The contribution of hypoxia inducible factor-1-dependent glycolysis in lung interstitial macrophages to the pathobiology of schistosomiasis-induced pulmonary hypertension.

项目摘要/摘要 血吸虫病诱发的肺动脉高压(SCH-PH)是全球最常见的 肺动脉高压(PAH)及其被归类为“被忽视的热带疾病”强调 在了解疾病的病理生物学和开发有效的治疗方法方面存在大量未得到满足的需求。虽然 越来越多的数据支持转录因子缺氧诱导因子-1(HIF-1)和 HIF-1调节糖酵解在PH发病机制中的作用，其在Sch-PH中的意义尚不清楚。这 一项提案研究了依赖于HIF-1的糖酵解在促进Sch-PH中的潜在作用，特别是在肺中 间质巨噬细胞(IMS)。现有数据显示IMS肺血管周围浸润增加 在人类和实验性Sch-PH小鼠中；HIF-1相关基因编码的转录增加 系统性红斑狼疮小鼠肺间质干细胞糖酵解酶及髓系细胞缺氧诱导因子-1α缺失的保护作用 细胞抗小鼠Sch-PH。在这些关键见解的基础上，这项提议将检验HIF-1-1- 血管周围肺IMS的依赖性糖酵解在SCH-PH的发生发展中起关键作用。三个人中的一个 糖酵解潜在的促生长机制--乳酸发酵，磷酸戊糖途径， 线粒体Krebs循环--乳酸和肺动脉通畅之间潜在的机制联系 另外还将检查肌肉细胞(PASMC)的增殖情况。提出了两个目标。目标一号将决定 选择性删除和稳定肺组织IM-HIF-1α的必要性和充分性 1转基因小鼠免疫球蛋白中的α。乳酸脱氢酶A(LDH-A)的作用 发酵)缺失和草酸酯(LDH-A抑制剂)处理对SCH-PH严重性的影响将被考察。目标2将 肺IM的时空表型，量化它们的葡萄糖代谢，并使用共培养法检测是否肺IM- 衍生乳酸盐在PASMCs中诱导促增殖、促纤维化表型。该项目的完成将 阐明肺IMS中的葡萄糖代谢如何在SCH-PH病理生物学中起作用。 拟议的职业发展计划的最终目标是支持申请者的 作为一名临床医生兼科学家，在肺血管代谢领域成功地过渡到独立。这个 Plans利用了他的导师和顾问在基础翻译研究方面的综合专业知识，他们的 致力于指导，以及加州大学旧金山分校用于研究和专业发展的集体资源。在……里面 在五年的培训中，申请者将熟练掌握关键的实验方法(空间分辨 蛋白质组学；流式细胞术和细胞分选；海马代谢分析；质谱学；以及共聚焦 显微镜)，发展数据分析技能，传播他的发现，并共同完善格兰特技巧 将他定位为肺血管新陈代谢领域的专家研究员。

Project Summary/Abstract Schistosomiasis-induced pulmonary hypertension (Sch-PH) is globally the most common cause of pulmonary arterial hypertension (PAH), and its classification as a “neglected tropical disease” underscores the large unmet need in understanding the disease pathobiology and developing effective treatments. Although accumulating data support the likely contribution of the transcription factor hypoxia inducible factor-1 (HIF-1) and HIF-1-regulated glycolysis to the pathogenesis of PH, their significance in Sch-PH is largely unknown. This proposal examines the potential role of HIF-1-dependent glycolysis in promoting Sch-PH, specifically in lung interstitial macrophages (IMs). Available data demonstrate increased pulmonary perivascular infiltration of IMs in both humans and experimental mice with Sch-PH; increased transcription of HIF-1-associated genes encoding glycolytic enzymes in murine lung IMs with Sch-PH; and a protective effect of HIF-1α deletion in LsyM+ myeloid cells against murine Sch-PH. Building upon these key insights, this proposal will test the hypothesis that HIF-1- dependent glycolysis in perivascular lung IMs critically contributes to the development of Sch-PH. Of the three potential growth-promoting mechanisms of glycolysis--lactate fermentation, the pentose phosphate pathway, and the mitochondrial Krebs cycle--the potential mechanistic link between lactate and pulmonary artery smooth muscle cell (PASMC) proliferation will be additionally examined. Two aims are proposed. Aim 1 will determine necessity and sufficiency of lung IM HIF-1α stabilization in Sch-PH by conditionally deleting and stabilizing HIF- 1α in IMs of transgenic mice. The effect of lactate dehydrogenase A (LDH-A; an enzyme responsible for lactate fermentation) deletion and oxamate (LDH-A inhibitor) treatment on Sch-PH severity will be examined. Aim 2 will spatiotemporally phenotype lung IMs, quantify their glucose metabolism, and test, using co-culture, if lung IM- derived lactate induces a pro-proliferative, pro-fibrogenic phenotype in PASMCs. Completion of the project will clarify how glucose metabolism in lung IMs contributes to Sch-PH pathobiology. The proposed career development plan was designed with the ultimate goal of supporting the applicant’s successful transition to independence as a clinician-scientist, in the field of pulmonary vascular metabolism. The plan leverages the combined expertise of his mentors and advisors in basic-translational research, their commitment to mentorship, and the collective resources for research and professional development at UCSF. In the five years of training, the applicant will acquire proficiency in key experimental approaches (spatially resolved proteomics; flow cytometry and cell sorting; Seahorse metabolic assay; mass spectrometry; and confocal microscopy), develop data analytical skills, disseminate his findings, and refine grantsmanship, collectively positioning him as an expert investigator in the field of pulmonary vascular metabolism.