Pavlova Sophia V, Shulgina Angelina E, Minina Julia M, Zakian Suren M, Dementyeva Elena V
Federal Research Centre Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia.
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia.
Int J Mol Sci. 2024 Nov 30;25(23):12900. doi: 10.3390/ijms252312900.
The clinical significance of numerous cardiovascular gene variants remains to be determined. CRISPR/Cas9 allows for the introduction and/or correction of a certain variant in induced pluripotent stem cells (iPSCs). The resulting isogenic iPSC lines can be differentiated into cardiomyocytes and used as a platform to assess the pathogenicity of the variant. In this study, isogenic iPSC lines were generated for a variant of unknown significance found previously in a patient with hypertrophic cardiomyopathy (HCM), p.N515del (c.1543_1545delAAC) in . The deletion was corrected with CRISPR/Cas9 in the patient-specific iPSCs. The iPSC lines with the corrected deletion in maintained pluripotency and a normal karyotype and showed no off-target CRISPR/Cas9 activity. The isogenic iPSC lines, together with isogenic iPSC lines generated earlier via introducing the p.N515del (c.1543_1545delAAC) variant in of iPSCs of a healthy donor, were differentiated into cardiomyocytes. The cardiomyocytes derived from both panels of the isogenic iPSCs showed an increased size in the presence of the deletion in , which is one of the HCM traits at the cellular level. This finding indicates the effectiveness of these iPSC lines for studying the impact of the variant on HCM development.
众多心血管基因变异的临床意义仍有待确定。CRISPR/Cas9技术可在诱导多能干细胞（iPSC）中引入和/或校正特定变异。由此产生的同基因iPSC系可分化为心肌细胞，并用作评估该变异致病性的平台。在本研究中，针对先前在一名肥厚型心肌病（HCM）患者中发现的意义不明的变异p.N515del（c.1543_1545delAAC）构建了同基因iPSC系。利用CRISPR/Cas9在患者特异性iPSC中校正了该缺失。校正缺失后的iPSC系保持了多能性和正常核型，且未显示出脱靶的CRISPR/Cas9活性。将这些同基因iPSC系与先前通过在健康供体的iPSC的中引入p.N515del（c.1543_1545delAAC）变异而产生的同基因iPSC系一起分化为心肌细胞。来自这两组同基因iPSC的心肌细胞在存在缺失时均显示出增大的尺寸，这是细胞水平上HCM的特征之一。这一发现表明这些iPSC系在研究该变异对HCM发展的影响方面是有效的。
