tRNA modification reprogramming in artemisinin-resistant Plasmodium falciparum: An epigenetic driver of resistance?

抗青蒿素恶性疟原虫中的 tRNA 修饰重编程：耐药性的表观遗传驱动因素？

• 批准号: 10404060
• 负责人: Jennifer L. Small-Saunders
• 金额: $ 19.22万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10404060
• 关键词: Acetylation; Acetyltransferase; Africa; Amino Acyl Transfer RNA; Aminoacyl-tRNA hydrolase; Artemisinins; Asia; Biology; Biomedical Research; Blood; Cellular Stress; Codon Nucleotides; Collection; Communicable Diseases; Complex; Coupled; Data; Development; Dimethyl Sulfoxide; Doctor of Philosophy; Drug resistance; Endocytosis; Engineering; Environment; Enzymes; Epigenetic Process; Eukaryota; Exposure to; Genes; Genetic; Genetic Transcription; Hemoglobin; Immunology; Kinetics; Laboratories; Lead; Liquid Chromatography; Malaria; Mass Spectrum Analysis; Mediating; Medical center; Medicine; Mentors; Metabolic; Microbiology; Modification; Molecular; Mutation; Nutritional; Parasite resistance; Parasites; Parasitology; Pathway interactions; Patient Care; Pharmaceutical Preparations; Physicians; Physiologic pulse; Plasmodium falciparum; Play; Positioning Attribute; Process; Prokaryotic Cells; Proteins; Proteome; Proteomics; RNA; Reaction Time; Reproducibility; Research; Resistance; Resistance development; Ribosomes; Role; Sampling; Scientist; Secondary to; South America; Stress; System; Techniques; Testing; Time; Training; Transcriptional Regulation; Transfer RNA; Translational Regulation; Translations; Universities; Up-Regulation; Variant; asexual; base; biological adaptation to stress; career; environmental enrichment for laboratory animals; epigenetic regulation; experimental study; genetic approach; innovation; insight; knock-down; multidisciplinary; multiple omics; mutant; pressure; resistant Plasmodium falciparum; response; sensor; tandem mass spectrometry; tool; transcriptome sequencing; translational proteomics

PROJECT SUMMARY / ABSTRACT: Rationale: The spread of artemisinin (ART)-resistant Plasmodium falciparum (Pf) strains across Asia and their recent emergence in Africa and South America imperils efforts to treat and control malaria. ART resistance is mediated primarily by mutations in Pf K13, which reduce drug activation by decreasing hemoglobin endocytosis and which initiate quiescence during peak drug levels. Our preliminary data have uncovered a role for tRNA modification reprogramming and proteomic changes in mutant K13 parasites exposed to ART pressure. In this mentored career project, we hypothesize that quiescence is epigenetically regulated by reprogramming tRNA modifications, which leverage codon-biased translation to alter the parasite proteome and enable the survival of ART-treated mutant K13 parasites. Candidate: As an Infectious Diseases physician with a PhD in Microbiology and Immunology, I am uniquely positioned to bridge biomedical research and patient care to understand the molecular mechanisms that Pf employs to survive ART treatment. Further training in molecular parasitology, mass spectrometry, proteomics, RNA biology, and epigenetics will be crucial for my development into an independent academic physician-scientist specializing in Pf stress responses and drug resistance. I have a renowned mentor in Dr. David Fidock and benefit from an outstanding multi- disciplinary team of experts to guide my training and research progress. Environment: The Fidock laboratory at the Columbia University Irving Medical Center (CUIMC) has been a pioneer in applying genetic and multi-omic tools to explore Pf resistance to ART and other drugs. This enriching environment also provides access to multiple isogenic k13-edited Pf lines and a large network of collaborators including experts in mass spectrometry- based tRNA modifications, proteomics, and codon-biased translation. CUIMC also has a long track record of enabling young physician-scientists to develop independent and successful careers in academic medicine. Approach: Our central hypothesis is that tRNA modification reprogramming, specifically the s2U modification, is central to how parasites achieve ART resistance by altering their proteome and regulating entry into and exit from drug-induced quiescence. In Aim 1, we will elucidate the kinetics of tRNA modification reprogramming in isogenic ART-resistant and ART-sensitive parasites across a panel of K13 variants and genetic backgrounds. In Aim 2, we will apply conditional knockdown approaches to explore the role of the s2U pathway in ART resistance and parasite survival. In Aim 3, we will test the complementary hypothesis that modifications on the amino-acyl tRNA regulate ART-mediated quiescence. This proposal provides an innovative approach to examining how K13 mutations achieve ART resistance via epigenetic changes that reprogram tRNA modifications to alter translational and proteomic responses to ART pressure. If confirmed experimentally, these hypotheses will set a new paradigm for how Pf can modulate its biology to survive drug-mediated cellular stress. Our results should also identify parasite vulnerabilities that can be leveraged into new strategies to treat ART-resistant malaria.

