scholarly journals Human retinal organoids release extracellular vesicles that regulate gene expression in target human retinal progenitor cells

2021 ◽  
Author(s):  
Jing Zhou ◽  
Miguel Flores-Bellver ◽  
Jianbo Pan ◽  
Alberto Benito-Martin ◽  
Cui Shi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Progenitor Cells ◽  
Extracellular Vesicles ◽  
Noncoding Rna ◽  
Tissue Characterization ◽  
Retinal Development ◽  
Retinal Progenitor Cells ◽  
Retina Development ◽  
Retinal Tissue ◽  
Retinal Progenitor

AbstractThe mechanisms underlying retinal development have not been completely elucidated. Extracellular vesicles (EVs) are novel essential mediators of cell-to-cell communication with emerging roles in developmental processes. Nevertheless, the identification of EVs in human retinal tissue, characterization of their cargo, and analysis of their potential role in retina development has not been accomplished. Three-dimensional retinal tissue derived from human induced pluripotent stem cells (hiPSC) provide an ideal developmental system to achieve this goal. Here we report that hiPSC-derived retinal organoids release exosomes and microvesicles with small noncoding RNA cargo. EV miRNA cargo-predicted targetome correlates with GO pathways involved in mechanisms of retinogenesis relevant to specific developmental stages corresponding to hallmarks of native human retina development. Furthermore, uptake of EVs by human retinal progenitor cells leads to changes in gene expression correlated with EV miRNA cargo predicted gene targets, and mechanisms involved in retinal development, ganglion cell and photoreceptor differentiation and function.

scholarly journals Human Retinal Organoids Release Extracellular Vesicles That Regulate Gene Expression in Target Human Retinal Progenitor Cells

2021 ◽  
Author(s):  
Jing Zhou ◽  
Miguel Flores-Bellver ◽  
Jianbo Pan ◽  
Alberto Benito-Martin ◽  
Cui Shi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Progenitor Cells ◽  
Extracellular Vesicles ◽  
Noncoding Rna ◽  
Tissue Characterization ◽  
Retinal Development ◽  
Retinal Progenitor Cells ◽  
Retina Development ◽  
Retinal Tissue ◽  
Retinal Progenitor

Abstract The mechanisms underlying retinal development have not been completely elucidated. Extracellular vesicles (EVs) are novel essential mediators of cell-to-cell communication with emerging roles in developmental processes. Nevertheless, the identification of EVs in human retinal tissue, characterization of their cargo, and analysis of their potential role in retina development has not been accomplished. Three-dimensional retinal tissue derived from human induced pluripotent stem cells (hiPSC) provide an ideal developmental system to achieve this goal. Here we report that hiPSC-derived retinal organoids release exosomes and microvesicles with small noncoding RNA cargo. EV miRNA cargo-predicted targetome correlates with GO pathways involved in mechanisms of retinogenesis relevant to specific developmental stages corresponding to hallmarks of native human retina development. Furthermore, uptake of EVs by human retinal progenitor cells leads to changes in gene expression correlated with EV miRNA cargo predicted gene targets, and mechanisms involved in retinal development, ganglion cell and photoreceptor differentiation and function.

Gene expression is dynamically regulated in retinal progenitor cells prior to and during overt cellular differentiation

2014 ◽  
Vol 14 (1) ◽  
pp. 42-54 ◽  
Author(s):  
Rajiv Dixit ◽  
Nobuhiko Tachibana ◽  
Yacine Touahri ◽  
Dawn Zinyk ◽  
Cairine Logan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Progenitor Cells ◽  
Cellular Differentiation ◽  
Retinal Progenitor Cells ◽  
Retinal Progenitor

p57(Kip2) regulates progenitor cell proliferation and amacrine interneuron development in the mouse retina

Development ◽  
2000 ◽  
Vol 127 (16) ◽  
pp. 3593-3605 ◽  
Author(s):  
M.A. Dyer ◽  
C.L. Cepko
Keyword(s):  
Cell Cycle ◽  
Progenitor Cells ◽  
Kinase Inhibitor ◽  
Retinal Development ◽  
Cell Types ◽  
Cell Cycle Exit ◽  
Retinal Progenitor Cells ◽  
Mouse Development ◽  
Retinal Progenitor ◽  
Cyclin Kinase Inhibitor

A precise balance between proliferation and differentiation must be maintained during retinal development to obtain the correct proportion of each of the seven cell types found in the adult tissue. Cyclin kinase inhibitors can regulate cell cycle exit coincident with induction of differentiation programs during development. We have found that the p57(Kip2) cyclin kinase inhibitor is upregulated during G(1)/G(0) in a subset of retinal progenitor cells exiting the cell cycle between embryonic day 14.5 and 16.5 of mouse development. Retroviral mediated overexpression of p57(Kip2) in embryonic retinal progenitor cells led to premature cell cycle exit. Retinae from mice lacking p57(Kip2) exhibited inappropriate S-phase entry and apoptotic nuclei were found in the region where p57(Kip2) is normally expressed. Apoptosis precisely compensated for the inappropriate proliferation in the p57(Kip2)-deficient retinae to preserve the correct proportion of the major retinal cell types. Postnatally, p57(Kip2) was found to be expressed in a novel subpopulation of amacrine interneurons. At this stage, p57(Kip2)did not regulate proliferation. However, perhaps reflecting its role during this late stage of development, animals lacking p57(Kip2) showed an alteration in amacrine subpopulations. p57(Kip2) is the first gene to be implicated as a regulator of amacrine subtype/subpopulation development. Consequently, we propose that p57(Kip2) has two roles during retinal development, acting first as a cyclin kinase inhibitor in mitotic progenitor cells, and then playing a distinct role in neuronal differentiation.

