Comprehensive Analysis of N6-Methyladenosine Modification Profiling in Diabetic Erectile Dysfunction.

Abstract

Purpose: Diabetic erectile dysfunction (DMED) is a prevalent condition with limited treatment options. The role of RNA N6-methyladenosine (m⁶A) modification in the pathogenesis of DMED remains elusive. This study aimed to investigate the underlying m⁶A modification patterns and identify potential therapeutic targets for DMED.

Materials and methods: A rat model of DMED was established using streptozotocin injection and confirmed by apomorphine-induced penile erection. Erectile function was assessed via cavernous nerve electrostimulation. Fibrosis in the corpus cavernosum was evaluated using Masson's trichrome staining. RNA m⁶A modification levels and the expression of associated methyltransferases were examined by dot blot and quantitative real-time PCR. MeRIP-seq and RNA-seq were employed to identify differentially methylated and expressed genes. Conjoint analysis was performed to explore associated biological processes and identify key genes, which were subsequently validated.

Results: Elevated levels of RNA m⁶A modification were observed in DMED, accompanied by altered expression of METTL14 and METTL3. A total of 2,789 genes associated with 3574 m⁶A peaks were identified (p<0.05). Differentially methylated m⁶A genes were implicated in muscle cell differentiation, cell junction organization, and Wnt signaling pathways. Combined analysis of MeRIP-seq and RNA-seq identified and validated POSTN and LOX as key genes. These genes were associated with fibrosis, cell-matrix adhesion, and regulated Notch signaling pathway, and were predominantly enriched in corpus cavernosum fibroblasts of DMED.

Conclusions: This exploratory study provides the first exploration of RNA m⁶A modification in DMED, and offers novel insights into the pathogenesis of DMED and potential therapeutic targets.