Pienkos Shaun, Gallego Natalia, Condon David F, Cruz-Utrilla Alejandro, Ochoa Nuria, Nevado Julián, Arias Pedro, Agarwal Stuti, Patel Hiral, Chakraborty Ananya, Lapunzina Pablo, Escribano Pilar, Tenorio-Castaño Jair, de Jesús Pérez Vinicio A
Division of Pulmonary and Critical Care Medicine and Department of Medicine, Stanford University, Stanford, CA, United States.
Medical and Molecular Genetics Institute (INGEMM), IdiPaz, Hospital Universitario La Paz, Madrid, Spain.
Front Med (Lausanne). 2021 Apr 30;8:625763. doi: 10.3389/fmed.2021.625763. eCollection 2021.
Pulmonary arterial hypertension (PAH) is a rare disease characterized by pulmonary vascular remodeling and right heart failure. Specific genetic variants increase the incidence of PAH in carriers with a family history of PAH, those who suffer from certain medical conditions, and even those with no apparent risk factors. Inflammation and immune dysregulation are related to vascular remodeling in PAH, but whether genetic susceptibility modifies the PAH immune response is unclear. and encode for immunomodulatory proteins that regulate NF-κB activation, a transcription factor complex associated with inflammation and vascular remodeling in PAH. Two unrelated families with PAH cases underwent whole-exome sequencing (WES). A custom pipeline for variant prioritization was carried out to obtain candidate variants. To determine the impact of TNIP2 and TRAF2 in cell proliferation, we performed an MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] assay on healthy lung pericytes transfected with siRNA specific for each gene. To measure the effect of loss of TNIP2 and TRAF2 on NF-kappa-beta (NF-κB) activity, we measured levels of Phospho-p65-NF-κB in siRNA-transfected pericytes using western immunoblotting. We discovered a novel missense variant in the gene in two affected individuals from the same family. The two patients had a complex form of PAH with interatrial communication and scleroderma. In the second family, WES of the proband with PAH and primary biliary cirrhosis revealed a protein-truncating variant in the . The knockdown of TNIP2 and TRAF2 increased NF-κB activity in healthy lung pericytes, which correlated with a significant increase in proliferation over 24 h. We have identified two rare novel variants in and using WES. We speculate that loss of function in these genes promotes pulmonary vascular remodeling by allowing overactivation of the NF-κB signaling activity. Our findings support a role for WES in helping identify novel genetic variants associated with dysfunctional immune response in PAH.
肺动脉高压（PAH）是一种罕见疾病，其特征为肺血管重塑和右心衰竭。特定的基因变异会增加有PAH家族病史的携带者、患有某些疾病的人群甚至无明显风险因素者患PAH的几率。炎症和免疫失调与PAH中的血管重塑有关，但基因易感性是否会改变PAH免疫反应尚不清楚。TNIP2和TRAF2编码免疫调节蛋白，这些蛋白调节NF-κB激活，NF-κB是一种与PAH中的炎症和血管重塑相关的转录因子复合体。两个有PAH病例的无关家族接受了全外显子组测序（WES）。实施了一个用于变异体优先级排序的定制流程以获得候选变异体。为了确定TNIP2和TRAF2对细胞增殖的影响，我们对用针对每个基因的小干扰RNA（siRNA）转染的健康肺周细胞进行了MTS[3-(4,5-二甲基噻唑-2-基)-5-(3-羧基甲氧基苯基)-2-(4-磺基苯基)-2H-四唑]检测。为了测量TNIP2和TRAF2缺失对NF-κB活性的影响，我们使用western免疫印迹法测量了siRNA转染的周细胞中磷酸化p65-NF-κB的水平。我们在来自同一家族的两名受影响个体的TNIP2基因中发现了一个新的错义变异。这两名患者患有伴有房间隔交通和硬皮病的复杂形式的PAH。在第二个家族中，对患有PAH和原发性胆汁性肝硬化的先证者进行的WES揭示了TRAF2基因中的一个蛋白质截短变异。TNIP2和TRAF2的敲低增加了健康肺周细胞中的NF-κB活性，这与24小时内增殖的显著增加相关。我们使用WES在TNIP2和TRAF2中鉴定出两个罕见的新变异。我们推测这些基因的功能丧失通过允许NF-κB信号活性过度激活而促进肺血管重塑。我们的研究结果支持WES在帮助识别与PAH中功能失调的免疫反应相关的新基因变异方面的作用。
