adult; D054460:rho-Associated Kinases / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism; D000095002:Delta Catenin
chemical_list: D051177:Catenins; D034741:RNA, Small Interfering; D014157:Transcription Factors; D053773:Transforming Growth Factor beta1; D051153:Wnt Proteins; C492254:ZBTB33 protein, human; D016222:Fibroblast Growth Factor 2; C066737:RHOA protein, human; D004492:Edetic Acid; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein; D000095002:Delta Catenin; C000722347:CTNND1 protein, human