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Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Animal Models

ABSTRACT: PROM1 (CD133/AC133) is widely expressed across various tissues, yet paradoxically, its mutations are exclusively associated with retinal degeneration. The mechanisms underlying this retina-specific vulnerability remain poorly understood, and no effective treatment currently exists for PROM1-associated inherited retinal dystrophies (IRDs). To address these gaps, we employed retinal organoids (ROs) to delineate the spatiotemporal expression patterns of PROM1, identifying its co-localization with the ciliary marker ARL13B and the outer segment (OS) marker PRPH2 during photoreceptor differentiation. Furthermore, using hiPSCs, hiPSC-RPE cells, and ROs, we discovered photoreceptor-specific mRNA alternative splicing isoforms and distinct protein N-glycosylation patterns of PROM1. To investigate disease mechanisms and therapeutic potential, personalized hiPSCs and ROs were derived from an individual with a PROM1 c.619G>T (p.E207X) homozygous mutation and his unaffected heterozygous mother. The mutation induced pronounced nonsense-mediated decay (NMD) and nonsense-associated altered splicing (NAS), resulting in complete loss of PROM1 protein and severe disruption of OS-like structure in patient-derived ROs (Hom-ROs). To rescue these defects, a photoreceptor-specific AAV7m8-CRXp-hPROM1 vector was engineered, which successfully restored PROM1 expression and OS-like structure in Hom-ROs. Therapeutic efficacy was further validated in vivo using Prom1-/- mice, where subretinal delivery of AAV8-CRXp-hPROM1 achieved robust, photoreceptor-specific expression of human PROM1, significantly preserving OS structure and visual function. Our study establishes a mechanistic link between PROM1 deficiency and photoreceptor pathology and provide the first preclinical evidence supporting gene augmentation therapy for PROM1-IRD, highlighting its translational potential.

ORGANISM(S): Homo sapiens

PROVIDER: GSE299857 | GEO | 2026/06/18

REPOSITORIES: GEO

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Exogenous Photoreceptor-Specific N-Glycosylated PROM1 Rescues Retinal Degeneration in Patient and Animal Models

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