Novel germline mutation KMT2A G3131S confers genetic susceptibility to familial myeloproliferative neoplasms
Novel germline mutation KMT2A G3131S confers genetic susceptibility to familial myeloproliferative neoplasms
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新种系突变 KMT2A G3131S 赋予家族性骨髓增生性肿瘤遗传易感性
DOI：
10.1007/s00277-021-04562-4
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发表时间：
2021-07
影响因子：
3.5
通讯作者：
Ji Li
中科院分区：
文献类型：
--
作者：
Hongling Peng;Zhao Cheng;Yi Chen;Guangsen Zhang;Sisi Xie;Heng Li;Xiang Xiao;Zefang Wu;Wang Li;Xian Jiang;Le Yin;Ji Li
本研究分析了一个家族性骨髓增生性肿瘤（mpn）的临床和遗传特征。进行全外显子组测序，鉴定出赖氨酸甲基转移酶2A （KMT2A，也称为MLL1）种系杂合突变G3131S （c.9391G > a, p.Gly3131Ser, rs150804738）。采集8名家庭成员、120名健康供体（体细胞DNA）和30名健康供体（种系DNA）的体细胞DNA和种系DNA。使用Sanger测序对KMT2A G3131S突变进行分析。先证者（II-1）、其妹妹（患者II-2）和家庭成员II-3和III-1（体细胞DNA和种系DNA） 4人检测KMT2A G3131S突变阳性。我们没有在健康供体（体细胞DNA和种系DNA）中观察到KMT2A G3131S突变，表明这不是一个SNP。对KMT2A G3131S的生物信息学分析表明，该突变可能引起蛋白质结构的改变。为了进一步阐明KMT2A G3131S的功能，我们利用CRISPR-Cas9技术生成了KMT2A G3131S杂合的K562细胞系。分析KMT2A G3131S突变体K562细胞的集落形成能力、细胞凋亡和细胞周期。结果表明，KMT2A G3131S突变体K562细胞增殖能力和集落形成能力增强。利用流式细胞术进行免疫分型，分析基因编辑后KMT2A G3131S突变体K562细胞的表面标记物表达。KMT2A G3131S突变体K562细胞中CD11b显著升高，CD61和CD235a轻度升高，提示KMT2A G3131S突变体可引起骨髓增殖增加。May-Giemsa染色显示KMT2A G3131S突变体K562细胞的形态学变化与流式细胞术分析一致。为了验证哪些下游基因受到KMT2A G3131S突变体的影响，我们采用实时荧光定量PCR方法评估了之前报道的KMT2A相关基因的表达，发现C-MYB的表达显著降低。Western blotting检测Kmt2a和C-myb蛋白的表达，结果显示，在Kmt2a G3131S突变体K562细胞中，C-myb的表达降低。提示KMT2A G3131S可影响K562细胞的骨髓增殖，降低C-myb的表达。综上所述，KMT2A G3131S可能是家族性MPN的一个新的遗传易感基因。
The current study analyzed the clinical and genetic characteristics of a family with familial myeloproliferative neoplasms (MPNs). Whole-exome sequencing was conducted, and a germline heterozygous mutation in lysine methyltransferase 2A (KMT2A, also known as MLL1), G3131S (c.9391G > A, p.Gly3131Ser, rs150804738), was identified. Somatic DNA and germline DNA were collected from 8 family members, 120 healthy donors (somatic DNA), and 30 healthy donors (germline DNA). Using Sanger sequencing, the KMT2A G3131S mutation was analyzed. Four individuals, the proband (II-1), his sister (patient II-2), and family members II-3 and III-1 (somatic DNA and germline DNA), tested positive for the KMT2A G3131S mutation. We did not observe the KMT2A G3131S mutation in healthy donors (somatic DNA and germline DNA), indicating that this is not a SNP. Bioinformatics analysis of KMT2A G3131S suggested that protein structure changes could be caused by this mutation. To further elucidate the function of KMT2A G3131S, the CRISPR-Cas9 technique was applied to generate a KMT2A G3131S heterozygous K562 cell line. The colony formation potency, apoptosis, and cell cycle of KMT2A G3131S mutant K562 cells were analyzed. The results demonstrated that KMT2A G3131S mutant K562 cells showed increased proliferation and colony formation ability. Immunophenotyping was performed using flow cytometry to analyze the surface marker expression of gene-edited KMT2A G3131S mutant K562 cells. A significant increase in CD11b and mild increases in CD61 and CD235a were observed in KMT2A G3131S mutant K562 cells, suggesting that the KMT2A G3131S mutant could cause an increase in myeloproliferation. May-Giemsa staining showed that the morphological changes in KMT2A G3131S mutant K562 cells were consistent with the flow cytometry analysis. To verify which downstream genes were affected by the KMT2A G3131S mutant, we performed real-time PCR to evaluate the expression of previously reported KMT2A-related genes and found that C-MYB expression was significantly decreased. Western blotting was applied to investigate the expression of Kmt2a and C-myb proteins, and the results showed that in KMT2A G3131S mutant K562 cells, the expression of C-myb was decreased. Our findings suggested that KMT2A G3131S could affect the myeloproliferation of K562 cells and decrease C-myb expression. In conclusion, KMT2A G3131S could be considered a novel genetic susceptibility gene in familial MPN.
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影响因子：
11.2
作者：
Chen J;Santillan DA;Koonce M;Wei W;Luo R;Thirman MJ;Zeleznik-Le NJ;Diaz MO
通讯作者：
Diaz MO
DOI：
10.1158/1078-0432.ccr-14-2300
发表时间：
2015-03-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Richmond J;Carol H;Evans K;High L;Mendomo A;Robbins A;Meyer C;Venn NC;Marschalek R;Henderson M;Sutton R;Kurmasheva RT;Kees UR;Houghton PJ;Smith MA;Lock RB
通讯作者：
Lock RB
影响因子：
2.3
作者：
Malak, Sandra;Labopin, Myriam;Najman, Albert
通讯作者：
Najman, Albert
影响因子：
16
作者：
Milne, TA;Briggs, SD;Hess, JL
通讯作者：
Hess, JL
影响因子：
12.3
作者：
Goldin LR;Björkholm M;Kristinsson SY;Samuelsson J;Landgren O
通讯作者：
Landgren O