Chemogenomic Profiling of Plasmodium Falciparum Responses and Resistance

项目摘要/摘要 疟疾是人类死亡和疾病的主要原因，造成超过2亿的临床病例。 疟疾和每年40万人死亡。控制和治愈疟疾的传统措施是 受到青蒿素耐药性出现的威胁(ART-R)。对ART-R的研究集中在 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 是由ART的作用机制造成的。然而，最近的发现表明，抗药性- K13基因的相关突变会减缓细胞胞体的功能，从而减少可利用的血红蛋白 食物液泡。我们的初步结果显示，寄生虫对 ART与使恶性疟原虫存活的先天应激反应通路显著重叠 疟疾热。我们的实验方法是阐明药物与基因的关联并破译 利用正向遗传技术研究抗逆转录病毒药物和其他抗疟疾药物的作用机制和耐药性 随机猪Bac诱变恶性疟原虫突变株的筛选。这种方法具有 确定了对恶性疟原虫至关重要的主要寄生虫过程中的基因突变 发热生存反应与ART(DHA，AS)敏感性的改变显著相关，表明 这种寄生虫劫持了热休克应激反应途径，以应对ART毒性。我们会 使用小的iggyBac克隆库和专注于Go的库来迭代筛选不同的 从功能上注释交互伙伴、途径和监管流程的表型 与ART的作用机制和抵抗力有关。我们将使用基因组水平的筛选来识别 与抗逆转录病毒治疗作用机制相关的因素。我们将把我们的分析扩展到P.nowlesi 通过适应和应用来表征保守的高价值抗疟疾药物靶点 这种间日疟原虫的化学基因组图谱分析。

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.