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Published on: September 18, 2019
Transcriptomic Profiling Identifies Ferroptosis-Related Gene Signatures in Ischemic Muscle Satellite Cells Affected
Lillian Tran1, Bowen Xie1, Edwyn Assaf1
1Department of Surgery, University of Pittsburgh Medical Center, PA (L.T., B.X., E.A., R.F., R.I.M.A., U.S.).
Background:
We hypothesized that transcriptomic profiling of muscle satellite cells in peripheral artery disease (PAD) would identify damage-related pathways contributing to skeletal muscle myopathy. We identified a potential role for ferroptosis-a form of programmed lytic cell death by iron-mediated lipid peroxidation-as one such pathway. Ferroptosis promotes myopathy in ischemic cardiac muscle but has an unknown role in PAD.
Methods:
Muscle satellite cells from donors with PAD were obtained during surgery. cDNA libraries were processed for single-cell RNA sequencing using the 10X Genomics platform. Protein expression was confirmed based on pathways inferred by transcriptomic analysis.
Results:
Unsupervised cluster analysis of over 25 000 cells aggregated from 8 donor samples yielded distinct cell populations grouped by a shared unique transcriptional fingerprint. Quiescent cells were diminished in ischemic muscle while myofibroblasts and apoptotic cells were prominent. Differential gene expression demonstrated a surprising increase in genes associated with iron transport and oxidative stress and a decrease in GPX4 (glutathione peroxidase 4) in ischemic PAD-derived cells. Release of the danger signal HMGB1 (high mobility group box-1) correlated with ferroptotic markers including surface transferrin receptor and were higher in ischemia. Furthermore, lipid peroxidation in muscle satellite cells was modulated by ferrostatin, a ferroptosis inhibitor. Histology confirmed iron deposition and lipofuscin, an inducer of ferroptosis in PAD-affected muscle.
Conclusions:
This report presents a novel finding that genes known to be involved in ferroptosis are differentially expressed in human skeletal muscle affected by PAD. Targeting ferroptosis may be a novel therapeutic strategy to reduce PAD myopathy.
Insights
Peripheral artery disease (PAD) muscle satellite cells show increased ferroptosis markers, suggesting this programmed cell death pathway contributes to skeletal muscle myopathy. Targeting ferroptosis may offer a new therapeutic approach for PAD.
Area of Science:
- Cellular Biology
- Molecular Medicine
- Pathophysiology
Background:
- Peripheral artery disease (PAD) is associated with skeletal muscle myopathy.
- The specific molecular pathways contributing to PAD-induced myopathy are not fully understood.
- Ferroptosis, a form of programmed cell death involving iron-mediated lipid peroxidation, is implicated in cardiac myopathy but its role in PAD is unknown.
Purpose of the Study:
- To investigate transcriptomic profiles of muscle satellite cells in PAD patients.
- To identify damage-related pathways contributing to skeletal muscle myopathy in PAD.
- To explore the potential role of ferroptosis in PAD-associated myopathy.
Main Methods:
- Single-cell RNA sequencing of muscle satellite cells from PAD donors.
- Analysis of gene expression patterns to identify cell populations and pathways.
- Confirmation of protein expression and pathway inference.
- Histological analysis for iron deposition and lipofuscin.
Main Results:
- Distinct cell populations were identified, with diminished quiescent cells and increased myofibroblasts and apoptotic cells in ischemic muscle.
- Genes associated with iron transport and oxidative stress were upregulated, while GPX4 expression decreased in PAD-derived cells.
- Ferroptotic markers, including HMGB1 release and transferrin receptor expression, were elevated in ischemic muscle.
- Iron deposition and lipofuscin were confirmed in PAD-affected muscle tissue.
Conclusions:
- Genes involved in ferroptosis are differentially expressed in human skeletal muscle affected by PAD.
- Ferroptosis plays a significant role in the skeletal muscle myopathy observed in PAD.
- Targeting ferroptosis presents a novel therapeutic strategy for mitigating PAD-related myopathy.
