心肌致密化不全新致病基因的筛选及其表达特征变化

Screening for new causative genes and their expression profiles in patients with left ventricular noncompaction

目的筛选心肌致密化不全(left ventricular noncompaction,LVNC)的致病基因并进行功能验证,为研究发病机制、遗传咨询和早期干预提供依据。方法收集自2018年1月至2020年12月于重庆医科大学附属儿童医院门诊就诊的3个LVNC家系的血液样本进行全外显子测序(whole-exome sequencing,WES),得到3个家系均筛选出的可能致病基因,同时分别构建LVNC患者和正常人来源的诱导多能干细胞-心肌细胞(induced pluripotent stem cells-cardiomyocytes,iPSC-CMs)模型,收取不同时间点的细胞,采用实时荧光定量PCR(quantitative real-time PCR,qPCR)对上述基因进行验证,观察它们在诱导分化过程中的表达特征变化。结果WES筛选出3个LVNC家系共同的可能致病基因14个,qPCR检测结果提示这些基因在不同的时间点(0、10、20、30 d)表达量发生动态改变,并且在正常组及LVNC患者组间其表达量存在明显差异。结论3个LVNC家系通过WES筛选出的14个共同的可能致病基因,涉及影响细胞骨架、细胞极性、钙离子钾离子调节及能量代谢等方面。

Objective To screen the pathogenic genes in the patients with left ventricular noncompaction(LVNC)and validate their functions in order to provide references for pathogenesis,genetic counseling and early intervention of the disease.Methods Blood samples from 3 LVNC families attending the outpatient clinic of our hospital from January 2018 to December 2020 were collected for whole-exome sequencing(WES)to obtain the possible pathogenic genes screened in all 3 families.Induced pluripotent stem cells-cardiomyocytes(IPSCs-CMs)were derived and constructed separately from LVNC patients and normal individuals.The cells were collected at different time points and validated by quantitative real-time PCR(qPCR)to observe the expression profiles of above obtained genes during induction and differentiation.Results WES screened 14 possible pathogenic genes commonly expressed in the 3 LVNC families.qPCR results suggested that the expression of these genes changed dynamically at different time points(0,10,20 and 30 d),and their expression differed significantly from the normal and LVNC patient groups.Conclusion There were 14 common possible pathogenic genes screened by WES in the 3 LVNC families,and they are involved in affecting cytoskeleton,cell polarity,calcium and potassium ion regulation and energy metabolism.