Uncovering the Gene Regulatory Network of Endothelial Cells in Mouse Duchenne Muscular Dystrophy: Insights from Single-Nuclei RNA Sequencing Analysis

Abstract

Introduction: Duchenne muscular dystrophy (DMD) is a severe X-linked recessive disorder caused by mutations in the dystrophin gene, which leads to heart and respiratory failure. Despite the critical impact of DMD on endothelial cells (ECs), there is limited understanding of its effect on the endothelial gene network. The aim of this study was to investigate the impact of DMD on the gene regulatory network of ECs. Methods and Results: To gain insights into the role of the dystrophin muscular dystrophy gene (DMD) in ECs from Duchenne muscular dystrophy; the study utilized single-nuclei RNA sequencing (snRNA-seq) to evaluate the transcriptomic profile of ECs from skeletal muscles in DMD mutant mice (DMDmut) and wild-type control mice. The analysis showed that the DMD mutation resulted in the suppression of several genes, including SPTBN1 and the upregulation of multiple long noncoding RNAs (lncRNAs). GM48099, GM19951, and GM15564 were consistently upregulated in ECs and skeletal muscle cells from DMDmut, indicating that these dysregulated lncRNAs are conserved across different cell types. Gene ontology (GO) enrichment analysis revealed that the DMD mutation activated the following four pathways in ECs: fibrillary collagen trimer, banded collagen fibril, complex of collagen trimers, and purine nucleotide metabolism. The study also found that the metabolic pathway activity of ECs was altered. Oxidative phosphorylation (OXPHOS), fatty acid degradation, glycolysis, and pyruvate metabolism were decreased while purine metabolism, pyrimidine metabolism, and one carbon pool by folate were increased. Moreover, the study investigated the impact of the DMD mutation on ECs from skeletal muscles and found a significant decrease in their overall number, but no change in their proliferation. Conclusions: Overall, this study provides new insights into the gene regulatory program in ECs in DMD and highlights the importance of further research in this area.

Cellular interactions and microenvironment dynamics in skeletal muscle regeneration and disease

“…mdx mice revealed that ECs are enriched in pathways related to fibrillar collagen production, while pathways related to signaling receptor activity and molecular transduction were suppressed ( Saleh et al, 2022 ). Conversely, crucial genes responsible for EC function were downregulated, indicating potential functional and signaling deficits together with increased propensity to fibrosis ( Shen et al, 2023 ). At the same time, single-cell RNA sequencing showed that the proportion of distinct capillary EC populations changes drastically between healthy and dystrophic muscles, presenting a reduced number in mdx and D2.…”

Editorial: Special Issue—Understanding and Targeting Heart Failure: From Biology to Therapeutics

“…Shen et al [1] employed single-nuclei RNA sequencing (snRNA-seq) to evaluate the transcriptomic profile of endothelial cells (ECs) from skeletal muscles in Duchenne muscular dystrophy (DMD) mutant mice. The study showed that unique phenotypes in ECs, including suppressed expression of SPTBN1 and the upregulated expression of multiple long noncoding RNAs (lncRNAs), altered metabolic activity.…”