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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要疟疾是人类死亡和疾病的主要原因，导致超过 2 亿例临床病例疟疾每年导致 40 万人死亡。受到青蒿素耐药性（ART-R）出现的威胁，对 ART-R 的研究已成为重点。主要是关于允许寄生虫耐受氧化应激和蛋白质损伤的机制然而，最近的发现表明，抗性- K13 的相关突变会减慢细胞口功能，从而减少体内可用的血红蛋白我们的初步结果显示寄生虫对食物液泡的敏感性和耐受性。 ART 与使恶性疟原虫得以存活的先天应激反应途径显着重叠我们的实验方法是阐明药物基因关联并破译。使用正向遗传学研究 ART 和其他抗疟药物的作用机制和耐药性通过随机piggyBac诱变产生的恶性疟原虫突变体的筛选。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烧生存反应与 ART（DHA、AS）敏感性改变显着相关，表明寄生虫劫持了热休克应激反应途径来应对 ART 毒性。使用piggyBac克隆的小型文库和以GO为中心的文库来迭代筛选不同的表型对相互作用的伙伴、途径和调控过程进行功能注释与 ART 的作用机制和耐药性相关联，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诺氏假单胞菌通过调整和应用来表征保守的高价值抗疟药物靶点对这种间日疟原虫的化学基因组分析。