Germline and Somatic Mutations in DNA Methyltransferase 3A (DNMT3A) Predispose to Pulmonary Arterial Hypertension (PAH) in Humans and Mice: Implications for Associated PAH.
Germline and Somatic Mutations in DNA Methyltransferase 3A (DNMT3A) Predispose to Pulmonary Arterial Hypertension (PAH) in Humans and Mice: Implications for Associated PAH.
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DNA 甲基转移酶 3A (DNMT3A) 的种系和体细胞突变易导致人和小鼠肺动脉高压 (PAH)：对相关 PAH 的影响。
DOI：
10.1101/2023.12.30.23300391
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Pauciulo,Michael
中科院分区：
文献类型：
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作者：
Al-Qazazi,Ruaa;Emon,IsaacM;Potus,François;Martin,AshleyY;Lima,PatriciaDA;Vlasschaert,Caitlyn;Chen,Kuang-Hueih;Wu,Danchen;Gupta,AsishDas;Noordhof,Curtis;Jefferson,Lindsay;McNaughton,AmyJM;Bick,AlexanderG;Pauciulo,Michael
背景：在10-20%的特发性肺动脉高压患者中发现突变，但在占∼45%的结缔组织病相关肺动脉高压患者中没有发现一致的突变。TET2突变是不确定潜能克隆性造血(CHIP)的原因之一，易患炎症型PAH。我们现在检查PAH中另一种芯片基因DNMT3A的突变。方法通过对1832例PAH生物库患者和7509例性别、年龄相匹配的gnomAD对照组进行全外显子测序(WES)芯片检测，评估PAH生物库受试者与对照组的DNMT3A突变发生率。然后，我们对710名PAH生物库患者的子集进行了深入的、有针对性的芯片基因小组测序，并将其中DNMT3A突变的流行率与独立的汇集对照队列(N=3645)进行了比较。在另一组80例PAH患者和41例对照组中，研究了外周血单个核细胞(PBMC)中DNMT3A mRNA的表达。最后，我们在一个条件的、造血的、DNMT3A基因敲除的小鼠模型中评估了PAH的发展。结果在WES数据集中，PAH患者的DNMT3A突变频率高于对照组(OR 2.60，95%CI：1.71~4.27)。在PAH患者中，33例存在DNMT3A变异，其中大多数为APAH(21/33)。21/33存在体细胞突变(女性：男性17：4)，12/33出现生殖系变异(女性：男性11：1)。血流动力学与有无DNMT3A突变的患者相似(mPAP=58±21对52±18毫米汞柱)；然而，有DNMT3A突变的患者对急性血管扩张剂测试没有反应。靶向面板测序发现14.6%的PAH患者存在芯片突变(104/710)，其中DNMT3A占49/104。在调整了年龄和性别的分析中，虽然单独使用DNMT3A芯片并没有显著增加(OR：1.15，0.82-1.61)，但所有的芯片突变与PAH显著相关(OR：1.40，95%CI：1.09-1.80)。患者来源的PAH-PBMC中DNMT3A的表达低于对照组。DNMT3A-/-小鼠出现自发性PAH，缺氧3周加重。DNMT3A-/-小鼠肺巨噬细胞增多，血浆IL-13升高。IL-1β抗体Canakinumab可降低DNMT3A-/-小鼠的PAH水平。结论在人类中，生殖系和获得性DNMT3A变异易患PAH。人PAH外周血单核细胞DNMT3A基因表达降低。DNMT3A的造血衰竭会引起小鼠的炎性多环芳烃。DNMT3A是一个新的APAH基因，可能成为肿瘤的生物标志物和治疗靶点。
Background Mutations are found in 10-20% of idiopathic PAH (IPAH) patients, but none are consistently identified in connective tissue disease-associated PAH (APAH), which accounts for ∼45% of PAH cases. TET2 mutations, a cause of clonal hematopoiesis of indeterminant potential (CHIP), predispose to an inflammatory type of PAH. We now examine mutations in another CHIP gene, DNMT3A , in PAH. Methods We assessed DNMT3A mutation prevalence in PAH Biobank subjects as compared with controls, first using whole exome sequencing (WES)-derived CHIP calls in 1832 PAH Biobank patients versus 7509 age-and sex-matched gnomAD controls. We then performed deep, targeted panel sequencing of CHIP genes on a subset of 710 PAH Biobank patients and compared the prevalence of DNMT3A mutations therein to an independent pooled control cohort (N = 3645). In another cohort of 80 PAH patients and 41 controls, DNMT3A mRNA expression was studied in peripheral blood mononuclear cells (PBMCs). Finally, we evaluated the development of PAH in a conditional, hematopoietic, Dnmt3a knockout mouse model. Results DNMT3A mutations were more frequent in PAH cases versus control subjects in the WES dataset (OR 2.60, 95% CI: 1.71-4.27). Among PAH patients, 33 had DNMT3A variants, most of whom had APAH (21/33). While 21/33 had somatic mutations (female:male 17:4), germline variants occurred in 12/33 (female:male 11:1). Hemodynamics were comparable with and without DNMT3A mutations (mPAP=58±21 vs. 52±18 mmHg); however, patients with DNMT3A mutations were unresponsive to acute vasodilator testing. Targeted panel sequencing identified that 14.6% of PAH patients had CHIP mutations (104/710), with DNMT3A accounting for 49/104. There was a significant association between all CHIP mutations and PAH in analyses adjusted for age and sex (OR 1.40, 95% CI: 1.09-1.80), though DNMT3A CHIP alone was not significantly enriched (OR:1.15, 0.82-1.61). DNMT3A expression was reduced in patient-derived versus control PAH-PBMCs. Spontaneous PAH developed in Dnmt3a -/- mice, and it was exacerbated by 3 weeks of hypoxia. Dnmt3a -/- mice had increased lung macrophages and elevated plasma IL-13. The IL-1β antibody canakinumab attenuated PAH in Dnmt3a -/- mice. Conclusions Germline and acquired DNMT3A variants predispose to PAH in humans. DNMT3A mRNA expression is reduced in human PAH PBMCs. Hematopoietic depletion of Dnmt3a causes inflammatory PAH in mice. DNMT3A is a novel APAH gene and may be a biomarker and therapeutic target.