Role of RB1 in human embryonic stem cell-derived retinal organoids.

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.

• Authors: Canbin Zheng, Jay W Schneider, Jenny Hsieh
• Journal: Developmental biology (Dev Biol) Vol. 462 Issue 2 Pg. 197-207 (06 15 2020) ISSN: 1095-564X [Electronic] United States
• PMID: 32197890 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2020 Elsevier Inc. All rights reserved.

Chemical References

• RB1 protein, human
• Retinoblastoma Binding Proteins
• Ubiquitin-Protein Ligases

Topics

• Animals
• Apoptosis (physiology)
• CRISPR-Cas Systems
• Cell Differentiation (genetics)
• Human Embryonic Stem Cells (cytology, metabolism)
• Humans
• Male
• Mice
• Mice, Inbred NOD
• Mice, SCID
• Organoids (cytology)
• Pluripotent Stem Cells (cytology)
• Retina (embryology, physiology)
• Retinal Ganglion Cells (metabolism)
• Retinal Neoplasms (metabolism)
• Retinoblastoma (metabolism)
• Retinoblastoma Binding Proteins (metabolism, physiology)
• Ubiquitin-Protein Ligases (metabolism, physiology)