Discovering resistance-resistant antimalarial drug target

项目摘要/摘要 疟疾仍然是全球卫生的主要负担，感染人数超过2.2亿，死亡人数超过400人。 每年有几千人。尽管过去15年来加强国际努力消除 疟疾在全球范围内降低了疟疾发病率，但这一进展在最近几年停滞不前。 抗疟疾药物是减少疾病和传播的基础，但已经产生了抗药性 自1945年以来开发的每一种抗疟疾药物在临床引入12年内，包括 目前以青蒿素为基础的一线联合疗法。寄生虫发展的这种简易性 耐药性清楚地表明，抗疟疾药物发现的创新方法迫在眉睫 需要的。在这里，我们提出了两种不同的策略，旨在发现抗疟疾药物的靶点 哪些抗性将很难在田间形成。第一种方法将描述新陈代谢 疟疾寄生虫对原药产生抗药性时获得的适应，重点是 识别耐药寄生虫的新陈代谢依赖(脆弱性)。我们建议聘用 我们在疟疾寄生虫、疟原虫代谢组学特征方面的丰富经验 恶性疟原虫，比较药物敏感和耐药寄生虫的生化结构。 代谢组学将被用来阐明耐药寄生虫中改变的代谢途径，以便 揭示侧枝敏感性，我们将使用这两个小分子中的任何一个作为假定的药物靶点进行验证 途径抑制物或相关靶标酶的基因敲除。最终的目标将是瞄准 这些侧支敏感性与主要药物一起将寄生虫锁定为药物敏感 状态，因为初级电阻所需的适应不再可用，因此提供了 持久、互补的联合疗法。对于第二种策略，我们建议发现主机 支持恶性疟原虫血期发育的关键代谢过程。鲜血 疟疾寄生虫生命周期的各个阶段都是导致症状和人与人之间传播的原因。 蚊子传播寄生虫，严重依赖人类血清中的营养物质。然而，我们 已经发现不同的人血清在支持寄生虫生长的能力上有很大的差异 和变速箱。为了确定与寄生虫表型读数相关的血清代谢物，我们 建议将我们的新陈代谢方法与标准寄生虫生长和 传播化验。使用健康捐献者的大量血清，这种方法将自然识别 寄生虫所依赖的可变的、因此潜在的可定向的宿主进程。瞄准 寄主因素使寄生虫失去了进化的控制力，降低了抗药性的可能性。 浮现。总而言之，这项提案旨在解决长期存在的抗疟疾药物耐药性问题 通过评估发现耐药药物靶点的两种新战略来发展。

PROJECT SUMMARY/ABSTRACT Malaria continues to be a major burden on global health, infecting over 220 million and killing over 400 thousand people annually. Although the past fifteen years of enhanced international efforts to eradicate malaria has reduced malaria incidence worldwide, this progress has stagnated in recent years. Antimalarial drugs are foundational in reducing disease and transmission, but resistance has developed within twelve years of clinical introduction to every antimalarial drug developed since 1945 including the current front-line artemisinin-based combination therapies. This ease with which parasites develop resistance clearly indicates that innovative approaches to antimalarial drug discovery are urgently needed. Here we propose two separate strategies designed to discover antimalarial drug targets for which resistance will be difficult to develop in the field. The first approach will characterize metabolic adaptations acquired by malaria parasites upon becoming resistant to a primary drug, with a focus on identifying novel metabolic dependencies (vulnerabilities) of resistant parasites. We propose to employ our extensive experience in metabolomic characterization of the malaria parasite, Plasmodium falciparum, to compare drug-sensitive and drug-resistant parasite biochemical architectures. Metabolomics will be used to elucidate altered metabolic pathways in resistant parasites in order to reveal collateral sensitivities that we will validate as putative drug targets using either small molecule pathway inhibitors or genetic knockdowns of relevant target enzymes. The ultimate goal will be to target these collateral sensitivities together with the primary drug to lock the parasite into a drug-sensitive state, as the adaptations required for primary resistance are no longer available, thus providing an enduring, complementary combination therapy. For the second strategy, we propose to discover host metabolic processes that are critical to support blood-stage development of P. falciparum. The blood stages of the malaria parasite life cycle are responsible for both the symptomology and the human-to- mosquito transmission of the parasite, and critically rely on nutrients from human serum. However, we have found that different human serum lots are highly variable in their ability to support parasite growth and transmission. To identify serum metabolites that correlate with parasite phenotypic readouts, we propose to utilize our metabolomic approaches in combination with standard parasite growth and transmission assays. Using serum lots from healthy donors, this approach will identify naturally variable, and thus potentially targetable, host processes upon which the parasite is reliant. Targeting host factors takes the evolutionary control away from the parasite, reducing the probability of resistance emergence. In all, this proposal seeks to address the persistent problem of antimalarial drug resistance development by evaluating two new strategies for discovering resistance-resistant drug targets.