Compound heterozygous mutation of theASXL3gene causes autosomal recessive congenital heart disease
Compound heterozygous mutation of theASXL3gene causes autosomal recessive congenital heart disease
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ASXL3基因复合杂合突变导致常染色体隐性遗传先天性心脏病
DOI：
10.1007/s00439-020-02200-z
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发表时间：
2020-07-21
期刊：
影响因子：
5.3
通讯作者：
Liao, Can
中科院分区：
文献类型：
--
作者：
Fu, Fang;Li, Ru;Liao, Can
利用全外显子组测序（WES）揭示了asxl3基因中与冠心病相关的一种新的复合杂合突变。对另外122名冠心病患者进行Sanger测序，以确定asxl3基因的另一个复合杂合突变。MTS法和流式细胞术检测细胞凋亡。通过苏木精-伊红（HE）、马松三色和超声扫描鉴定心脏结构。进行RNA测序以鉴定一系列差异表达的mrna。采用实时荧光定量PCR和western blotting分别检测mRNA和蛋白的表达。在asxl3基因中发现了与冠心病相关的复合杂合突变c.2168C > G （p.Pro723Arg）和c.5449C > G （p.Pro1817Ala）。这种复合杂合突变在HL-1细胞中的过度表达导致细胞凋亡增加和细胞活力降低。此外，它还影响小鼠的心脏结构和纤维化。asxl3复合杂合突变的c.2168C > G （p.Pro723Arg）和c.5449C > G （p.Pro1817Ala）小鼠与野生型小鼠之间有126个mrna下调，117个mrna上调。Ezh2、Slc6a4和socs3可以通过蛋白与asxl3相互作用，均上调。通过进一步筛选122例冠心病患者，在asxl3基因中发现了另一个复合杂合突变c.3526C > T （p.Arg1176Trp）和c. 4634a > G （p.Asp1548Gly）。asxl3基因在心脏发育中很重要，可能通过影响与细胞凋亡和细胞增殖相关的mrna的表达来发挥这种影响。
To explore mutations in the additional sex combs-like 3 (ASXL3) gene in two Chinese families with congenital heart disease (CHD). Whole-exome sequencing (WES) was used to reveal a novel compound heterozygous mutation in theASXL3gene that was associated with CHD. Sanger sequencing of a further 122 CHD patients was used to determine an additional compound heterozygous mutation in theASXL3gene. Cell apoptosis was examined by MTS assay and flow cytometry. The cardiac structure was identified via hematoxylin-eosin (HE), Masson's trichrome, and ultrasound scanning. RNA sequencing was performed to identify a series of differentially expressed mRNAs. The mRNA and protein expressions were identified by quantitative real-time PCR and western blotting, respectively. A compound heterozygous mutation c.2168C > G (p.Pro723Arg) and c.5449C > G (p.Pro1817Ala) in theASXL3gene associated with CHD was identified. Overexpression of this compound heterozygous mutation in HL-1 cells resulted in increased apoptosis and reduced cell viability. Moreover, it affected cardiac structure and fibrosis in mice. There were 126 downregulated mRNAs and 117 upregulated mRNAs between theASXL3compound heterozygous mutation c.2168C > G (p.Pro723Arg) and c.5449C > G (p.Pro1817Ala) mice and wild-type mice.Ezh2,Slc6a4, andSocs3, which could interact withASXL3through proteins, were all upregulated. Another compound heterozygous mutation c.3526C > T (p.Arg1176Trp) and c.4643A > G (p.Asp1548Gly) in theASXL3gene was identified by screening a further 122 patients with CHD. TheASXL3gene is important in cardiac development and may exert this influence by affecting the expression of mRNAs associated with cell apoptosis and cell proliferation.