scholarly journals Human embryonic stem cell-derived extracellular vesicles alleviate retinal degeneration by upregulating Oct4 to promote retinal Müller cell retrodifferentiation via HSP90

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yifeng Ke ◽  
Xiaoe Fan ◽  
Rui Hao ◽  
Lijie Dong ◽  
Min Xue ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Extracellular Vesicles ◽  
Fluorescence Intensity ◽  
Embryonic Stem ◽  
Müller Cells ◽  
Muller Cells ◽  
Retinal Progenitor Cells ◽  
Retinal Progenitor ◽  
Hsp90 Expression ◽  
Rcs Rats

Abstract Objective Retinal degenerative diseases remain the dominant causes of blindness worldwide, and cell replacement is viewed as a promising therapeutic direction. However, the resources of seed cells are hard to obtain. To further explore this therapeutic approach, human embryonic stem extracellular vesicles (hESEVs) were extracted from human embryonic stem cells (hESCs) to inspect its effect and the possible mechanism on retinal Müller cells and retinal function. Methods hESEVs were extracted by multi-step differential centrifugation, whose morphologies and specific biomarkers (TSG101, CD9, CD63, and CD81) were observed and measured. After hESEVs were injected into the vitreous cavity of RCS rats, the retinal tissues and retinal functions of rats were assessed. The alteration of Müller cells and retinal progenitor cells was also recorded. Microvesicles (MVs) or exosomes (EXOs) were extracted from hESCs transfected with sh-HSP90 or pcDNA3.1-HSP9, and then incubated with Müller cells to measure the uptake of EVs, MVs, or EXOs in Müller cells by immunofluorescence. The retrodifferentiation of Müller cells was determined by measuring Vimentin and CHX10. qRT-PCR and western blot were used to detect HSP90 expression in MVs and evaluate Oct4 level in Müller cells, and Co-IP to inspect the interaction of HSP90 and Oct4. Results RCS rats at the postnatal 30 days had increased retinal progenitor cells which were dedifferentiated from Müller cells. hESEVs were successfully extracted from hESCs, evidenced by morphology observation and positive expressions of specific biomarkers (TSG101, CD9, CD63, and CD81). hESEVs promoted Müller cells dedifferentiated and retrodifferentiated into retinal progenitor cells evidenced by the existence of a large amount of CHX10-positive cells in the retinal inner layer of RCS rats in response to hESEV injection. The promotive role of hESEVs was exerted by MVs demonstrated by elevated fluorescence intensity of CHX10 and suppressed Vimentin fluorescence intensity in MVs rather than in EXOs. HSP90 in MVs inhibited the retrodifferentiation of Müller cells and suppressed the expression level of Oct4 in Müller cells. Co-IP revealed that HSP90 can target Oct4 in Müller cells. Conclusion hESEVs could promote the retrodifferentiation of Müller cells into retinal progenitor cells by regulating the expression of Oct4 in Müller cells by HSP90 mediation in MVs.

scholarly journals Construction and Characterization of EGFP Reporter Plasmid Harboring Putative Human RAX Promoter for in vitro Monitoring of Retinal Progenitor Cells Identity

2020 ◽  
Author(s):  
Atefeh Atefi ◽  
Pendar Shojaei kojouri ◽  
Fereshteh Karamali ◽  
Shiva Irani ◽  
Mohammad Hossein Nasr Esfahani
Keyword(s):  
Progenitor Cells ◽  
Molecular Mechanisms ◽  
Embryonic Stem ◽  
Retinal Layer ◽  
Retinal Progenitor Cells ◽  
Regulatory Regions ◽  
Retina Development ◽  
Retinal Progenitor ◽  
Egfp Reporter

Abstract Background : In retinal degenerative disease, progressive and debilitating conditions result in deterioration of retinal cells and visual loss. In human, retina lacks the inherent capacity for regeneration. Therefore, regeneration of retinal layer from human retinal progenitor cells (hRPCs) is a challenging task and restricted in vitro maintenance of hRPCs remains as the main hurdle. Retina and anterior neural fold homeobox gene ( RAX ) play critical roles in developing retina and maintenance of hRPCs. In this study, for the first time regulatory regions of human RAX gene with potential promoter activity were experimentally investigated. Results: For this purpose, after in silico analysis of regulatory regions of human RAX gene, the expression of EGFP reporter derived by putative promoter sequences was first evaluated in 293T cells and then in hRPCS derived from human embryonic stem cells. The candidate region ( RAX -3258bp) showed the highest EGFP expression in hRPCs. This reporter construct can be used for invitro monitoring of hRPC identity and verification of an efficient culture medium for maintenance of these cells. Conclusions: Furthermore, our findings provide a platform for better insight into regulatory regions of human RAX gene and molecular mechanisms underlying its vital functions in retina development.

scholarly journals Individual Retinal Progenitor Cells Display Extensive Heterogeneity of Gene Expression

PLoS ONE ◽  
2008 ◽  
Vol 3 (2) ◽  
pp. e1588 ◽  
Author(s):  
Jeffrey M. Trimarchi ◽  
Michael B. Stadler ◽  
Constance L. Cepko
Keyword(s):  
Gene Expression ◽  
Progenitor Cells ◽  
Retinal Progenitor Cells ◽  
Retinal Progenitor
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