title: Activation of RhoA-ROCK-BMP signaling reprograms adult human corneal endothelial cells.
abstract: Currently there are limited treatment options for corneal blindness caused by dysfunctional corneal endothelial cells. The primary treatment involves transplantation of healthy donor human corneal endothelial cells, but a global shortage of donor corneas necessitates other options. Conventional tissue approaches for corneal endothelial cells are based on EDTA-trypsin treatment and run the risk of irreversible endothelial mesenchymal transition by activating canonical Wingless-related integration site (Wnt) and TGF-β signaling. Herein, we demonstrate an alternative strategy that avoids disruption of cell-cell junctions and instead activates Ras homologue gene family A (RhoA)-Rho-associated protein kinase (ROCK)-canonical bone morphogenic protein signaling to reprogram adult human corneal endothelial cells to neural crest-like progenitors via activation of the miR302b-Oct4-Sox2-Nanog network. This approach allowed us to engineer eight human corneal endothelial monolayers of transplantable size, with a normal density and phenotype from one corneoscleral rim. Given that a similar signal network also exists in the retinal pigment epithelium, this partial reprogramming approach may have widespread relevance and potential for treating degenerative diseases.
authors.affiliation: TissueTech, Inc., Ocular Surface Center, and Ocular Surface Research & Education Foundation, Miami, FL 33173.
authors.forename: Ying-Ting
authors.initials: YT
authors.lastname: Zhu
authors.affiliation: Pediatric Research Institute and Department of Pediatric Hematology, Qilu Children's Hospital, Shandong University, Jinan, Shandong 250022, People's Republic of China.
authors.forename: Fu
authors.initials: F
authors.lastname: Li
authors.affiliation: Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, People's Republic of China.
authors.forename: Bo
authors.initials: B
authors.lastname: Han
authors.affiliation: TissueTech, Inc., Ocular Surface Center, and Ocular Surface Research & Education Foundation, Miami, FL 33173.
authors.forename: Sean
authors.initials: S
authors.lastname: Tighe
authors.affiliation: TissueTech, Inc., Ocular Surface Center, and Ocular Surface Research & Education Foundation, Miami, FL 33173.
authors.forename: Suzhen
authors.initials: S
authors.lastname: Zhang
authors.affiliation: TissueTech, Inc., Ocular Surface Center, and Ocular Surface Research & Education Foundation, Miami, FL 33173.
authors.forename: Szu-Yu
authors.initials: SY
authors.lastname: Chen
authors.affiliation: Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: TissueTech, Inc., Ocular Surface Center, and Ocular Surface Research & Education Foundation, Miami, FL 33173 stseng@ocularsurface.com.
authors.forename: Scheffer C G
authors.initials: SC
authors.lastname: Tseng
mesh_terms: D052004:Bone Morphogenetic Protein Receptors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D003315:Cornea / Q000166:cytology* / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D018398:Homeodomain Proteins; D006801:Humans; D035683:MicroRNAs; D000071317:Nanog Homeobox Protein; D009432:Neural Crest / Q000166:cytology; D050814:Octamer Transcription Factor-3; D034622:RNA Interference; D034741:RNA, Small Interfering; D055748:SOXB1 Transcription Factors; D013234:Stem Cells; D051996:Transcription Factor RelA / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D060449:Wnt Signaling Pathway; D020729:p120 GTPase Activating Protein / Q000235:genetics; D054460:rho-Associated Kinases / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D018398:Homeodomain Proteins; C532169:MIRN302A microRNA, human; D035683:MicroRNAs; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D034741:RNA, Small Interfering; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D051996:Transcription Factor RelA; D014157:Transcription Factors; C492254:ZBTB33 protein, human; D020729:p120 GTPase Activating Protein; C066737:RHOA protein, human; C515851:ROCK1 protein, human; D054460:rho-Associated Kinases; D052004:Bone Morphogenetic Protein Receptors; D020742:rhoA GTP-Binding Protein