Role of intestinal inflammation in oxidized lipid induced pulmonary hypertension

肠道炎症在氧化脂质诱导的肺动脉高压中的作用

• 批准号: 10381356
• 负责人: Mansoureh Eghbali
• 金额: $ 61.11万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-10 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10381356
• 关键词: Apoptosis; Bioinformatics; Blood Vessels; C57BL/6 Mouse; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; CXCL10 gene; Cells; Cessation of life; Chronic lung disease; Conditioned Culture Media; Data; Development; Diet; Dietary Supplementation; Disease; Endothelial Cells; Epithelial; Epithelial Cells; Experimental Designs; Fatty Acids; Feedback; Functional disorder; Gene Chips; Genes; Human; Hydroxyeicosatetraenoic Acids; Hypertension; Immune; Immune system; Immunohistochemistry; Inflammation; Inflammatory; Interferon Type II; Interferons; Intestines; Lamina Propria; Ligands; Lipoxygenase; Lung; Lung Transplantation; Mediating; Medical; Microarray Analysis; Molecular; Mononuclear; Mus; Oxides; Pathogenesis; Pathologic; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Phase; Phenotype; Phospholipase; Play; Production; Proteins; Publications; Publishing; Pulmonary Hypertension; Rattus; Recombinants; Reporting; Rodent; Role; Small Intestines; Stimulus; Supporting Cell; Symptoms; T-Lymphocyte; TNF gene; TNFSF10 gene; Time; Up-Regulation; Wild Type Mouse; base; cytokine; dietary; experimental study; gut inflammation; improved; intestinal epithelium; knock-down; new therapeutic target; novel; oxidized lipid; peripheral blood; prevent; pulmonary arterial hypertension; pulmonary arterial pressure; right ventricular failure; transcriptome sequencing

SUMMARY Pulmonary arterial hypertension (PAH) is a chronic lung disease characterized by a progressive increase in pulmonary arterial pressure leading to right ventricular (RV) heart failure and death. Over the last five years, our group and others have demonstrated the critical role of oxidized fatty acids in the pathogenesis of PAH. Levels of oxidized fatty acids of the lipoxygenase (LOX) pathway; hydroxyeicosatetraenoic acids (HETEs) are upregulated in the lungs of patients with PAH as well as in rats with pulmonary hypertension (PH). We were the first to establish that dietary supplementation of a single oxidized fatty acid (15HETE) of the LOX pathway is sufficient to cause PH in wild type mice in the absence of any other PH stimulus. In our very recent publication in Hypertension, we reported T cell-dependent endothelial cell apoptosis as one of the mechanisms underlying 15HETE induced PH. However, the exact molecular mechanisms leading to PAEC apoptosis and onset of PH are not known. To unravel the molecular mechanisms, we performed RNA-Seq on the lungs and intestine of mice on 15HETE diet and integrated our RNA-Seq data with online microarrays of human PAH lungs and identified IFI44 (IFN inducible protein 44) as the only novel common gene that was significantly upregulated. IFI44 is an interferon inducible protein and our preliminary data shows that IFNα4 is specifically increased in the small intestine of mice on 15HETE diet. Our preliminary time course experiments revealed increased expression of IFI44 in the small intestine, which precedes its upregulation in the lung, suggesting that 15HETE may act on the small intestine initially. In addition, our pilot data shows that IFI44 is expressed in the immune cells in the lungs of human PAH patients and in T cells in mouse lungs on 15HETE diet. Our bioinformatic analysis also revealed that expression of IFI44 in immune cells in the lungs correlates with CXCL10 (a proinflammatory cytokine) and TRAIL (Tumor Necrosis Factor Related Apoptosis Inducing Ligand). TRAIL and CXCL10 are both known to induce endothelial cell apoptosis. Our preliminary data also shows i) Knockdown of IFI44 in the lungs of mice on 15HETE diet prevented development of PH, and ii) blocking the pathologic action of Cxcl10 rescues PH development. Taken together, our overall hypotheses are i) dietary 15HETE acts on intestinal epithelial cells to produce IFN4, which induces IFI44 in specific immune cells of the Lamina Propria. IFI44 positive immune cells migrate to the lungs and in coordination with CXCL10/TRAIL trigger PAEC death, causing PH; and ii) IFI44 and CXCL10 can serve as novel therapeutic targets in the lungs to prevent or even rescue development of PH in mice on 15HETE diet. Aim 1 will determine the mechanisms by which 15HETE activates the intestinal epithelium resulting in the development of PH; Aim 2 will examine how activation of IFI44 in immune cells in the small intestine induces endothelial cell apoptosis in the lungs through CXC10 and/or TRAIL dependent mechanisms causing pulmonary hypertension; and Aim 3 will examine whether IFI44 and CXCL10 can serve as novel therapeutic targets in the lungs to prevent or rescue development of PH in mice on 15HETE diet.

