Comprehensive analysis of m⁶A regulators characterized by the immune microenvironment in Duchenne muscular dystrophy

Duchenne muscular dystrophy (DMD) is an X-linked, incurable, degenerative neuromuscular disease that is exacerbated by secondary inflammation. N⁶-methyladenosine (m⁶A), the most common base modification of RNA, has pleiotropic immunomodulatory effects in many diseases. However, the role of m⁶A modification in the immune microenvironment of DMD remains elusive.

Our study retrospectively analyzed the expression data of 56 muscle tissues from DMD patients and 26 from non-muscular dystrophy individuals. Based on single sample gene set enrichment analysis, immune cells infiltration was identified and the result was validated by flow cytometry analysis and immunohistochemical staining. Then, we described the features of genetic variation in 26 m⁶A regulators and explored their relationship with the immune mircoenvironment of DMD patients through a series of bioinformatical analysis. At last, we determined subtypes of DMD patients by unsupervised clustering analysis and characterized the molecular and immune characteristics in different subgroups.

DMD patients have a sophisticated immune microenvironment that is significantly different from non-DMD controls. Numerous m⁶A regulators were aberrantly expressed in the muscle tissues of DMD and inversely related to most muscle-infiltrating immune cell types and immune response-related signaling pathways. A diagnostic model involving seven m⁶A regulators was established using LASSO. Furthermore, we determined three m⁶A modification patterns (cluster A/B/C) with distinct immune microenvironmental characteristics.

In summary, our study demonstrated that m⁶A regulators are intimately linked to the immune microenvironment of muscle tissues in DMD. These findings may facilitate a better understanding of the immunomodulatory mechanisms in DMD and provide novel strategies for the treatment.