Investigating host kinase modulation of erythrocyte deformability during Plasmodium falciparum invasion

项目总结/摘要 疟疾是一种由专性细胞内疟原虫寄生虫引起的毁灭性疾病， 仅在2017年，就有2.19亿人感染了寄生虫，43.5万人感染了 杀了迄今为止，每一种抗疟药物都被耐药性病例抵消，这突出表明， 迫切需要新的药物靶点。靶向宿主因子是一种未充分探索的替代方法， 增加了疟疾抗药性的障碍。这一战略有可能发现所需的目标， 由于宿主需求的重叠，疟原虫有多种物种。此外，许多疟原虫宿主因子 已经是FDA批准的药物靶点，为药物再利用提供了令人信服的理由。 在疟原虫入侵期间，红细胞（RBC）不是一个被动的旁观者;越来越多的证据表明 表明红细胞宿主在寄生虫附着和入侵期间被增选以修改其自身的变形性， 降低寄生虫进入的能量成本。这些变化是由RBC细胞骨架的磷酸化驱动的。 宿主RBC激酶TRPM 7、酪蛋白激酶II（CKII）和Syk是FDA批准的药物靶标， 在恶性疟原虫侵袭期间磷酸化RBC细胞骨架。我假设疟原虫的信号 裂殖子侵入时红细胞骨架通过这些激酶调节红细胞变形性 方便寄生虫进入我的初步数据显示恶性疟原虫的入侵效率 在靶向TRPM 7、CSNK 2A 1和SYK激酶的个体RBC敲除中。 利用整合的遗传学、化学遗传学和磷酸蛋白质组学方法，该提案旨在 研究TRPM 7、CKII和Syk激酶在恶性疟原虫侵袭期间对RBC变形贡献， 验证靶向这些激酶的小分子抑制剂，并阐明寄生虫刺激和下游 恶性疟原虫入侵过程中介导宿主激酶信号转导的效应子。如果成功，这些研究 将揭示更多关于疟原虫入侵的基础生物学，确定有希望的药物治疗宿主靶点， 重新利用血液阶段疟疾，并促进广谱抗疟化合物的开发 增加了抗药性的屏障。最后，由于许多其他血液学疾病受 这些激酶包括巨血小板减少症和动脉纤维化（TRPM 7），遗传性球形红细胞增多症和 链球菌介导的溶血（Syk）和几种血液恶性肿瘤（CKII）-收集的见解 从这些功能分析将有利于寄生虫学和血液学社区。 这项研究将作为我在哈佛T. H.的博士论文的一部分进行。 公共卫生学院。在此期间，我将获得和加强技术技能，包括化学 遗传学、寄生虫学和蛋白质组学;以及微调数据分析、科学交流、指导和 专业发展技能需要成为一个独立的研究科学家在行业。

PROJECT SUMMARY/ABSTRACT Malaria is a devastating disease caused by obligate intracellular Plasmodium parasites that infect millions of people every year: In 2017 alone, 219 million people were infected by the parasite and 435,000 people were killed. Every antimalarial deployed to date has been counteracted by cases of drug resistance, highlighting a desperate need for novel drug targets.Targeting host factors is an underexplored alternative approach to increasing barriers to drug resistance in malaria. This strategy has the potential to uncover targets required for multiple Plasmodium species due to overlaps in host requirements. Additionally, many Plasmodium host factors are already FDA approved drug targets, making a compelling case for drug repurposing. The red blood cell (RBC) is not a passive bystander during Plasmodium invasion; mounting evidence suggests the RBC host is co-opted into modifying its own deformability during parasite attachment & invasion to reduce the energy costs of parasite entry. These changes are driven by phosphorylation of the RBC cytoskeleton. Host RBC kinases TRPM7, Casein Kinase II (CKII), and Syk are FDA approved drug targets postulated to phosphorylate the RBC cytoskeleton during P. falciparum invasion. I hypothesize that Plasmodium signaling to the RBC cytoskeleton during merozoite invasion occurs through these kinases to modulate RBC deformability and facilitate parasite entry. My preliminary data indicates that P. falciparum invasion efficiency is compromised in individual RBC knockouts targeting TRPM7, CSNK2A1, and SYK kinases. Using an integrated genetic, chemical genetic, and phosphoproteomics approach, this proposal seeks to investigate TRPM7, CKII, and Syk kinase contributions to RBC deformation during P. falciparum invasion, validate small molecule inhibitors targeting these kinases, and elucidate parasite stimuli and downstream effectors mediating host kinase signal transduction during P. falciparum invasion. If successful, these studies will uncover more about the basic biology of Plasmodium invasion, identify promising host targets for drug repurposing in blood stage malaria, and facilitate the development of broad spectrum antimalarial compounds with increased barriers to drug resistance. Finally, as many additional hematological disorders are affected by these kinases—including macrothrombocytopenia & arterial fibrosis (TRPM7), hereditary spherocytosis & Streptococcus-mediates hemolysis (Syk), and several hematological malignancies (CKII)—insights gleaned from these functional analyses will benefit both parasitology and hematology communities. The proposed research will be carried out as part of my doctoral dissertation studies at the Harvard T.H. Chan School of Public Health. During this fellowship I will acquire & strengthen technical skills including chemical genetics, parasitology, and proteomics; and fine-tune data analysis, scientific communication, mentorship, and professional development skills needed to become an independent research scientist in industry.