title: Pluripotent stem cell model reveals essential roles for miR-450b-5p and miR-184 in embryonic corneal lineage specification.
abstract: Approximately 6 million people worldwide are suffering from severe visual impairments or blindness due to corneal diseases. Corneal allogeneic transplantation is often required to restore vision; however, shortage in corneal grafts and immunorejections remain major challenges. The molecular basis of corneal diseases is poorly understood largely due to lack of appropriate cellular models. Here, we described a robust differentiation of human-induced pluripotent stem cells (hiPSCs) derived from hair follicles or skin fibroblasts into corneal epithelial-like cells. We found that BMP4, coupled with corneal fibroblast-derived conditioned medium and collagen IV allowed efficient corneal epithelial commitment of hiPSCs in a manner that recapitulated corneal epithelial lineage development with high purity. Organotypic reconstitution assays suggested the ability of these cells to stratify into a corneal-like epithelium. This model allowed us identifying miR-450b-5p as a molecular switch of Pax6, a major regulator of eye development. miR-450b-5p and Pax6 were reciprocally distributed at the presumptive epidermis and ocular surface, respectively. miR-450b-5p inhibited Pax6 expression and corneal epithelial fate in vitro, altogether, suggesting that by repressing Pax6, miR-450b-5p triggers epidermal specification of the ectoderm, while its absence allows ocular epithelial development. Additionally, miR-184 was detectable in early eye development and corneal epithelial differentiation of hiPSCs. The knockdown of miR-184 resulted in a decrease in Pax6 and K3, in line with recent findings showing that a point mutation in miR-184 leads to corneal dystrophy. Altogether, these data indicate that hiPSCs are valuable for modeling corneal development and may pave the way for future cell-based therapy.
authors.affiliation: INSERM U898, Nice, France. shalomfe@unice.fr
authors.forename: Ruby
authors.initials: R
authors.lastname: Shalom-Feuerstein
authors.forename: Laura
authors.initials: L
authors.lastname: Serror
authors.forename: Stephanie
authors.initials: S
authors.lastname: De La Forest Divonne
authors.forename: Isabelle
authors.initials: I
authors.lastname: Petit
authors.forename: Edith
authors.initials: E
authors.lastname: Aberdam
authors.forename: Livia
authors.initials: L
authors.lastname: Camargo
authors.forename: Odile
authors.initials: O
authors.lastname: Damour
authors.forename: Clotilde
authors.initials: C
authors.lastname: Vigouroux
authors.forename: Abraham
authors.initials: A
authors.lastname: Solomon
authors.forename: Cédric
authors.initials: C
authors.lastname: Gaggioli
authors.forename: Joseph
authors.initials: J
authors.lastname: Itskovitz-Eldor
authors.forename: Sajjad
authors.initials: S
authors.lastname: Ahmad
authors.forename: Daniel
authors.initials: D
authors.lastname: Aberdam
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage / Q000502:physiology*; D003315:Cornea / Q000166:cytology / Q000196:embryology*; D005136:Eye Proteins / Q000096:biosynthesis / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D018398:Homeodomain Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D008954:Models, Biological; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D005136:Eye Proteins; D018398:Homeodomain Proteins; C552233:MIRN184 microRNA, mouse; D035683:MicroRNAs; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; D012097:Repressor Proteins