Chemogenomic Profiling of Plasmodium Falciparum Responses and Resistance

恶性疟原虫反应和耐药性的化学基因组学分析

• 批准号: 10449354
• 负责人: John H Adams
• 金额: $ 69.48万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-10 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10449354
• 关键词: Africa; Anabolism; Anti-malarial drug resistance; Antimalarials; Artemisinins; Asia; Blood; Blood Circulation; CRISPR/Cas technology; Cessation of life; Clinical; Combined Modality Therapy; Complex; Coupled; DNA Sequence Alteration; Development; Digestion; Disease; Drug Targeting; Drug usage; Falciparum Malaria; Fever; Food; Gene Expression Profile; Genes; Genetic Crosses; Genetic Screening; Genome; Genomic Library; Genotype; Heat shock proteins; Heat-Shock Response; Heme; Hemoglobin; Human; Libraries; Link; Malaria; Measures; Mediating; Metabolic; Modeling; Mutagenesis; Mutation; Ontology; Oxidative Stress; Parasites; Pathway interactions; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Plasmodium; Plasmodium falciparum; Plasmodium vivax; Process; Proteins; Reactive Oxygen Species; Research; Resistance; Toxic effect; Vacuole; artesunate; asexual; biological adaptation to stress; comparative; cost; docosahexaenoylascorbic acid; drug discovery; fitness; gene discovery; genome editing; global health; inhibitor; isoprenoid; multicatalytic endopeptidase complex; mutant; response; tool; transcriptome sequencing; whole genome

Project Summary/Abstract Malaria is a leading cause of human death and illness, causing over 200 million cases of clinical malaria and 400,000 deaths each year. Traditional measures to control and cure malaria are threatened by emergence of artemisinin resistance (ART-R). Research into ART-R has focused mostly on mechanisms allowing parasite to tolerate the oxidative stress and protein damage resulting from ART’s mechanism of action. However, recent discoveries indicate that resistance- associated mutations in the K13 slows cytostome function to diminish the available hemoglobin in the food vacuole. Our preliminary results revealed that the parasite’s sensitivity and tolerance to ART significantly overlaps with innate stress response pathways that enable P. falciparum survival of malaria fever. Our experimental approach is to elucidate drug-gene associations and decipher mechanisms of action and resistance to ART and other antimalarial drugs, using forward genetic screens of P. falciparum mutants created by random piggyBac mutagenesis. This approach has determined that genetic mutations in the major parasite processes critical for P. falciparum malarial fever survival response significantly correlate with altered sensitivity to ART (DHA, AS), indicating the parasite hijacked the heat-shock stress response pathways to cope with ART toxicity. We will use small libraries of piggyBac clones and GO-focused libraries for iterative screens of different phenotypes to functionally annotate interacting partners, pathways, and regulatory processes linked to ART mechanism of action and resistance. We will use genome-level screens to identify factors linked to ART mechanism of action. We will extend our analysis to P. knowlesi to characterize the conserved high-value antimalarial drug targets by adapting and applying chemogenomic profiling analysis to this vivax-like malaria parasite.

项目总结/摘要 疟疾是人类死亡和疾病的主要原因，造成超过2亿例临床病例， 每年有40万人死于疟疾。控制和治疗疟疾的传统措施是 受到青蒿素耐药性（ART-R）出现的威胁。对ART-R的研究集中在 主要研究寄生虫耐受氧化应激和蛋白质损伤的机制 这是由于ART的作用机制。然而，最近的发现表明，耐药性- K13的相关突变减缓了细胞口功能，从而减少了细胞中可用的血红蛋白。 食物泡我们的初步结果表明，寄生虫的敏感性和耐受性 ART与使恶性疟原虫存活的先天应激反应途径显著重叠 疟疾发烧。我们的实验方法是阐明药物基因协会和破译 使用正向遗传学方法研究抗逆转录病毒疗法和其他抗疟药物的作用机制和耐药性 筛选通过随机piggyBac诱变产生的恶性疟原虫突变体。这种方法有 确定主要寄生虫过程中的基因突变对恶性疟原虫疟疾至关重要， 发热存活反应与ART敏感性改变（DHA，AS）显著相关，表明 寄生虫劫持了热休克应激反应途径来科普ART毒性。我们将 使用piggyBac克隆的小文库和GO聚焦文库进行不同的重复筛选， 表型功能注释相互作用的伙伴，途径和监管过程 与抗逆转录病毒疗法的作用机制和耐药性有关。我们将使用基因组水平的筛选来识别 与ART作用机制相关的因素。我们将把我们的分析扩展到P. knowlesi， 通过调整和应用， 化学基因组分析，这种间日疟原虫样疟疾寄生虫。