Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs.
Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs.
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作者：
将体细胞重编程为患者特异性诱导多能干细胞（iPSC）使得能够对人类遗传性疾病进行体外建模，以用于病原学研究和治疗筛选。然而，使用iPSC衍生的心肌细胞（iPSC-CM）来模拟成人发作的心脏病仍然具有挑战性，这是由于关于相对不成熟的iPSC-CM完全再现成人疾病表型的能力的不确定性。致心律失常性右心室发育不良/心肌病（ARVD/C）是一种遗传性心脏病，其特征是病理性脂肪浸润和心肌细胞丢失，主要发生在右心室（RV），与危及生命的室性心律失常相关。超过50%的受影响个体具有桥粒基因突变，最常见于编码斑嗜蛋白-2的PKP2。出现ARVD/C的中位年龄为26岁。我们使用Yamanaka的方法从两名ARVD/C和PKP2突变患者的成纤维细胞中产生iPSC系。突变体PKP2 iPSC-CM在心源性疾病中表现出异常斑珠蛋白核易位和β-连环蛋白活性降低;然而，这些异常特征不足以在标准心源性疾病中重现ARVD/C的病理表型。在这里，我们表明，诱导成人样代谢能量从胚胎/糖酵解状态和异常过氧化物酶体增殖物激活受体-γ（PPAR γ）激活的基础上ARVD/C的发病机制。本研究通过共激活胚状体（EBs）中正常的过氧化物酶体增殖物激活受体α（PPAR α）依赖性代谢和异常的过氧化物酶体增殖物激活受体γ（PPAR γ）途径，在两个月内建立了有效的ARVD/C体外模型。该模型表明突变体PKP2 iPSC-CM中过度的脂肪生成和细胞凋亡。具有纯合PKP2突变的iPSC-CM也显示钙处理缺陷。我们的研究首次证明了诱导成人样代谢在使用患者特异性iPSC建立成人发病疾病模型中起着关键作用。使用该模型，我们揭示了关键的致病性见解，即成人样代谢环境中的代谢紊乱是ARVD/C病理的基础，使我们能够提出新的疾病修饰治疗策略。
Cellular reprogramming of somatic cells to patient-specific induced pluripotent stem cells (iPSCs) enables in-vitro modelling of human genetic disorders for pathogenic investigations and therapeutic screens. However, using iPSC-derived cardiomyocytes (iPSC-CMs) to model an adult-onset heart disease remains challenging due to the uncertainty regarding the ability of relatively immature iPSC-CMs to fully recapitulate adult disease phenotypes. Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) is an inherited heart disease characterized by pathological fatty infiltration and cardiomyocyte loss predominantly in the right ventricle (RV), which is associated with life-threatening ventricular arrhythmias. Over 50% of affected individuals have desmosome gene mutations, most commonly in PKP2 encoding plakophilin-2. The median age at presentation of ARVD/C is 26 years. We used Yamanaka’s methods to generate iPSC lines from fibroblasts of two patients with ARVD/C and PKP2 mutations. Mutant PKP2 iPSC-CMs demonstrate abnormal plakoglobin nuclear translocation and decreased β-catenin activity in cardiogenic conditions; yet these abnormal features are insufficient to reproduce the pathological phenotypes of ARVD/C in standard cardiogenic conditions. Here we show that induction of adult-like metabolic energetics from an embryonic/glycolytic state and abnormal peroxisome proliferator-activated receptor-gamma (PPARγ) activation underlie the pathogenesis of ARVD/C. By coactivating normal PPAR-alpha (PPARα)-dependent metabolism and abnormal PPARγ pathway in beating embryoid bodies (EBs) with defined media, we established an efficient ARVD/C in-vitro model within two months. This model manifests exaggerated lipogenesis and apoptosis in mutant PKP2 iPSC-CMs. iPSC-CMs with a homozygous PKP2 mutation also displayed calcium-handling deficits. Our study is the first to demonstrate that induction of adult-like metabolism plays a critical role in establishing an adult-onset disease model using patient-specific iPSCs. Using this model, we revealed crucial pathogenic insights that metabolic derangement in adult-like metabolic milieu underlies ARVD/C pathologies, enabling us to propose novel disease-modifying therapeutic strategies.
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影响因子：
64.5
作者：
Park IH;Arora N;Huo H;Maherali N;Ahfeldt T;Shimamura A;Lensch MW;Cowan C;Hochedlinger K;Daley GQ
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