Functional analysis of iPSC-derived myocytes from a patient with carnitine palmitoyltransferase II deficiency
Functional analysis of iPSC-derived myocytes from a patient with carnitine palmitoyltransferase II deficiency
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DOI：
10.1016/j.bbrc.2014.04.084
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发表时间：
2014-05-30
影响因子：
3.1
通讯作者：
Kaneoka, Hidetoshi
中科院分区：
文献类型：
--
作者：
Yasuno, Tetsuhiko;Osafune, Kenji;Kaneoka, Hidetoshi
简介：肉碱棕榈酰转移酶 II (CPT II) 缺乏症是一种涉及长链脂肪酸 (FAO) β-氧化的遗传性疾病，可导致横纹肌溶解和随后的急性肾功能衰竭。疾病发病机制的详细机制仍不清楚；然而，用于研究的相关人类细胞类型（例如骨骼肌细胞）的可用性是有限的，需要开发新的疾病模型。 方法：我们从一名患有 CPT II 缺陷的日本患者的皮肤成纤维细胞中产生了人类诱导多能干细胞 (hiPSC)。通过引入肌原性分化 1 (MYOD1)（肌细胞分化的主要转录调节因子），将成熟肌细胞与患者来源的 hiPSC 进行分化。使用体外酰基肉碱分析测定，我们研究了降血脂药物苯扎贝特和热应激对 CPT II 缺陷型肌细胞和对照中线粒体FAO的影响。结果：CPT II 缺陷型肌细胞比对照肌细胞积累了更多的棕榈酰肉碱 (C16)。 38°C 培养引起的热应激会导致 CPT II 缺陷型肌细胞中的 C16 急剧增加，但在对照组中则不然。苯扎贝特降低了对照和 CPT II 缺陷型肌细胞中 C16 的含量。讨论：在本研究中，我们以高效且可重复的方式诱导 CPT II 缺陷型 hiPSC 分化为成熟肌细胞，并在肌细胞疾病模型中重现了 CPT II 缺陷型疾病表型的某些方面。这种方法解决了使用 iPSC 技术对FAO CPT II 缺陷异常进行建模的挑战，并有可能彻底改变该领域的转化研究。 (C) 2014 Elsevier Inc. 保留所有权利。
Introduction: Carnitine palmitoyltransferase II (CPT II) deficiency is an inherited disorder involving beta-oxidation of long-chain fatty acids (FAO), which leads to rhabdomyolysis and subsequent acute renal failure. The detailed mechanisms of disease pathogenesis remain unknown; however, the availability of relevant human cell types for investigation, such as skeletal muscle cells, is limited, and the development of novel disease models is required.Methods: We generated human induced pluripotent stem cells (hiPSCs) from skin fibroblasts of a Japanese patient with CPT II deficiency. Mature myocytes were differentiated from the patient-derived hiPSCs by introducing myogenic differentiation 1 (MYOD1), the master transcriptional regulator of myocyte differentiation. Using an in vitro acylcarnitine profiling assay, we investigated the effects of a hypolipidemic drug, bezafibrate, and heat stress on mitochondrial FAO in CPT II-deficient myocytes and controls.Results: CPT II-deficient myocytes accumulated more palmitoylcarnitine (C16) than did control myocytes. Heat stress, induced by incubation at 38 degrees C, leads to a robust increase of C16 in CPT II-deficient myocytes, but not in controls. Bezafibrate reduced the amount of C16 in control and CPT II-deficient myocytes.Discussion: In this study, we induced differentiation of CPT II-deficient hiPSCs into mature myocytes in a highly efficient and reproducible manner and recapitulated some aspects of the disease phenotypes of CPT II deficiency in the myocyte disease models. This approach addresses the challenges of modeling the abnormality of FAO in CPT II deficiency using iPSC technology and has the potential to revolutionize translational research in this field. (C) 2014 Elsevier Inc. All rights reserved.