Disease modeling and gene correction of LGMDR21 iPSCs elucidates the role of POGLUT1 in skeletal muscle maintenance, regeneration, and the satellite cell niche.
Disease modeling and gene correction of LGMDR21 iPSCs elucidates the role of POGLUT1 in skeletal muscle maintenance, regeneration, and the satellite cell niche.
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DOI：
10.1016/j.omtn.2023.07.037
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发表时间：
2023-09-12
期刊：
影响因子：
--
通讯作者：
Darabi, Radbod
中科院分区：
文献类型：
--
作者：
Ortiz-Vitali, Jose L.;Wu, Jianbo;Xu, Nasa;Shieh, Annie W.;Niknejad, Nima;Takeuchi, Megumi;Paradas, Carmen;Lin, Chunru;Jafar-Nejad, Hamed;Haltiwanger, Robert S.;Wang, Sidney H.;Darabi, Radbod
常染色体隐性肢带型肌营养不良症 21 (LGMDR21) 是由蛋白质 O-葡萄糖基转移酶 1 (POGLUT1) 的致病性变异引起的，POGLUT1 负责在约 50 种哺乳动物蛋白质（包括 Notch 受体）中发现的特定表皮生长因子 (EGF) 重复序列的 O-葡萄糖基化。先前来自患者活检的数据表明，Notch 信号传导受损、肌肉干细胞减少和加速分化可能与疾病病因有关。使用患者诱导多能干细胞 (iPSC)、其校正的同种型和对照 iPSC，基因表达谱表明 POGLUT1、NOTCH、肌肉发育、细胞外基质 (ECM)、细胞粘附和迁移的失调作为相关途径。它们还表现出体外 POGLUT1 酶活性和 NOTCH 信号传导降低以及肌生成、增殖、迁移和分化缺陷。此外，体内研究表明，植入、肌肉干细胞形成、PAX7 表达和维持显着减少，同时间质中错误定位的 PAX7+ 细胞百分比增加。使用 CRISPR-Cas9 切口酶对患者 iPSC 进行基因校正，挽救了主要的体外和体内表型。这些结果证明了 iPSC 和基因校正在疾病建模和表型拯救中的功效，并提供了肌肉干细胞生态位定位、PAX7 表达和细胞迁移作为 LGMDR21 中可能机制的证据。 Darabi 及其同事使用 iPSC 研究一种新型肢带型肌营养不良症 (LGMDR21)，以便使用 CRIPSR-Cas9n 进行疾病建模和基因校正。这项研究强调了 iPSC 识别疾病机制的适用性以及基因校正对于逆转受影响的致病机制的功效。
Autosomal recessive limb-girdle muscular dystrophy 21 (LGMDR21) is caused by pathogenic variants in protein O-glucosyltransferase 1 (POGLUT1), which is responsible for O-glucosylation of specific epidermal growth factor (EGF) repeats found in ∼50 mammalian proteins, including Notch receptors. Previous data from patient biopsies indicated that impaired Notch signaling, reduction of muscle stem cells, and accelerated differentiation are probably involved in disease etiopathology. Using patient induced pluripotent stem cells (iPSCs), their corrected isotypes, and control iPSCs, gene expression profiling indicated dysregulation of POGLUT1, NOTCH, muscle development, extracellular matrix (ECM), cell adhesion, and migration as involved pathways. They also exhibited reduced in vitro POGLUT1 enzymatic activity and NOTCH signaling as well as defective myogenesis, proliferation, migration and differentiation. Furthermore, in vivo studies demonstrated significant reductions in engraftment, muscle stem cell formation, PAX7 expression, and maintenance, along with an increased percentage of mislocalized PAX7+ cells in the interstitial space. Gene correction in patient iPSCs using CRISPR-Cas9 nickase led to the rescue of the main in vitro and in vivo phenotypes. These results demonstrate the efficacy of iPSCs and gene correction in disease modeling and rescue of the phenotypes and provide evidence of the involvement of muscle stem cell niche localization, PAX7 expression, and cell migration as possible mechanisms in LGMDR21. Darabi and colleagues used iPSCs to study a new type of limb girdle muscular dystrophy (LGMDR21) for disease modeling and gene correction using CRIPSR-Cas9n. This study highlights the suitability of iPSCs to identify disease mechanisms and the efficacy of gene correction for reversing affected pathogenic mechanisms.
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