Role of host fatty acid metabolism in Trypanosoma cruzi amastigote growth

宿主脂肪酸代谢在克氏锥虫无鞭毛体生长中的作用

• 批准号: 9056973
• 负责人: BARBARA A BURLEIGH
• 金额: $ 40.38万
• 依托单位: HARVARD SCHOOL OF PUBLIC HEALTH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9056973
• 关键词: Address; Albumins; Amino Acids; Binding; Biochemical; Biological; Biology; CD36 gene; Carbohydrates; Cardiac Myocytes; Catabolic Process; Cell surface; Cells; Chagas Disease; Chronic; Collaborations; Critical Pathways; Data; Dependency; Development; Disease Outcome; Disease Progression; Energy-Generating Resources; Enzymes; Exhibits; FABP3 gene; Fatty Acids; Gene Targeting; Generations; Genes; Goals; Growth; Health; Hela Cells; Human; Hydrolysis; Immigrant; Infection; Intercept; Knowledge; Laboratories; Latin America; Lipids; Mammalian Cell; Mass Spectrum Analysis; Measures; Mediating; Membrane Lipids; Metabolic; Metabolic Pathway; Mitochondria; Modeling; Molecular; Mus; Myocardial; Myocardium; Nonesterified Fatty Acids; Nutrient; Parasites; Parasitic Diseases; Pathogenesis; Pathway interactions; Patients; Polyamines; Positioning Attribute; Predisposition; Process; Purines; RNA Interference; RNA interference screen; Role; Smooth Muscle; System; Therapeutic; Therapeutic Intervention; Triglycerides; Trypanosoma cruzi; Very Long Chain Fatty Acid; Work; base; cell type; chagasic cardiomyopathy; design; extracellular; fatty acid elongases; fatty acid metabolism; fatty acid transport; flexibility; genome-wide; improved; induced pluripotent stem cell; insight; knock-down; knockout gene; long chain fatty acid; membrane synthesis; metabolic phenotype; metabolomics; mutant; novel; novel strategies; obligate intracellular parasite; oxidation; pathogen; research study; small molecule; tool; uptake

 DESCRIPTION (provided by applicant): The Chagas' disease parasite Trypanosoma cruzi has the ability to infect many nucleated cell types, but its persistence in cardiac and smooth muscle is critical for disease progression. Like all obligate intracellular pathogens, T. cruzi mus couple its metabolic requirements to its host in order to survive. If the biochemical and cellular pathways that tether T. cruzi to its host cell were known - for cardiomyocytes in particular - we would be in a stronger position to design strategies to disrupt these functional ties and to inhibi parasite infection. However, with the exception of the well-recognized auxotrophies for purines, polyamines and certain amino acids, the metabolic dependencies of intracellular T. cruzi amastigotes have not been described in any cell type. This proposal builds on results from a recent genome-wide RNA interference (RNAi) screen in HeLa cells that offers an unbiased preview of host susceptibility factors that support intracellular T. cruzi infection. Among the pathways identified in our screen, host fatty acid metabolism emerged as a prominent candidate pathway that positively impacts intracellular T. cruzi amastigote growth. Proposed studies seek to determine where and how intracellular T. cruzi amastigotes intersect host fatty acid ß-oxidation and synthesis pathways and where the critical dependencies for this pathogen lie. Studies will exploit induced pluripotent stem cell-derived (iPSC) human cardiomyocytes as a new and biologically relevant T. cruzi infection model. RNAi will be combined with extracellular metabolic flux analyses and metabolic mass spectrometry to identify specific components (lipid entities and host enzymes) of host fatty acid metabolism that are critical to supporting human cardiomyocyte infection by T. cruzi. The proposed molecular and phenotypic characterization of host fatty acid metabolism as a key regulator of intracellular T. cruzi growth in cardiomyocytes will generate novel insights into the biology of this important human pathogen and its ability to integrate its metabolic needs with the host. Knowledge of these core dependencies has the potential to inform therapeutic strategies aimed at uncoupling the pathogen from its host.

 描述（由申请人提供）：查加斯病寄生虫克氏锥虫能够感染多种有核细胞类型，但其在心脏和平滑肌中的持续存在对疾病进展至关重要。像所有的专性细胞内病原体，T。cruzi mus将其代谢需求与其宿主结合以生存。如果将T. cruzi对宿主细胞的作用是已知的--特别是对心肌细胞--我们将处于更有利的地位来设计策略来破坏这些功能联系和抗寄生虫感染。然而，除了公认的嘌呤、多胺和某些氨基酸营养缺陷型外，细胞内T。尚未在任何细胞类型中描述过克鲁齐无鞭体。该建议建立在最近在HeLa细胞中进行的全基因组RNA干扰（RNAi）筛选的结果上，该筛选提供了支持细胞内T细胞的宿主易感性因子的无偏见预览。克氏感染在我们筛选的途径中，宿主脂肪酸代谢作为一个突出的候选途径出现，对细胞内T细胞产生积极影响。cruzi无鞭毛体生长。拟议的研究试图确定细胞内T。克氏无鞭毛体与宿主脂肪酸β-氧化和合成途径相交，并且是该病原体的关键依赖性所在。研究将利用诱导多能干细胞衍生的（iPSC）人心肌细胞作为一种新的和生物学相关的T。Cruzi感染模型RNAi将与细胞外代谢通量分析和代谢质谱法相结合，以鉴定宿主脂肪酸代谢的特定组分（脂质实体和宿主酶），这些组分对支持T.克鲁兹宿主脂肪酸代谢作为细胞内T。cruzi在心肌细胞中的生长将产生对这种重要的人类病原体的生物学及其将其代谢需求与宿主整合的能力的新见解。这些核心依赖性的知识有可能为旨在将病原体与其宿主解偶联的治疗策略提供信息。