Analysis of macular ganglion cell status in dry AMD using single-nucleus transcriptomics

Age-related macular degeneration (AMD) is classically defined by photoreceptor and retinal pigment epithelium degeneration; however, increasing evidence suggests that retinal ganglion cells (RGCs) may also be affected. The RGCs, including intrinsically photosensitive RGCs (ipRGCs), play essential roles in visual and non-image-forming functions, yet their vulnerability in the human dry AMD macula remains poorly understood. Here, we performed exploratory single-nucleus RNA sequencing (snRNA-seq) of macular tissue from non-AMD (N = 2) and dry AMD (N = 2) human donor eyes to assess RGC populations and subtype-specific alterations.

Clustering and re-embedding identified distinct RGC transcriptional subtypes without a consistent pattern of overall RGC depletion in dry AMD maculae. Donor-level pseudobulk analyses showed broadly similar expression of pan-RGC markers and subtype-associated markers across non-AMD and dry AMD samples, with inter-donor variability. The ipRGCs, defined by OPN4 and EOMES expression, were detected in both groups, and SoupX-corrected transcript counts did not reveal a consistent change in ipRGCs in dry AMD donors.

Together these findings suggest that no gross depletion of RGCs, including melanopsin rich ipRGCs in the maculae of dry AMD donors in our current study cohort. Overall, the current pilot study provides preliminary insights into inner retinal neuronal characteristics in dry AMD and establishes a foundation for larger-cohort studies to determine whether ipRGCs are retained in advanced stages of AMD and whether residual ipRGC-mediated signaling may contribute to rudimentary visual awareness following extensive photoreceptor loss. The authors acknowledge Indiana University-sponsored Cohen pilot grant awarded to the corresponding author Tasneem P.

Sharma, for supporting this work. The authors also acknowledge the Indiana University School of Medicine Department of Ophthalmology Startup Funds and Research to Prevent Blindness Unrestricted funds to the department for supporting this work. The authors thank the donors and their families for generously providing tissues for research.

The authors thank Lions Gift of Sight, Minnesota for providing the donor eyes for performing this work. The authors acknowledge Medical Genomic Core facility at Indiana University for performing snRNA-seq. The authors acknowledge Dr.

Weiming Mao for providing their microscope facility. This work was supported by Indiana University-sponsored Cohen pilot grant awarded to the corresponding author Tasneem P. The work was also supported by the Indiana University School of Medicine Department of Ophthalmology Startup Funds and Research to Prevent Blindness Unrestricted funds.