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

肺间质巨噬细胞缺氧诱导因子1依赖性糖酵解对血吸虫病引起的肺动脉高压病理学的贡献。

• 批准号: 10644936
• 负责人: Michael Hyunjean Lee
• 金额: $ 16.9万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-05 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10644936
• 关键词: Address; Biological Assay; Blood Vessels; Carbon; Cell Proliferation; Cell Separation; Cessation of life; Citric Acid Cycle; Classification; Coculture Techniques; Confocal Microscopy; Data; Data Analytics; Development; Development Plans; Disease; Enzymes; Fermentation; Flow Cytometry; Genes; Genetic Transcription; Genus Hippocampus; Glucose; Glycolysis; Glycolysis Induction; Goals; Grant; Growth; High Prevalence; Human; In Situ; Infiltration; Knowledge; Label; Lesion; Link; Lung; Macrophage; Mass Spectrum Analysis; Medial; Mentors; Mentorship; Metabolic; Metabolism; Mitochondria; Mus; Myeloid Cells; Oral; Pathogenesis; Pathologic; Pentosephosphate Pathway; Phenocopy; Phenotype; Positioning Attribute; Production; Proliferating; Proteomics; Pulmonary Hypertension; Pulmonary arterial remodeling; Pulmonary artery structure; Research Personnel; Resistance; Resources; Role; Schistosomiasis; Scientist; Severities; Smooth Muscle Myocytes; Structure of parenchyma of lung; Tamoxifen; Testing; Therapeutic; Thick; Time; Training; Transgenic Mice; Translational Research; Up-Regulation; Vascular Proliferation; arterial lesion; career development; design; effective therapy; experimental study; fibrogenesis; glucose metabolism; glucose uptake; hypoxia inducible factor 1; hypoxia-induced pulmonary hypertension; inhibitor; insight; interstitial; lactate dehydrogenase A; neglected tropical diseases; novel; protective effect; protein expression; pulmonary arterial hypertension; pulmonary vascular remodeling; right ventricular failure; skills; spatiotemporal; transcription factor; uptake

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.

项目概要/摘要 血吸虫病引起的肺动脉高压（Sch-PH）是全球最常见的原因 肺动脉高压（PAH）及其被归类为“被忽视的热带疾病”强调了 了解疾病病理学和开发有效治疗方法的巨大需求未得到满足。 积累的数据支持转录因子缺氧诱导因子-1 (HIF-1) 的可能贡献以及 HIF-1 调节糖酵解对 PH 发病机制的影响，其在 Sch-PH 中的意义很大程度上未知。 该提案研究了 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 现有数据表明，间质巨噬细胞（IM）的肺血管周围浸润增加。 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； 具有 Sch-PH 的小鼠肺 IM 中的糖酵解酶以及 LsyM+ 骨髓中 HIF-1α 缺失的保护作用； 基于这些关键见解，该提案将检验 HIF-1- 的假设。 血管周围肺 IM 中的依赖性糖酵解对 Sch-PH 的发展至关重要。 糖酵解的潜在生长促进机制——乳酸发酵、磷酸戊糖途径、 和线粒体克雷布斯循环——乳酸和肺动脉平滑肌之间的潜在机制联系 将进一步检查肌肉细胞 (PASMC) 增殖，并确定目标 1。 通过有条件删除和稳定 HIF- 来稳定 Sch-PH 中肺 IM HIF-1α 的必要性和充分性 转基因小鼠 IM 中的 1α 乳酸脱氢酶 A (LDH-A；一种负责乳酸的酶) 的作用。 将检查删除）删除和草酸盐发酵（LDH-A 抑制剂）处理对 Sch-PH 严重程度的影响。 时空表型肺 IM，量化其葡萄糖代谢，并使用共培养测试肺 IM 是否 衍生的乳酸会在 PASMC 中诱导促增殖、促纤维化表型。 阐明肺 IM 中的葡萄糖代谢如何影响 Sch-PH 病理学。 拟议的职业发展计划旨在支持申请人的最终目标 在肺血管代谢领域成功过渡为独立的临床医生科学家。 该计划利用了他的导师和顾问在基础转化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研究和专业发展的集体资源。 通过五年的培训，申请人将熟练掌握关键实验方法（空间分辨 蛋白质组学和细胞分选； 显微镜），发展数据分析技能，传播他的发现，并完善资助政策，共同 使他成为肺血管代谢领域的专家研究员。