Functional analysis of epigenetic regulators of malaria blood-stage proliferation and transmission

项目摘要 疟疾仍然是一种主要的全球性传染病，主要影响生活在资源贫乏环境中的人们。 人感染恶性疟原虫寄生虫导致显著的发病率和死亡率。毒品- 抗药性不断破坏抗疟疗法的效用。必须确定新的 我们将继续努力，以确保控制和根除疟疾工作取得成功。此外，呼吁防治疟疾 根除需要了解寄生虫传播的决定因素。 恶性疟原虫寄生虫利用表观遗传机制来调节细胞内的关键过程， 寄生虫的生命周期，包括在无性繁殖期间持续存在和发病的抗原变异， 以及通过寄生生命周期转换为性发育以进行传播。我们所获得 遗传学证据表明，II类组蛋白脱乙酰基酶（HDAC）是恶性疟原虫生长所必需的， 无性循环我们假设这些恶性疟原虫HDAC调节细胞中的离散和关键功能， 寄生虫的无性和有性生物学。这些机制的抑制将导致P的特异性杀伤。 恶性疟原虫无性和/或有性形式。 在这个建议中，我们将确定这些酶的基本功能，这些酶负责无性繁殖。 增殖、转化为性阶段和性发育以进行传播。我们将使用反向 在无性和有性阶段的突变寄生虫的表型分析的遗传学方法。我们将 使用批量和单细胞转录组学的组合来探测HDAC功能的机制基础， 表观基因组学最后，我们将通过剖析HDAC的功能来揭示其确切的作用机制。 它们的特定酶结构域，使用生物化学和化学遗传学方法。所有这些 研究将有助于阐明这些关键的表观遗传调节因子在寄生虫生物学中的功能， 无性繁殖的毒力和性发育的传播，验证和表征这些 分子作为寄生虫内治疗发展的新靶点。

PROJECT SUMMARY Malaria remains a major global infectious disease, largely affecting people living in resource poor environments. Infection of humans with Plasmodium falciparum parasites results in significant morbidity and mortality. Drug- resistance is constantly undermining the usefulness of antimalarial regimens. It is imperative to identify new antimalarial drug targets to ensure the success of control and eradication efforts. Further, the call for malaria eradication requires an understanding of the determinants of parasite transmission. Plasmodium falciparum parasites utilize epigenetic machinery for the regulation of key processes within the parasitic life-cycle, including antigenic variation for persistence and pathogenesis during asexual proliferation, as well as for switching to sexual development for transmission through the parasitic life-cycle. We have obtained genetic evidence that the Class II histone deacetylases (HDACs) are essential for P. falciparum growth during the asexual cycle. We hypothesize that these P. falciparum HDACs regulate discrete and critical functions in the asexual and sexual biology of the parasite. Inhibition of these mechanisms will result in the specific killing of P. falciparum asexual and/or sexual forms. In this proposal, we will identify the essential functions of these enzymes that are responsible for asexual proliferation, conversion to the sexual stage, and sexual development for transmission. We will use a reverse genetics approach for the phenotypic analysis of mutant parasites in the asexual and sexual stages. We will probe the mechanistic basic of HDAC functions using a combination of bulk and single cell transcriptomics and epigenomics. Finally, we will uncover the precise mechanisms of action of the HDACs by dissecting the functions of their specific enzymatic domains, using biochemical and chemical genetic approaches. Together, these studies will serve to elucidate the functions of these critical epigenetic regulators in parasite biology relevant to asexual proliferation for virulence and sexual development for transmission, validating and characterizing these molecules as new targets for therapeutic development within the parasite.