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Gene expression profile at single cell level of mononuclear cells from skeletal muscle (gastrocnemius) of non-PAD controls and PAD patients

ABSTRACT: The molecular and cellular mechanisms of skeletal muscle from peripheral artery disease (PAD) patients are not completely understood. We used single cell RNA sequencing (scRNA-seq) to characterize the skeletal muscle diversity between control and PAD patients.

ORGANISM(S): Homo sapiens

PROVIDER: GSE235143 | GEO | 2023/10/05

REPOSITORIES: GEO

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Gene expression profile at single cell level of mononuclear cells from skeletal muscle (gastrocnemius) of non-PAD controls and PAD patients

Project description:Gene expression profile at single cell level of mononuclear cells from skeletal muscle (gastrocnemius) of non-PAD controls and PAD patients

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Single Nuclei RNA Sequencing of the Gastrocnemius Muscle Reveals Unique Transcriptomic Signatures of Peripheral Artery Disease

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Project description:Peripheral Artery Disease (PAD) is a disease characterized by atherosclerosis of the arteries supplying the lower extremities. In PAD, atherosclerotic occlusions restrict the delivery of blood to the skeletal muscles of affected extremities. As a result, the muscles which are supplied by the affected arteries can experience ischemia/reperfusion (I/R) injuries. This can lead to an altered metabolomics profile, increased oxidative damage to those muscles, and the degeneration of myofibers. In our study, we modeled I/R injuries by culturing differentiated skeletal muscle derived cells (skMDCs), also known as myotubes, under cyclic conditions of hypoxia and hyperoxia. These conditions mimic those that PAD patient muscles experience. Our results show an altered transcriptomic profile in PAD patient derived myotubes and explore the contribution of I/R injuries to those transcriptomic changes.

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Project description:Peripheral arterial disease (PAD), caused by atherosclerosis, leads to limb ischemia, muscle damage, and impaired mobility in the lower extremities. Recent studies suggest that circadian rhythm disruptions can hinder vascular repair during ischemia, but the specific tissues involved and the impact on muscle health remain unclear. This study investigates the role of the skeletal muscle circadian clock in muscle adaptation to ischemic stress using a surgical mouse model of hindlimb ischemia. We used mice with specific genetic loss of the circadian clock activator, BMAL1, in adult skeletal muscle tissues (Bmal1muscle). Bmal1muscle mice and controls underwent femoral artery ligation surgery to induce hindlimb ischemia. Laser doppler imaging was used to assess limb perfusion at various time points after the surgery. Muscle tissues were analyzed with RNA sequencing and histological examination to investigate PAD-related muscle pathologies. Additionally, we studied the role of BMAL1 in muscle fiber adaptation to hypoxia using RNA and ATAC sequencing analyses in primary myotube culture model. Disrupted expression of circadian rhythm-related genes was observed in existing RNA-seq datasets from PAD patient-derived endothelial cells and ischemic limb skeletal muscles. Genetic loss of Bmal1 specifically in adult mouse skeletal muscle tissues delayed reperfusion recovery following induction of hindlimb ischemia. Histological examination of muscle tissues showed reduced regenerated myofiber number and a decreased proportion of type IIB fast-twitch myofibers in Bmal1musc mouse muscles in the ischemic limbs, but not in their contralateral non-ischemic limbs. Transcriptomic analysis revealed abrogated metabolic, angiogenic, and myogenic pathways relevant to hypoxia-adaptation in Bmal1musc mouse muscles. These changes were corroborated in Bmal1-deficient cultured primary myotubes cultured under hypoxic conditions.

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Circadian Dysfunction in Skeletal Muscle Impairs Limb Perfusion and Muscle Regeneration in Peripheral Artery Disease [Myotube-ATACseq]

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Circadian Dysfunction in Skeletal Muscle Impairs Limb Perfusion and Muscle Regeneration in Peripheral Artery Disease [Myotube-RNAseq]

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