Single-Cell RNA Sequencing of hESC-Derived 3D Retinal Organoids Reveals Novel Genes Regulating RPC Commitment in Early Human Retinogenesis.
Xiying Mao, Qin An, Huiyu Xi, Xian-Jie Yang, Xiangmei Zhang, Songtao Yuan, Jinmei Wang, Youjin Hu, Qinghuai Liu, Guoping Fan
Author Information
Xiying Mao: Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
Qin An: Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, 695 Charles Young Drive South, Los Angeles, CA 90095, USA.
Huiyu Xi: Department of Ophthalmology, Xuzhou First People's Hospital of Xuzhou Medical University, Xuzhou Eye Research Institute, Xuzhou 221002, China.
Xian-Jie Yang: Stein Eye Institute, Molecular Biology Institute, University of California, Los Angeles, CA 90095, USA.
Xiangmei Zhang: Stein Eye Institute, Molecular Biology Institute, University of California, Los Angeles, CA 90095, USA.
Songtao Yuan: Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
Jinmei Wang: Institute of Regenerative Medicine and International Lab of Ocular Stem Cells at Shanghai East Hospital, School of Life Science &Technology, Tongji University, Shanghai 200092, China.
Youjin Hu: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China. Electronic address: huyoujin@gzzoc.com.
Qinghuai Liu: Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. Electronic address: liuqh@njmu.edu.cn.
Guoping Fan: Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, 695 Charles Young Drive South, Los Angeles, CA 90095, USA. Electronic address: gfan@mednet.ucla.edu.
The development of the mammalian retina is a complicated process involving the generation of distinct types of neurons from retinal progenitor cells (RPCs) in a spatiotemporal-specific manner. The progression of RPCs during retinogenesis includes RPC proliferation, cell-fate commitment, and specific neuronal differentiation. In this study, by performing single-cell RNA sequencing of cells isolated from human embryonic stem cell (hESC)-derived 3D retinal organoids, we successfully deconstructed the temporal progression of RPCs during early human retinogenesis. We identified two distinctive subtypes of RPCs with unique molecular profiles, namely multipotent RPCs and neurogenic RPCs. We found that genes related to the Notch and Wnt signaling pathways, as well as chromatin remodeling, were dynamically regulated during RPC commitment. Interestingly, our analysis identified that CCND1, a G-phase cell-cycle regulator, was coexpressed with ASCL1 in a cell-cycle-independent manner. Temporally controlled overexpression of CCND1 in retinal organoids demonstrated a role for CCND1 in promoting early retinal neurogenesis. Together, our results revealed critical pathways and novel genes in early retinogenesis of humans.
