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

项目总结/摘要 血吸虫病引起的肺动脉高压（Sch-PH）是全球最常见的肺动脉高压病因。 肺动脉高压（PAH），其分类为"被忽视的热带疾病"强调了 在理解疾病病理生物学和开发有效治疗方法方面存在大量未满足的需求。虽然 越来越多的数据支持转录因子缺氧诱导因子-1（HIF-1）的可能作用， HIF-1调节糖酵解对PH发病机制的影响，其在Sch-PH中的意义尚不清楚。这 一项提案研究了HIF-1依赖性糖酵解在促进Sch-PH中的潜在作用，特别是在肺中 间质巨噬细胞（IM）。现有数据表明，肺血管周围IM浸润增加 在患有Sch-PH的人类和实验小鼠中，HIF-1相关基因的转录增加， Sch-PH小鼠肺IM中的糖酵解酶;以及LsyM+髓样细胞中HIF-1 α缺失的保护作用 基于这些关键的见解，这项建议将测试假设，HIF-1- 血管周围肺IM中的依赖性糖酵解对Sch-PH的发展起着关键作用。 糖酵解-乳酸发酵，磷酸戊糖途径， 和线粒体Krebs循环-乳酸和肺动脉平滑之间的潜在机制联系 将另外检查肌细胞（PASMC）增殖。提出了两个目标。目标1将决定 通过条件性缺失和稳定HIF-1 α来稳定Sch-PH中肺IM HIF-1 α的必要性和充分性 1 α在转基因小鼠IM中的表达。乳酸脱氢酶A（LDH-A;一种负责乳酸的酶） 将检查Sch-PH严重性的缺失和草氨酸盐（LDH-A抑制剂）处理。目标2将 时空表型肺IM，量化其葡萄糖代谢，并使用共培养测试肺IM- 衍生的乳酸盐在PASMC中诱导促增殖、促纤维化表型。建成后将 阐明肺IM中的葡萄糖代谢如何有助于Sch-PH病理生物学。 拟议的职业发展计划的最终目标是支持申请人的 在肺血管代谢领域成功转型为独立的临床医生-科学家。的 计划利用他的导师和顾问在基础翻译研究，他们的 对导师的承诺，以及UCSF研究和专业发展的集体资源。在 通过五年的培训，申请人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;海马代谢测定;质谱;和共聚焦 显微镜），发展数据分析技能，传播他的发现，并共同完善花岗岩 将他定位为肺血管代谢领域的专家调查员。

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.