总结 肺动脉高压（PAH）是一种慢性肺部疾病，其特征是肺动脉高压的进行性增加。 肺动脉压导致右心室（RV）心力衰竭和死亡。在过去的五年里，我们 研究小组和其他研究人员已经证明氧化脂肪酸在PAH发病机制中的关键作用。水平 脂肪氧合酶（LOX）途径的氧化脂肪酸;羟基二十碳四烯酸（HETE）是 在PAH患者和肺动脉高压（PH）大鼠的肺中上调。我们是 首先确定膳食补充LOX途径的单一氧化脂肪酸（15 HETE）是 足以在没有任何其他PH刺激的情况下在野生型小鼠中引起PH。在我们最近出版的 在高血压中，我们报道T细胞依赖性内皮细胞凋亡是高血压的潜在机制之一， 15 HETE可诱导肺动脉高压的发生，但导致肺动脉高压发生的确切分子机制尚不清楚 不知道。为了解开分子机制，我们对肺和肠进行了RNA-Seq， 小鼠15 HETE饮食，并将我们的RNA-Seq数据与人类PAH肺的在线微阵列整合， 鉴定IFI 44（IFN诱导蛋白44）为唯一显著上调的新的共同基因。 IFI 44是一种干扰素诱导蛋白，我们的初步数据表明，IFNα4在人乳腺癌中特异性增加， 15 HETE饮食的小鼠小肠。我们初步的时间进程实验显示， IFI 44在小肠中的表达，这先于其在肺中的上调，表明15 HETE可能作用于 小肠最初。此外，我们的试验数据显示，IFI 44在小鼠的免疫细胞中表达。 人PAH患者肺中的T细胞和15 HETE饮食的小鼠肺中的T细胞。我们的生物信息学分析也 显示肺中免疫细胞中IFI 44的表达与CXCL 10（促炎因子）相关， 细胞因子）和TRAIL（肿瘤坏死因子相关凋亡诱导配体）。TRAIL和CXCL 10都是 已知诱导内皮细胞凋亡。我们的初步数据还显示i）肺中IFI 44的敲低 15 HETE饮食的小鼠的PH预防发展，和ii）阻断Cxcl 10拯救的病理作用 PH发展。综上所述，我们的总体假设是：i）膳食15 HETE作用于肠上皮细胞 以产生IFN β 4，其在固有层的特异性免疫细胞中诱导IFI 44。IFI 44阳性免疫 细胞迁移到肺并与CXCL 10/TRAIL协调触发PAEC死亡，引起PH;和ii）IFI 44 和CXCL 10可以作为肺中新的治疗靶点，以预防甚至挽救PH的发展。 在15 HETE饮食的小鼠中。目的1将确定15 HETE激活肠道的机制， 目的2将研究在PH的发展导致的免疫细胞中IFI 44的激活， 小肠通过CXC 10和/或TRAIL依赖性诱导肺内皮细胞凋亡 目的3将检查IFI 44和CXCL 10是否可以作为肺动脉高压的机制。 肺中的新治疗靶点，以预防或挽救15 HETE饮食小鼠中PH的发展。