Genetic and functional implications of an exonic TRIM55 variant in heart failure
Genetic and functional implications of an exonic TRIM55 variant in heart failure
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DOI：
10.1016/j.yjmcc.2019.12.008
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发表时间：
2020-01-01
影响因子：
5
通讯作者：
Aittokallio, Tero
中科院分区：
文献类型：
--
作者：
Heliste, Juho;Chheda, Himanshu;Aittokallio, Tero
背景资料：为了解决缺失的散发性心力衰竭的遗传性，我们筛选了新的心力衰竭相关的遗传变异在芬兰人口和功能特点的一种新的变异体在体外和体内。方法和结果：心力衰竭相关的变异体筛选基因分型阵列数据的FINRISK研究，包括994例和20,118对照。基于逻辑回归分析，选择TRIM 55中编码E140 K变体的潜在损伤性变体（rs 138811034）进行验证。在HL-1心肌细胞中，我们使用CRISPR/Cas9技术将变体引入内源性基因座中，另外从质粒过表达TRIM 55野生型或E140 K。使用全基因组RNA测序、RT-PCR和Western分析、细胞活力和细胞周期测定以及细胞表面积测量来分析功能反应。在斑马鱼胚胎中，在CRISPR介导的trim 55 a敲除或TRIM 55 WT或E140 K的质粒过表达后，使用视频显微镜测量心脏收缩力。与肌肉收缩和心脏应激相关的基因在Trim 55 E140 K/-心肌细胞中高度调节。当与WT/WT细胞相比时，变体细胞表现出活力降低、对异丙肾上腺素的显著肥大反应、p21蛋白过表达和细胞周期进展受损。在斑马鱼胚胎中，删除trim 55 a或过度表达的TRIM 55 E140 K降低心肌收缩力相比，野生型基因型或过度表达的WT TRIM 55，respectively.Conclusions：一个以前未知的TRIM 55 E140 K变异体表现出一些功能的影响，心肌细胞的功能在体外和体内。这些发现表明TRIM 55多态性在心力衰竭易感性中的新作用。
Background: To tackle the missing heritability of sporadic heart failure, we screened for novel heart failure associated genetic variants in the Finnish population and functionally characterized a novel variant in vitro and in vivo.Methods and results: Heart failure-associated variants were screened in genotyping array data of the FINRISK study, consisting of 994 cases and 20,118 controls. Based on logistic regression analysis, a potentially damaging variant in TRIM55 (rs138811034), encoding an E140K variant, was selected for validations. In HL-1 cardiomyocytes, we used CRISPR/Cas9 technology to introduce the variant in the endogenous locus, and additionally TRIM55 wildtype or E140K was overexpressed from plasmid. Functional responses were profiled using whole-genome RNA sequencing, RT-PCR and Western analyses, cell viability and cell cycle assays and cell surface area measurements. In zebrafish embryos, cardiac contractility was measured using videomicroscopy after CRISPR-mediated knockout of trim55a or plasmid overexpression of TRIM55 WT or E140K. Genes related to muscle contraction and cardiac stress were highly regulated in Trim55 E140K/- cardiomyocytes. When compared to the WT/WT cells, the variant cells demonstrated reduced viability, significant hypertrophic response to isoproterenol, p21 protein overexpression and impaired cell cycle progression. In zebrafish embryos, the deletion of trim55a or overexpression of TRIM55 E140K reduced cardiac contractility as compared to embryos with wild type genotype or overexpression of WT TRIM55, respectively.Conclusions: A previously uncharacterized TRIM55 E140K variant demonstrated a number of functional implications for cardiomyocyte functions in vitro and in vivo. These findings suggest a novel role for TRIM55 polymorphism in predisposing to heart failure.