Hypoxia enhances human myoblast differentiation: involvement of HIF1α and impact of DUX4, the FSHD causal gene

IF 5.3 2区医学 Q2 CELL BIOLOGY

Skeletal Muscle Pub Date : 2023-12-16 DOI:10.1186/s13395-023-00330-2

Thuy-Hang Nguyen, Lise Paprzycki, Alexandre Legrand, Anne-Emilie Declèves, Philipp Heher, Maelle Limpens, Alexandra Belayew, Christopher R. S. Banerji, Peter S. Zammit, Alexandra Tassin

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引用次数: 0

Abstract

Hypoxia is known to modify skeletal muscle biological functions and muscle regeneration. However, the mechanisms underlying the effects of hypoxia on human myoblast differentiation remain unclear. The hypoxic response pathway is of particular interest in patients with hereditary muscular dystrophies since many present respiratory impairment and muscle regeneration defects. For example, an altered hypoxia response characterizes the muscles of patients with facioscapulohumeral dystrophy (FSHD). We examined the impact of hypoxia on the differentiation of human immortalized myoblasts (LHCN-M2) cultured in normoxia (PO2: 21%) or hypoxia (PO2: 1%). Cells were grown in proliferation (myoblasts) or differentiation medium for 2 (myocytes) or 4 days (myotubes). We evaluated proliferation rate by EdU incorporation, used myogenin-positive nuclei as a differentiation marker for myocytes, and determined the fusion index and myosin heavy chain-positive area in myotubes. The contribution of HIF1α was studied by gain (CoCl2) and loss (siRNAs) of function experiments. We further examined hypoxia in LHCN-M2-iDUX4 myoblasts with inducible expression of DUX4, the transcription factor underlying FSHD pathology. We found that the hypoxic response did not impact myoblast proliferation but activated precocious myogenic differentiation and that HIF1α was critical for this process. Hypoxia also enhanced the late differentiation of human myocytes, but in an HIF1α-independent manner. Interestingly, the impact of hypoxia on muscle cell proliferation was influenced by dexamethasone. In the FSHD pathological context, DUX4 suppressed HIF1α-mediated precocious muscle differentiation. Hypoxia stimulates myogenic differentiation in healthy myoblasts, with HIF1α-dependent early steps. In FSHD, DUX4-HIF1α interplay indicates a novel mechanism by which DUX4 could interfere with HIF1α function in the myogenic program and therefore with FSHD muscle performance and regeneration.

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缺氧可促进人类肌母细胞分化：HIF1α的参与和FSHD致病基因DUX4的影响

众所周知，缺氧会改变骨骼肌的生物功能和肌肉再生。然而，低氧对人类成肌细胞分化的影响机制仍不清楚。遗传性肌肉萎缩症患者对低氧反应途径尤为关注，因为许多患者存在呼吸障碍和肌肉再生缺陷。例如，面岬肱肌营养不良症（FSHD）患者的肌肉就具有低氧反应改变的特征。我们研究了低氧对在常氧（PO2：21%）或低氧（PO2：1%）条件下培养的人类永生肌母细胞（LHCN-M2）分化的影响。细胞在增殖培养基（肌母细胞）或分化培养基中培养 2 天（肌细胞）或 4 天（肌管）。我们用 EdU 结合评估增殖率，用肌原蛋白阳性核作为肌细胞的分化标记，并测定肌管的融合指数和肌球蛋白重链阳性面积。通过功能增益（CoCl2）和功能缺失（siRNAs）实验研究了HIF1α的贡献。我们进一步研究了LHCN-M2-iDUX4肌母细胞的缺氧反应，这些肌母细胞诱导性表达了DUX4，DUX4是FSHD病理学的基础转录因子。我们发现，缺氧反应不会影响肌母细胞的增殖，但会激活早熟的肌原分化，而HIF1α对这一过程至关重要。缺氧也增强了人类肌细胞的后期分化，但其方式与 HIF1α 无关。有趣的是，低氧对肌肉细胞增殖的影响受到地塞米松的影响。在前列腺增生症的病理环境中，DUX4抑制了HIF1α介导的肌肉早熟分化。缺氧会刺激健康肌母细胞的成肌分化，其早期步骤依赖于 HIF1α。在前列腺增生症中，DUX4-HIF1α的相互作用表明了一种新的机制，通过这种机制，DUX4可以干扰肌生成程序中HIF1α的功能，从而影响前列腺增生症肌肉的性能和再生。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Skeletal Muscle CELL BIOLOGY-

CiteScore

9.10

自引率

0.00%

发文量

审稿时长

12 weeks

期刊介绍： The only open access journal in its field, Skeletal Muscle publishes novel, cutting-edge research and technological advancements that investigate the molecular mechanisms underlying the biology of skeletal muscle. Reflecting the breadth of research in this area, the journal welcomes manuscripts about the development, metabolism, the regulation of mass and function, aging, degeneration, dystrophy and regeneration of skeletal muscle, with an emphasis on understanding adult skeletal muscle, its maintenance, and its interactions with non-muscle cell types and regulatory modulators. Main areas of interest include: -differentiation of skeletal muscle- atrophy and hypertrophy of skeletal muscle- aging of skeletal muscle- regeneration and degeneration of skeletal muscle- biology of satellite and satellite-like cells- dystrophic degeneration of skeletal muscle- energy and glucose homeostasis in skeletal muscle- non-dystrophic genetic diseases of skeletal muscle, such as Spinal Muscular Atrophy and myopathies- maintenance of neuromuscular junctions- roles of ryanodine receptors and calcium signaling in skeletal muscle- roles of nuclear receptors in skeletal muscle- roles of GPCRs and GPCR signaling in skeletal muscle- other relevant aspects of skeletal muscle biology. In addition, articles on translational clinical studies that address molecular and cellular mechanisms of skeletal muscle will be published. Case reports are also encouraged for submission. Skeletal Muscle reflects the breadth of research on skeletal muscle and bridges gaps between diverse areas of science for example cardiac cell biology and neurobiology, which share common features with respect to cell differentiation, excitatory membranes, cell-cell communication, and maintenance. Suitable articles are model and mechanism-driven, and apply statistical principles where appropriate; purely descriptive studies are of lesser interest.