Endothelial Injury, BMPR2 Dysfunction and Macrophage Activation Cause EndMT and PAH

内皮损伤、BMPR2 功能障碍和巨噬细胞激活导致 EndMT 和 PAH

• 批准号: 9367124
• 负责人: Mark Robert Nicolls
• 金额: $ 77.64万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9367124
• 关键词: Anabolism; Animals; Apoptosis; Arachidonate 5-Lipoxygenase; Arterial Disorder; Autoimmune Process; Autoimmunity; Biological Models; Biological Response Modifiers; Biology; Blood Vessels; Cells; Clinical Trials; Confocal Microscopy; Coupled; Data; Development; Disease; Drug Targeting; Endogenous Retroviruses; Endothelial Cells; Evolution; Flow Cytometry; Functional disorder; Gene Expression; Gene Expression Profile; Genetic; Genetic Predisposition to Disease; Genetic Transcription; Genetic Translation; Genotype; Human; Hypoxia; IL6 gene; Immune; Immunologic Factors; Immunotherapy; Impairment; Inflammation; Inflammatory; Inflammatory Response; Injury; Intravenous; Knowledge; Laboratories; Lead; Leukotriene B4; Leukotriene Production; Leukotrienes; Link; Lung; Macrophage Activation; Maps; Mediating; Mesenchymal; Microfluidics; Modeling; Mus; Natural Immunity; Oxidants; Pathway interactions; Patients; Play; Population; Process; Production; Proteins; Pulmonary Hypertension; Pulmonary Pathology; Rattus; Recombinants; Recruitment Activity; Recurrence; Resistance; Retroviridae Infections; Rodent Model; Role; Signal Transduction; Stimulus; Stress; TP53 gene; Testing; Time; Transferase; Transgenic Organisms; Vascular Diseases; Vascular remodeling; Virus Diseases; Work; bone loss; bone morphogenetic protein receptors; cell injury; cell transformation; cytokine; dUTP pyrophosphatase; endotracheal; injured; macrophage; monocyte; mutant; pulmonary arterial hypertension; pulmonary artery endothelial cell; receptor; response; success; transcriptome sequencing

A considerable body of knowledge indicating that inflammation and immune dysregulation play a pivotal role in the development of pulmonary arterial hypertension (PAH) have now led to the first immunotherapy trials for this condition. However the success of this approach requires that we determine how the aberrant immune/inflammatory response to pulmonary artery endothelial cell (PAEC) injury develops and contributes to the evolution of the occlusive arteriopathy, that is characterized by apoptosis, apoptosis resistance and endothelial mesenchymal transition (EndMT). It is also important to relate these features to dysfunction of a key genetic modifier of PAH, bone morphogenetic protein receptor (BMPR)2. This proposal brings unique focus to two key innate immune mediators that are highly expressed in perivascular macrophages (MØs) from patients with PAH and that mount a coordinated attack on the pulmonary vasculature: the leukotriene, LTB4, and the endogenous retrovirus, HERV-K, an increasingly recognized cause of autoimmune injury. The overall hypothesis is injured PAECs with dysfunctional BMPR2 signaling recruit and activate MØs that amplify LTB4 and HERV-K; these immune factors work in concert to sustain inflammation and promote severe PAH by apoptosis and EndMT. Aim 1 evaluates how LTB4 may be autonomously produced by injured PAECs, which in turn activate MØs to stimulate further LTB4 biosynthesis to cause PAEC apoptosis and EndMT when BMPR2 signaling is impaired. This aim also explores how amplification of endogenous HERV-K may further stimulate LTB4 biosynthesis and how LTB4 can induce HERV-K expression in monocytes. Single cell RNA Seq will genotype PAEC populations that are fated for apoptosis or EndMT in response to LTB4 to delineate gene expression changes that mark the transition to apoptosis resistance and EndMT. Aim 2 evaluates whether PAECs from PAH patients including those with reduced BMPR2 function, secrete factors in response to oxidant or inflammatory injury, that amplify HERV-K expression in monocytes. This aim also considers the additive effects of loss of BMPR2 in monocytes, on amplification of HERV-K and whether transcriptional and translational mechanisms are involved in this process. Also to be investigated is whether the secreted HERV-K product, dUTPase, promotes EndMT in PAEC with BMPR2 dysfunction that are primed to undergo transformation. Single cell RNA Seq analyses will elucidate gene expression changes induced by HERV-K in the PAEC evolution to EndMT. Aim 3 investigates the vulnerability of BMPR2-deficient animals to develop severe PH and EndMT using a mouse with loss of BMPR2 in fate mapped EC, and a new transgenic rat PH model with BMPR2 haploinsufficiency. The animals are treated with endotracheal instillation of Ad-5LO to generate high LTB4 levels or recombinant HERV-K dUTPase given intravenously. Strategies to reverse EndMT and severe PH will be tested that have translational relevance to the fatal obliterative vasculopathy in PAH patients.

大量知识表明注射和免疫失调在 现在，肺动脉高压（PAH）的发展已导致首次进行免疫疗法试验 这种情况。但是，这种方法的成功要求我们确定异常 对肺部动脉内皮细胞（PAEC）损伤发生的免疫/炎症反应，并有助于 闭塞性动脉疾病的演变，其特征是凋亡，凋亡耐药性和 内皮间充质转变（ENDMT）。将这些功能与功能障碍联系起来也很重要 PAH，骨形态发生蛋白受体（BMPR）2的关键遗传修饰剂2。该建议带来了独特的 专注于两个主要的先天免疫介质，这些介体在血管周围巨噬细胞（Møs）中高度表达 患有PAH的患者并对肺脉管系统进行了协调的攻击：白细胞三烯，LTB4， 以及内源性逆转录病毒HERV-K，这是自身免疫损伤的日益认识的原因。总体 假设是受伤的PAEC，具有功能失调的BMPR2信号募集并激活Møs，以扩大LTB4 和Herv-k；这些免疫因素协同工作以维持注射和促进严重的PAH 凋亡和终结。 AIM 1评估LTB4如何由受伤的PAEC自主生产，这在 转动激活Møs以刺激进一步的LTB4生物合成，以引起BMPR2时引起PAEC凋亡和EndMT 信号传导受损。该目标还探讨了内源性HERV-K的扩增如何进一步刺激 LTB4生物合成以及LTB4如何诱导单核细胞中的HERV-K表达。单细胞RNA SEQ将 响应于LTB4的凋亡或EndMT的基因型PAEC种群以描绘基因 表达变化，标志着向凋亡耐药性和终结物的过渡。 AIM 2评估是否 来自PAH患者的PAEC，包括BMPR2功能降低的患者，响应的秘密因素 氧化剂或炎症性损伤，可以扩增单核细胞中的HERV-K表达。这个目标也考虑 单核细胞中BMPR2丢失的加性影响，对HERV-K的扩增以及转录和是否转录 翻译机制参与了这一过程。还需要调查是分泌的赫尔夫-K dutpase的产品促进了paec的Endmt，它具有BMPR2功能障碍，该功能障碍 转型。单细胞RNA SEQ分析将阐明HERV-K在 PAEC演变为Endmt。 AIM 3调查了BMPR2缺陷动物发展的脆弱性 使用小鼠使用脂肪映射EC中的BMPR2的小鼠和新的转基因大鼠pH的严重pH和EndMT 具有BMPR2单倍宽度的模型。将动物用气管插入AD-5LO进行处理 产生高LTB4水平或重组HERV-K dutpase静脉注射。反向端的策略 将测试严重的pH值，这些pH已转化为与PAH中致命的闭塞性血管病有关 患者。