心血管疾病,动脉粥样硬化,中风是Hutchinson-Gilford早衰综合征(HGPS)患者最常见的死亡原因。LMNA变异体c.1824C>T占HGPS病例的约90%。由于缺乏适当的体外模型,LaminA在心脏中的详细分子机制仍然难以捉摸。我们假设HGPS患者的诱导性多能干细胞(iPSC)衍生的心肌细胞(iCMC)将为研究与HGPS相关的心脏病理机制提供模型平台。为了阐明早衰素在心肌细胞中的作用,我们首先从一名去识别的HGPS患者(hPGP1,先证者)和双亲来自早衰症研究基金会的皮肤成纤维细胞(SF).通过Sanger测序和限制性片段长度多态性,酶EciI,靶向LaminA,我们将hPGP1-SF表征为LMNA变体c.1824C>T的杂合突变体。我们进行了LMNA外显子11亚硫酸氢盐测序以分析早衰细胞的甲基化状态.此外,我们将三个SF重新编程为iPSCs并将它们分化为iCMCs,在第7天遭到殴打。通过粒子图像测速分析,通过qRT-PCR和Westernblot,我们发现hPGP1-iCMCs具有不规则的收缩功能,心脏特异性基因和蛋白表达降低.与正常的iCMC相比,我们的早衰患者来源的iCMC在功能和结构上都有缺陷。该体外模型将有助于阐明LaminA在心脏疾病中的作用以及与早衰相关的心脏病理机制。它为研究人员提供了一个新的平台,以研究与早衰相关的心脏病的新治疗方法。
Cardiovascular diseases, atherosclerosis, and strokes are the most common causes of death in patients with Hutchinson-Gilford progeria syndrome (HGPS). The LMNA variant c.1824C > T accounts for ~ 90% of HGPS cases. The detailed molecular mechanisms of Lamin A in the heart remain elusive due to the lack of appropriate in vitro models. We hypothesize that HGPS patient\'s induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iCMCs) will provide a model platform to study the cardio-pathologic mechanisms associated with HGPS. To elucidate the effects of progerin in cardiomyocytes, we first obtained skin fibroblasts (SFs) from a de-identified HGPS patient (hPGP1, proband) and both parents from the Progeria Research Foundation. Through Sanger sequencing and restriction fragment length polymorphism, with the enzyme EciI, targeting Lamin A, we characterized hPGP1-SFs as heterozygous mutants for the LMNA variant c.1824 C > T. Additionally, we performed LMNA exon 11 bisulfite sequencing to analyze the methylation status of the progeria cells. Furthermore, we reprogrammed the three SFs into iPSCs and differentiated them into iCMCs, which gained a beating on day 7. Through particle image velocimetry analysis, we found that hPGP1-iCMCs had an irregular contractile function and decreased cardiac-specific gene and protein expressions by qRT-PCR and Western blot. Our progeria-patient-derived iCMCs were found to be functionally and structurally defective when compared to normal iCMCs. This in vitro model will help in elucidating the role of Lamin A in cardiac diseases and the cardio-pathologic mechanisms associated with progeria. It provides a new platform for researchers to study novel treatment approaches for progeria-associated cardiac diseases.