Role of Chromosome Y gene, Uty, in protecting against Pulmonary Hypertension

摘要 摘要肺动脉高压(PAH)是一种以肺动脉增多为特征的慢性肺部疾病。 压力导致右室肥大、右室衰竭和死亡。多环芳烃的发生率很高 女性患者较高(4：1)。虽然之前研究多环芳烃的性别差异主要集中在 广泛研究性腺激素的作用，特别是雌激素，我们是第一个研究性腺激素的作用的实验室 多环芳烃的性染色体。我们最近发表的使用创新的鼠标模型的工作演示在 缺乏性激素，Y染色体(CHRY)对实验性肺动脉高压有保护作用 (PH)，表明编码在CHRY上的基因(S)对PH具有保护作用。只有4个基因在CHRY上表达 肺UTY、Kdm5d、Eif2s3y、Ddx3y。我们的初步数据确定UTY是首选候选基因 负责PH值的CHRY防护工作。此外，我们还证明UTY在男性中的表达减少 PAH患者和多个PH动物模型。我们对PH野生型(WT)肺组织的RNAseq分析 UTY-KD雄性小鼠发现了一些有希望的靶点，包括促炎细胞因子Cxcl9和 Cxcl10。我们的初步数据显示，UTY与Cxcl9/10在肺巨噬细胞中共定位，并表达 Cxcl9/10在UTY全球KO小鼠骨髓来源的巨噬细胞中显著增加 与WT相比。更重要的是，我们的初步研究表明，阻断共享的Cxcl9/10受体Cxcr3， 应用AMG487可降低雌性大鼠肺高压的严重程度。我们的生物信息学分析还发现 内皮素-2(ET-2)在UTY-KD时肺组织表达上调。ET-2的作用目前尚不清楚。 啊哈。我们首次表明，ET-2表达增加可能通过抑制PH恶化而起作用 促进肺血管生成和促进SMC增殖。我们的工作假设是：(1)CHRY基因UTY 防止PH的发展；(2)UTY的缺失或缺失会通过增加 Cxcl9/10和ET-2的表达导致血管内皮细胞死亡、SMC增殖和病理改变 血管生成；(3)阻断Cxcl9/10或联合阻断ET-2活性可降低PH的严重程度。 以一种针对性别的方式。目的1.检查UTY在肺中的敲除是否在存在和 缺乏激素，取消CHRY对实验性PH的保护作用；目的2.研究 UTY/Cxcl9/10和UTY/ET-2轴在PH发病机制中的作用；目的3.确定是否阻断 UTY下游基因Cxcl9/10单独或联合阻断ET-2的活性通过以下途径挽救PH的发生 减少雄性和雌性大鼠血管内皮细胞凋亡和SMC增殖，促进血管生成。

ABSTRACT Pulmonary arterial hypertension (PAH) is a chronic lung disease characterized by increased pulmonary artery pressure leading to right ventricular (RV) hypertrophy, RV failure and death. The incidence of PAH is much higher in female patients (4:1 ratio). While previous studies investigating sex differences in PAH have focused extensively on the role of gonadal hormones, in particular estrogen, we are the first lab to investigate the role of sex chromosomes in PAH. Our recent published work using innovative mouse models demonstrated in the absence of sex hormones, the Y chromosome (ChrY) protects against experimental pulmonary hypertension (PH), indicating that gene(s) encoded on ChrY can protect against PH. Only 4 genes on ChrY are expressed in the lungs Uty, Kdm5d, Eif2s3y, and Ddx3y. Our preliminary data identified Uty as the top candidate gene responsible for ChrY protection against PH. Additionally, we demonstrate that Uty expression is reduced in male patients with PAH and multiple animal models of PH. Our RNAseq analysis on the lungs of PH wildtype (WT) and Uty-KD male mice revealed a few promising targets including the proinflammatory cytokines Cxcl9 and Cxcl10. Our preliminary data shows Uty co-localizes with Cxcl9/10 in lung macrophages, and expression of Cxcl9/10 is significantly increased in bone marrow derived macrophages isolated from Uty global KO mice compared to WT. More importantly, our pilot study shows that blocking the shared Cxcl9/10 receptor, Cxcr3, using AMG487 can reduce PH severity in female rats with PH. Our bioinformatics analysis also identified Endothelin-2 (ET-2) is up-regulated in the lung as a result of Uty-KD. The role of ET-2 is currently unknown in PAH. For the first time, we show that increased ET-2 expression may contribute to worsening PH by inhibiting angiogenesis and promoting SMC proliferation in the lung. Our working hypotheses are: (1) ChrY gene Uty protects against PH development; (2) loss or absence of Uty results in more severe PH through increased expression of Cxcl9/10 and ET-2 resulting in vascular EC death, SMC proliferation and pathological angiogenesis; and (3) Blocking Cxcl9/10 alone or together with blocking ET-2 activity reduces the severity of PH in a sex-specific manner. Aim 1. To examine whether knockdown of Uty in the lungs, in the presence and absence of hormones, abolishes the protective role of ChrY in experimental PH; Aim 2. Investigate the mechanistic role of the Uty/Cxcl9/10 and Uty/ET-2 axes in PH pathogenesis; Aim 3. Determine if blocking the activity of downstream Uty genes Cxcl9/10 alone or together with blocking ET-2 rescues PH development by reducing EC apoptosis and SMC proliferation and promoting angiogenesis in male and female rats.