5-Aza-2’-Deoxycytidine Treatment Induces Müller Glia of Rhodopsin Deficient Mice Transdifferentiation Into Photoreceptor Lineage Cells

Abstract

Purpose : Purpose: Müller glia in lower vertebrates exhibit the ability to repair and regenerate retinal cells by turning into progenitor cells and generating new photoreceptors post-injury. In contrast, mammalian Müller glia display limited regenerative capacity, partly due to constrained epigenetic plasticity. This study aimed to explore the potential of Decitabine, a DNA methyltransferase (DNMT) inhibitor, in awakening the transdifferentiation potential of Müller glia, especially in mice with inherited photoreceptor degeneration.

Methods : Methods: Primary Müller glia isolated from postnatal day 12 (P12) rhodopsin knockout mice were cultured and treated with 5-Aza-2’-Deoxycytidine (5-AZA-CdR-Decitabine; 10 μM). The expression of Müller glia, retinal progenitor cell, and photoreceptor cell markers were evaluated 6 days post Decitabine treatment using qPCR and immunocytochemistry to assess Müller glial transdifferentiation. Statistical analysis was conducted using a two-tailed Student’s t-test or ANOVA.

Results : Results: Decitabine treatment to cultured Müller glia significantly upregulated CRX (photoreceptor-specific transcription factor) and Recoverin (photoreceptor-specific cell marker) alongside a modest upregulation of Sox2 (retinal Müller glial and progenitor cell marker), and CHX10 (retinal progenitor marker), suggesting Müller glia dedifferentiation and transdifferentiation into cells of photoreceptor cell lineage. Immunocytochemistry data further confirmed the increased Recoverin and Sox2 expression in Decitabine treated primary Müller glia cultures.

Conclusions : Conclusions: These data indicate the possible role of DNA demethylation in promoting Müller glia differentiation. Further elucidation of the epigenetic landscape and its targeted modification may uncover the molecular insight into the regulation of Müller glial responses to injury and advance therapeutic strategies for effective management of retinal diseases.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.