Identification of biomarkers associated with pathological dynamic progression in dystrophinopathies

Dystrophinopathies, including Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD), are X-linked disorders characterized by progressive muscle degeneration. However, reliable molecular markers that reflect pathological progression and disease severity remain limited. In this study, 33 muscle samples from the Gene Expression Omnibus were analyzed, including control subjects (n = 6) and dystrophinopathy cases (n = 27) stratified by pathological severity. Differential expression and trend analyses identified 64 severity-associated genes (58 upregulated and 6 downregulated). Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis indicated significant involvement of cytoskeleton-related pathways. Nine core genes (COL1A1, COL1A2, COL3A1, COL5A1, COL5A2, COL6A6, MYH3, MYH8, and VCAN) exhibited progressive upregulation with increasing disease severity. Immune infiltration analysis revealed enrichment of M2 macrophages, resting dendritic cells, and resting mast cells, along with depletion of plasma cells, regulatory T cells, and neutrophils. Validation using five independent datasets and mdx mouse muscle samples supported the dysregulation of these core genes at the transcriptomic level, with partial concordance observed between human and mouse dystrophinopathy models. Furthermore, integration of gene expression and immune infiltration features improved the accuracy of dystrophinopathy severity assessment. These findings suggest that extracellular matrix-related genes may serve as potential biomarkers for monitoring disease progression across dystrophinopathies and provide insights into disease-associated molecular alterations.