项目摘要 /摘要：基本原理：阿秒蛋白（ART）抗性疟原虫的传播 遍布亚洲的恶性菌（PF）菌株及其最近在非洲和南美的出现危及了努力 治疗和控制疟疾。 ART抗性主要是由PF K13中的突变介导的，从而减少药物 通过降低血红蛋白内吞作用并在峰值药物水平期间引发静止来激活。我们的 初步数据已经发现了tRNA修饰重编程和突变体蛋白质组学变化的作用 K13寄生虫受到艺术压力。在这个指导的职业项目中，我们假设静止是 通过重编程tRNA修改对表观遗传学调节，该修饰利用密码子偏见的翻译来改变 寄生虫蛋白质组，并使艺术处理的突变体K13寄生虫的生存。候选人：作为传染性 具有微生物学和免疫学博士学位的疾病医师，我在桥梁生物医学方面拥有独特的位置 研究和患者护理了解PF在艺术治疗中使用的分子机制。 在分子寄生虫学，质谱，蛋白质组学，RNA生物学和表观遗传学方面的进一步培训将是 对于我发展为独立的学术医师科学家，专门从事PF压力反应的至关重要 和耐药性。我在戴维·菲达克（David Fidock）博士中有一位著名的导师 纪律专家团队指导我的培训和研究进度。环境：Fidock实验室 哥伦比亚大学欧文医学中心（CUIMC）一直是应用遗传和多运动的先驱 探索PF抵抗艺术和其他药物的工具。这种丰富的环境还提供了访问 多种同源K13编辑的PF线和大型合作者网络，包括质谱专家 基于基于的tRNA修饰，蛋白质组学和密码子偏见的翻译。 Cuimc也有很长的记录 使年轻的医师科学家能够发展在学术医学领域的独立和成功职业。 方法：我们的中心假设是tRNA修改重编程，特别是S2U修饰，是 寄生虫如何通过改变其蛋白质组并调节进入和退出的核心 来自药物诱导的静止。在AIM 1中，我们将阐明tRNA修饰的动力学重新编程 一组K13变体和遗传背景的耐同源艺术和艺术敏感的寄生虫。在 AIM 2，我们将采用有条件的敲低方法来探索S2U途径在艺术阻力中的作用 和寄生虫生存。在AIM 3中，我们将测试互补假设，即对氨基酰基的修改 tRNA调节艺术介导的静止。该建议提供了一种创新的方法来研究K13 突变通过表观遗传学变化实现ART抗性，重新编程tRNA修饰以改变 翻译和蛋白质组学对艺术压力的反应。如果通过实验确认，这些假设将设定 PF如何调节其生物学以生存药物介导的细胞应激的新范式。我们的结果应该 还要确定可以利用的寄生虫脆弱性，这些脆弱性可以利用新的策略来治疗抗艺术疟疾。