1. Role of RB1 in human embryonic stem cell-derived retinal organoids.
- Author
-
Zheng C, Schneider JW, and Hsieh J
- Subjects
- Animals, Apoptosis physiology, CRISPR-Cas Systems, Cell Differentiation genetics, Human Embryonic Stem Cells cytology, Humans, Male, Mice, Mice, Inbred NOD, Mice, SCID, Organoids cytology, Pluripotent Stem Cells cytology, Retina physiology, Retinal Ganglion Cells metabolism, Retinal Neoplasms metabolism, Retinoblastoma metabolism, Retinoblastoma Binding Proteins physiology, Ubiquitin-Protein Ligases physiology, Human Embryonic Stem Cells metabolism, Retina embryology, Retinoblastoma Binding Proteins metabolism, Ubiquitin-Protein Ligases metabolism
- Abstract
Three-dimensional (3D) organoid models derived from human pluripotent stem cells provide a platform for studying human development and understanding disease mechanisms. Most studies that examine biallelic inactivation of the cell cycle regulator Retinoblastoma 1 (RB1) and the link to retinoblastoma is in mice, however, less is known regarding the pathophysiological role of RB1 during human retinal development. To study the role of RB1 in early human retinal development and tumor formation, we generated retinal organoids from CRISPR/Cas9-derived RB1-null human embryonic stem cells (hESCs). We showed that RB is abundantly expressed in retinal progenitor cells in retinal organoids and loss of RB1 promotes S-phase entry. Furthermore, loss of RB1 resulted in widespread apoptosis and reduced the number of photoreceptor, ganglion, and bipolar cells. Interestingly, RB1 mutation in retinal organoids did not result in retinoblastoma formation in vitro or in the vitreous body of NOD/SCID immunodeficient mice. Together, our work identifies a crucial function for RB1 in human retinal development and suggests that RB1 deletion alone is not sufficient for tumor development, at least in human retinal organoids., Competing Interests: Declaration of competing of interest The authors indicate no potential conflicts of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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