title: Retinoblastoma protein controls growth, survival and neuronal migration in human cerebral organoids.
abstract: The tumor suppressor retinoblastoma protein (RB) regulates S-phase cell cycle entry via E2F transcription factors. Knockout (KO) mice have shown that RB plays roles in cell migration, differentiation and apoptosis, in developing and adult brain. In addition, the RB family is required for self-renewal and survival of human embryonic stem cells (hESCs). Since little is known about the role of RB in human brain development, we investigated its function in cerebral organoids differentiated from gene-edited hESCs lacking RB. We show that RB is abundantly expressed in neural stem and progenitor cells in organoids at 15 and 28 days of culture. RB loss promoted S-phase entry in DCX+cells and increased apoptosis in Sox2+neural stem and progenitor cells, and in DCX+and Tuj1+neurons. Associated with these cell cycle and pro-apoptotic effects, we observed increasedCCNA2andBAXgene expression, respectively. Moreover, we observed aberrant Tuj1+neuronal migration in RB-KO organoids and upregulation of the gene encoding VLDLR, a receptor important in reelin signaling. Corroborating the results in RB-KO organoidsin vitro, we observed ectopically localized Tuj1+cells in RB-KO teratomas grownin vivoTaken together, these results identify crucial functions for RB in the cerebral organoid model of human brain development.
authors.affiliation: Department of Molecular Biology and Hamon Center for Regenerative Science and Medicine, UT Southwestern Medical Center, Dallas, TX 75390, USA.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Matsui
authors.affiliation: Department of Molecular Biology and Hamon Center for Regenerative Science and Medicine, UT Southwestern Medical Center, Dallas, TX 75390, USA.
authors.forename: Vanesa
authors.initials: V
authors.lastname: Nieto-Estévez
authors.affiliation: Department of Internal Medicine and Hamon Center for Regenerative Science and Medicine, UT Southwestern Medical Center, Dallas, TX 75390, USA.
authors.forename: Sergii
authors.initials: S
authors.lastname: Kyrychenko
authors.affiliation: Department of Internal Medicine and Hamon Center for Regenerative Science and Medicine, UT Southwestern Medical Center, Dallas, TX 75390, USA jenny.hsieh@utsouthwestern.edu jay.schneider@utsouthwestern.edu.
authors.forename: Jay W
authors.initials: JW
authors.lastname: Schneider
authors.affiliation: Department of Molecular Biology and Hamon Center for Regenerative Science and Medicine, UT Southwestern Medical Center, Dallas, TX 75390, USA jenny.hsieh@utsouthwestern.edu jay.schneider@utsouthwestern.edu.
authors.forename: Jenny
authors.identifier: 0000-0003-0214-0771
authors.initials: J
authors.lastname: Hsieh
mesh_terms: D016923:Cell Death; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D054022:Cerebrum / Q000166:cytology*; D000089662:Doublecortin Protein; D053595:Embryonic Stem Cells / Q000166:cytology; D017353:Gene Deletion; D055786:Gene Knockout Techniques; D006801:Humans; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D009940:Organoids / Q000166:cytology* / Q000378:metabolism*; D000091042:Reelin Protein; D016160:Retinoblastoma Protein / Q000378:metabolism*; D016196:S Phase
chemical_list: C000714847:Dcx protein, mouse; D000089662:Doublecortin Protein; D000091042:Reelin Protein; D016160:Retinoblastoma Protein; C000717127:RELN protein, human; C000717128:Reln protein, mouse
keywords: Brain; CRISPR/Cas9; Cell cycle; Human embryonic stem cells; Organoid; Retinoblastoma