Molecular analysis of RAX2-regulated retinal development using human retinal organoids at a single-cell resolution

Human embryonic stem cells (hESC)-derived retinal organoids are sophisticated in vitro systems for dissecting the complex dynamics of human retinal development. The formation of the human retina is a precisely organized process that depends on the regulated differentiation of retinal progenitor cell...

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Main Authors: Shaojun Wang, Yi Sun, Jie Na, Yue Huang, Guang Liu
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-06-01
Series:Frontiers in Cell and Developmental Biology
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Online Access:https://www.frontiersin.org/articles/10.3389/fcell.2025.1609826/full
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author Shaojun Wang
Yi Sun
Yi Sun
Jie Na
Yue Huang
Yue Huang
Guang Liu
Guang Liu
author_facet Shaojun Wang
Yi Sun
Yi Sun
Jie Na
Yue Huang
Yue Huang
Guang Liu
Guang Liu
author_sort Shaojun Wang
collection DOAJ
description Human embryonic stem cells (hESC)-derived retinal organoids are sophisticated in vitro systems for dissecting the complex dynamics of human retinal development. The formation of the human retina is a precisely organized process that depends on the regulated differentiation of retinal progenitor cells; however, many of the basic mechanisms remain to be explored. Here, using hESC-derived retinal organoids, we elucidated the temporal contribution of RAX2 to retinal development, with an emphasis on photoreceptor cells (PC) formation. The results were corroborated using human fetal retinal tissue at various gestational ages. Using CRISPR/Cas9-mediated gene knockout, we delineated the essential role of RAX2 in modulating PC specifications. RAX2 deficiency significantly altered the expression of PAX6 and SOX2, two essential regulators of retinogenesis. Our results suggested that RAX2 is significant in retinal development, underpinning its potential as a therapeutic target in related retinal disorders.
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publishDate 2025-06-01
publisher Frontiers Media S.A.
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series Frontiers in Cell and Developmental Biology
spelling doaj-art-15d70f5dfa2e4fdc9d4ee7bd4fb1fbbf2025-08-20T03:24:52ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2025-06-011310.3389/fcell.2025.16098261609826Molecular analysis of RAX2-regulated retinal development using human retinal organoids at a single-cell resolutionShaojun Wang0Yi Sun1Yi Sun2Jie Na3Yue Huang4Yue Huang5Guang Liu6Guang Liu7Senior Department of Ophthalmology, 3rd Medical Center of Chinese PLA General Hospital, Beijing, ChinaState Key Laboratory of Common Mechanism Research for Major Disease, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaDepartment of Medical Genetics, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaCenter for Regeneration, Aging and Chronic Diseases, School of Basic Medical Sciences, State Key Laboratory for Complex, Severe and Rare Diseases, Tsinghua University, Beijing, ChinaState Key Laboratory of Common Mechanism Research for Major Disease, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaDepartment of Medical Genetics, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaState Key Laboratory of Common Mechanism Research for Major Disease, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaDepartment of Medical Genetics, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaHuman embryonic stem cells (hESC)-derived retinal organoids are sophisticated in vitro systems for dissecting the complex dynamics of human retinal development. The formation of the human retina is a precisely organized process that depends on the regulated differentiation of retinal progenitor cells; however, many of the basic mechanisms remain to be explored. Here, using hESC-derived retinal organoids, we elucidated the temporal contribution of RAX2 to retinal development, with an emphasis on photoreceptor cells (PC) formation. The results were corroborated using human fetal retinal tissue at various gestational ages. Using CRISPR/Cas9-mediated gene knockout, we delineated the essential role of RAX2 in modulating PC specifications. RAX2 deficiency significantly altered the expression of PAX6 and SOX2, two essential regulators of retinogenesis. Our results suggested that RAX2 is significant in retinal development, underpinning its potential as a therapeutic target in related retinal disorders.https://www.frontiersin.org/articles/10.3389/fcell.2025.1609826/fullhuman embryonic stem cells (hESC)retinal organoidretinal developmentphotoreceptor cellsScRNA-seq
spellingShingle Shaojun Wang
Yi Sun
Yi Sun
Jie Na
Yue Huang
Yue Huang
Guang Liu
Guang Liu
Molecular analysis of RAX2-regulated retinal development using human retinal organoids at a single-cell resolution
Frontiers in Cell and Developmental Biology
human embryonic stem cells (hESC)
retinal organoid
retinal development
photoreceptor cells
ScRNA-seq
title Molecular analysis of RAX2-regulated retinal development using human retinal organoids at a single-cell resolution
title_full Molecular analysis of RAX2-regulated retinal development using human retinal organoids at a single-cell resolution
title_fullStr Molecular analysis of RAX2-regulated retinal development using human retinal organoids at a single-cell resolution
title_full_unstemmed Molecular analysis of RAX2-regulated retinal development using human retinal organoids at a single-cell resolution
title_short Molecular analysis of RAX2-regulated retinal development using human retinal organoids at a single-cell resolution
title_sort molecular analysis of rax2 regulated retinal development using human retinal organoids at a single cell resolution
topic human embryonic stem cells (hESC)
retinal organoid
retinal development
photoreceptor cells
ScRNA-seq
url https://www.frontiersin.org/articles/10.3389/fcell.2025.1609826/full
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