Functional genomic analysis of host determinants of malaria infection

描述(申请人提供)：疟疾是全球儿童死亡的主要原因之一，每年造成数十万儿童死亡。虽然抗疟疾药物在流行地区广泛可用，但由于恶性疟原虫抗药性的迅速发展，这些药物的效用受到了影响。疟疾的所有临床症状都可归因于寄生虫寄生在红细胞内的感染阶段。由于这些细胞是去核的，它们是宿主导向疗法的一个有吸引力的靶点，因为它们可能不太可能产生耐药性。人口遗传学研究支持宿主因素可以调节疟疾感染严重程度的观点，但我们对这些因素的识别知识有限。我们设计了一种基于RNAi的遗传筛选，以发现恶性疟原虫在红细胞中入侵或生长所需的宿主因素。我们的初步筛查已确定~10%的红细胞蛋白质组可能影响疟疾的发病机制。这项提议的总体目标是对第一次筛选中确定的两个最有希望的候选宿主因素进行验证和功能研究，并进行第二次筛选，以中等吞吐量的方式验证和描述其他候选因素。除了拟议的研究，候选人的职业发展目标是发展传染病、分子寄生虫学和红细胞遗传学方面的宿主-病原体相互作用方面的专业知识。这些目标将通过参加寄生虫学、热带医学和生物信息学的正式课程以及参加血液学和寄生虫学研讨会和会议来实现。她将得到寄生虫学资深科学家的直接指导，并将通过与血液学和功能基因组学领域的专家合作，发展红细胞生物学和遗传学方面的专业知识。此外，候选人还将受益于与她的科学咨询委员会的互动，该委员会由多个学科的知名科学家和内科科学家组成，包括寄生虫学、血液学和微生物学。候选人的长期职业目标是获得儿科传染病学术医学系的教职，并获得独立资金，以便继续她对疟疾宿主与病原体相互作用的研究。哈佛大学公共卫生学院、波士顿儿童医院和布罗德研究所提供了丰富的培训环境，提供了完成拟议研究所需的所有资源。波士顿儿童医院致力于候选人的职业发展计划，并保证候选人在获奖期间将有超过80%的受保护时间用于拟议的研究。 项目简介：恶性疟疾是全球儿童发病和死亡的一个重要原因，每年造成60多万人死亡。耐药性阻碍了目前临床使用的以寄生虫为导向的抗疟疾药物的有效性；另一种方法是针对疟疾感染所必需的宿主蛋白。在这里，主持人 恶性疟原虫在红细胞中复制所需的蛋白质将使用基因筛查进行鉴定，并根据它们在宿主-寄生虫相互作用中的作用进行表征。这项工作的结果将最终指导疟疾宿主靶向疗法的合理发展。

DESCRIPTION (provided by applicant): Malaria is one of the leading causes of childhood mortality globally, responsible for the deaths of hundreds of thousands of children per year. While antimalarial drugs are widely available in endemic areas, their utility is compromised by the rapid development of drug resistance by Plasmodium falciparum parasites. All of the clinical symptoms of malaria are attributable to the stage of infection when parasites reside within erythrocytes. Since these cells are enucleated, they represent an attractive target for host-directed therapeutics, as they may be less likely to develop resistance. Population genetic studies support the idea that host factors can modulate the severity of malaria infections, but we have limited knowledge of the identity of such factors. We have designed an RNAi-based genetic screen to discover host factors required for invasion or growth of P. falciparum in erythrocytes. Our primary screen has identified ~10% of the erythrocyte proteome as candidates that may influence malaria pathogenesis. The overall goals of this proposal are to perform validation and functional studies on two of the most promising candidate host factors identified in the primary screen, as well as to perform secondary screens to validate and characterize the other candidates in a medium-throughput fashion. Along with the proposed research, the candidate's career development goals are to develop expertise in host-pathogen interactions in infectious diseases, molecular parasitology and erythrocyte genetics. These goals will be achieved by taking formal courses in Parasitology, Tropical Medicine, and Bioinformatics, as well as by attending Hematology and Parasitology seminars and conferences. She will receive direct mentorship from senior scientists in Parasitology, and will develop expertise in erythrocyte biology and genetics by collaborating with experts in the Hematology and functional genomics fields. In addition, the candidate will benefit from interactions with her Scientific Advisory Committee, which is composed of established scientists and physician-scientists from several disciplines, including Parasitology, Hematology, and Microbiology. The candidate's long-term career goals are to obtain a faculty position in academic medical department of Pediatric Infectious Diseases and to secure independent funding in order to continue her research on the host-pathogen interactions in malaria. The Harvard School of Public Health, Boston Children's Hospital, and The Broad Institute offer a rich training environment with all of the resources necessary to complete the proposed research. Boston Children's Hospital is committed to the candidate's career development plan and have assured that the candidate will have more than 80% protected time for the proposed research during the award period. PROJECT NARRATIVE: Plasmodium falciparum malaria is a significant cause of morbidity and mortality among children globally, responsible for over 600,000 deaths per year. Drug resistance hampers the effectiveness of parasite-directed antimalarial drugs currently in clinical use; an alternative approach is to target host proteins essential for malaria infection. Here, host proteins required for P. falciparum replication in erythrocytes will be identified using a genetic screen and characterized in terms of their roles in the host-parasite interaction. The results of this work will ultimately guide the rational development of host-targeted therapeutics for malaria.