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: 10317747
• 负责人: John H Adams
• 金额: $ 73.64万
• 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/10317747
• 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/antagonist; 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.

项目摘要/摘要 疟疾是人类死亡和疾病的主要原因，导致超过2亿例临床病例 疟疾和每年40万人死亡。控制和治愈疟疾的传统措施是 受青蒿素抵抗（ART-R）的出现威胁。研究ART-R的研究集中 主要是基于允许寄生虫耐受氧化应激和蛋白质损伤的机制 由艺术的作用机理产生。但是，最近的发现表明抗性 - K13中的相关突变减慢了细胞抑制仪的功能，以减少可用的血红蛋白 食品真空吸尘器。我们的初步结果表明，寄生虫对 艺术与先天应力反应途径重叠，使恶性疟原虫生存 疟疾热。我们的实验方法是阐明药物 - 基因的关联和破译 使用正向遗传 由随机piggybac诱变产生的恶性疟原虫突变体的筛选。这种方法有 确定主要寄生虫过程中的遗传突变对于恶性疟原虫疟疾至关重要 发烧生存反应与对艺术的敏感性的改变显着相关，表明 寄生虫劫持了应对艺术毒性的热冲压反应途径。我们将 使用小型盖贝克克隆的小图书馆和以不同的迭代屏幕的方式 表型在功能上注释相互作用的伙伴，途径和调节过程 与动作和抵抗的艺术机理有关。我们将使用基因组级屏幕来识别 与艺术作用机理有关的因素。我们将把分析扩展到P. Knowlesi到 通过适应和应用来表征配置的高价值抗疟药靶标 化学生成分析分析对这种类体类似疟疾的寄生虫。