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Dominant RDH12-retinitis pigmentosa impairs photoreceptor development and cone function in retinal organoids

ABSTRACT: Retinal dehydrogenase 12 (RDH12) is a photoreceptor NADPH-dependent retinal reductase enzyme, which converts all-trans-retinal to all-trans-retinol. In rare cases, heterozygous variants in RDH12 are associated with a mild, late-onset autosomal dominant (AD) retinitis pigmentosa (RP). There are no dominant RDH12 animal models, hence, we generated retinal organoids from human induced pluripotent stem cells (hiPSC) derived from dermal fibroblasts from a RDH12-AD patient with the heterozygous pathogenic variant c.759del p.(Phe254Leufs*24) and an unrelated unaffected individual (WT). Retinal organoids were characterised at different stages of differentiation; at 18-weeks there were no differences in rod and cone photoreceptors with correct localisation of RDH12 in the photoreceptor inner segments in both WT and RDH12-AD. Transmission electron microscopy on week 37 retinal organoids showed a normal outer limiting membrane and mitochondria, but photoreceptors were 1.8 times less abundant and shorter in RDH12-AD mutant compared to WT. Transcriptomic analyses on week 44 retinal organoids revealed visual cone function, retinol biosynthesis and the vitamin A pathway are highly affected in RDH12-AD retinal organoids. Our study is the first to describe a human RDH12 retinal organoid model, and we found subtle differences between the WT and mutant at later stages of photoreceptor differentiation. This is in keeping with the milder disease course seen in humans, but it provides insights into the aetiology and possible targets for future therapeutic development.

ORGANISM(S): Homo sapiens

PROVIDER: GSE271751 | GEO | 2025/05/28

REPOSITORIES: GEO

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Project description:The loss of cone photoreceptor cells, which are critical for optimal daylight vision, have the great impact on vision during retinal degenerations. Retinal differentiation of human induced pluripotent stem cell (hiPSC) sources could provide a renewable source of cone photoreceptors towards developing a cone cell replacement therapy to treat blindness. Demonstration of comparable gene expression profiles between human foetal and stem cell-derived cones at equivalent stages is required to progress the cell transplantation approach into the patient, as it is hypothesised stem cell-derived cones are required to show high levels of developmental recapitulation of the in vivo generated cones. In this study, the AAV2/9.pR2.1.GFP reporter was used to specifically label L/M-opsin cone photoreceptors in human foetal retinal samples, obtained from the MRC-Wellcome Trust Human Developmental Biology Resource, at a range of developmental stages. The L/M-opsin cone population represent the majority cone cell types in the adult human retina and are the only photoreceptors present within the fovea. Using fluorescence activated cell sorting, L/M-opsin GFP+ cones and GFP- retinal populations, alongside total foetal retinal samples containing all retinal cell tytpes, were isolated and processed for bulk RNA sequencing and downstream comparative analysis. Using DESeq2 differential gene expression analyses, statistically significant genes enriched within the GFP+ human foetal LM-opsin cone populations were determined which led to the identification of a cone enriched gene signature of human L/M-opsin cone photoreceptors. The AAV2/9.pR2.1.GFP reporter was applied to hiPSC-derived retinal cultures to isolate and process cone-like cell populations for RNA sequencing using the same strategy developed within the human foetal retina. Applying the cone enriched gene signature to the transcriptome of hiPSC-derived GFP+ samples at equivalent developmental stages revealed some expression similarities in genes found to be enriched within the late foetal L/M-opsin cone photoreceptors. This analysis overall revealed an intermediate stage of cone differentiation was achieved within the hiPSC-derived samples and the comparison to human foetal L/M-opsin gene express profiles suggesting further differentiation of hiPSC-derived sample is required.

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Dominant RDH12-retinitis pigmentosa impairs photoreceptor development and cone function in retinal organoids

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