Successful knock-in of Hypertrophic Cardiomyopathy-mutation R723G into the MYH7 gene mimics HCM pathology in pigs.
Successful knock-in of Hypertrophic Cardiomyopathy-mutation R723G into the MYH7 gene mimics HCM pathology in pigs.
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DOI：
10.1038/s41598-018-22936-z
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发表时间：
2018-03-19
影响因子：
4.6
通讯作者：
Brenner B
中科院分区：
文献类型：
--
作者：
Montag J;Petersen B;Flögel AK;Becker E;Lucas-Hahn A;Cost GJ;Mühlfeld C;Kraft T;Niemann H;Brenner B
家族性肥厚型心肌病（Familial Hypertrophic Cardiomyopathy，HCM）是最常见的遗传性心脏病。约30%的患者是编码β-肌球蛋白重链（MyHC）的MYH 7基因突变的杂合型。HCM的特征是心肌细胞紊乱和左心室肥大，症状从轻微的心律失常到心脏性猝死或心力衰竭。为了深入了解疾病病因学的潜在机制，我们的目标是产生具有HCM突变的基因组编辑的猪。我们使用TALEN介导的基因组编辑，并成功地将HCM点突变R723 G引入猪成纤维细胞的MYH 7基因中，随后克隆了HCM突变R723 G杂合的猪。在R723 G猪中未确定脱靶效应。令人惊讶的是，动物在产后24小时内死亡，可能是由于心力衰竭，如左心室中α/β-MyHC比率的变化所指示的。最有趣的是，新生猪表现出HCM的特征，包括轻度肌细胞紊乱、畸形核和MYH 7过表达。在新生R723 G仔猪中发现HCM特异性病理学表明该疾病的发病非常早，并强调了新型大型动物模型对于研究人类心脏疾病的致病机制和长期进展的重要性。
Familial Hypertrophic Cardiomyopathy (HCM) is the most common inherited cardiac disease. About 30% of the patients are heterozygous for mutations in the MYH7 gene encoding the ß-myosin heavy chain (MyHC). Hallmarks of HCM are cardiomyocyte disarray and hypertrophy of the left ventricle, the symptoms range from slight arrhythmias to sudden cardiac death or heart failure. To gain insight into the underlying mechanisms of the diseases’ etiology we aimed to generate genome edited pigs with an HCM-mutation. We used TALEN-mediated genome editing and successfully introduced the HCM-point mutation R723G into the MYH7 gene of porcine fibroblasts and subsequently cloned pigs that were heterozygous for the HCM-mutation R723G. No off-target effects were determined in the R723G-pigs. Surprisingly, the animals died within 24 h post partem, probably due to heart failure as indicated by a shift in the a/ß-MyHC ratio in the left ventricle. Most interestingly, the neonatal pigs displayed features of HCM, including mild myocyte disarray, malformed nuclei, and MYH7-overexpression. The finding of HCM-specific pathology in neonatal R723G-piglets suggests a very early onset of the disease and highlights the importance of novel large animal models for studying causative mechanisms and long-term progression of human cardiac diseases.
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