Functional investigation of a novel and essential subcellular compartment in Plasmodium falciparum transmission stage parasites

恶性疟原虫传播阶段寄生虫中新型重要亚细胞区室的功能研究

• 批准号: 10584525
• 负责人: JEFFREY D DVORIN
• 金额: $ 75.31万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-04 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10584525
• 关键词: 3-Dimensional; 5 year old; Biogenesis; Biophysics; Blood; Cell membrane; Cellular Morphology; Cessation of life; Child; Complex; Culicidae; Cytoskeleton; Defect; Development; Electron Microscopy; Epitopes; Erythrocytes; Female; Filament; Future; Gene Proteins; Genes; Goals; Human; Image; Immunofluorescence Immunologic; Infection; Ingestion; Intermediate Filaments; Investigation; Ions; Knock-out; Label; Life Cycle Stages; Malaria; Measures; Membrane; Microfabrication; Microtubules; Molecular; Morbidity - disease rate; Morphology; Multiprotein Complexes; Names; Parasites; Pathway interactions; Plasmodium falciparum; Platinum; Population; Process; Proteins; Scanning Electron Microscopy; Series; Shapes; Techniques; Testing; Transgenic Organisms; Transmission Electron Microscopy; Vesicle; Visualization; Width; asexual; improved; knock-down; male; member; mortality; novel; novel strategies; resilience; reverse genetics; transmission process

PROJECT SUMMARY Malaria is an important cause of illness and death worldwide, with most of these deaths resulting from Plasmodium falciparum infection. Successful completion of the P. falciparum life cycle and infection of a new human host requires transmission. During the asexual blood stage in human red blood cells, a small population of parasites differentiates into transmission forms known as gametocytes. These gametocytes can complete the sexual stage of the parasite life cycle following ingestion by a mosquito. Gametocyte maturation in human red blood cells occurs over 10-12 days and is associated with major changes in cellular morphology and rigidity. A newly discovered protein PfBLEB (for Baso-Lateral Expansion Boundary) is essential for mature gametocyte formation. In asexual parasites, PfBLEB is part of the basal complex, a ring-like multi-protein complex at the leading edge of the inner membrane complex. While PfBLEB is dispensable for both asexual replication and gametocyte commitment, it is essential for gametocyte development. PfBLEB-knockdown or knockout gametocytes arrest during maturation and are non- transmissible. Furthermore, the PfBLEB-deficient gametocytes have gross morphologic changes with defects in major cytoskeletal features of the maturing transmission-stage parasite, including the inner membrane complex and subpellicular microtubules. In gametocytes with normal PfBLEB expression, PfBLEB defines a new subcellular compartment within the parasite, demarcating the regions of the parasite plasma membrane that are devoid of the underlying inner membrane complex. The PfBLEB-compartment is essential for gametocyte development, but the function and protein constituents of this newly discovered subcellular compartment remain unknown. The goal of the current application is to define and genetically evaluate the protein components of the PfBLEB compartment and to understand the functional defects in PfBLEB-deficient gametocytes. The first aim will utilize proximity labeling and reverse genetics to explore the PfBLEB-containing compartment. The second aim will utilize multiple imaging and microfabrication techniques to gain a functional understanding of what processes are abnormal in PfBLEB-deficient gametocytes. Together, the proposed studies will add a new layer to our molecular understanding of gametocyte development in P. falciparum.

项目摘要 疟疾是全球疾病和死亡的重要原因，其中大多数死亡是由 恶性疟原虫感染。成功地完成了恶性疟原虫生命周期和A的感染 新的人类主机需要传输。在人类红细胞的无性血液阶段，一个小 寄生虫的种群将被称为配子细胞的传输形式区分开。这些配子细胞 蚊子摄入后，可以完成寄生虫生命周期的性阶段。配子细胞 人类红细胞的成熟发生在10-12天内，与重大变化有关 细胞形态和僵化。新发现的蛋白质pfbleB（用于基本外侧膨胀边界） 对于成熟的配子细胞形成至关重要。在无性寄生虫中，pfbleB是基底复合物的一部分 在内膜复合物的前缘，环状多蛋白复合物。而pfbleB是 对于无性复制和配子细胞的承诺，对配子细胞至关重要 发展。 pfbleb-knockdown或敲除配子细胞在成熟过程中停滞，而不是 - 可传播。此外，pfbleB缺陷的配子细胞与形态学发生了严重的变化， 成熟传输阶段寄生虫的主要细胞骨架特征的缺陷，包括内部 膜络合物和细胞微管。 在具有正常pfbleB表达的配子细胞中，pfbleB定义了一个新的亚细胞隔室 寄生虫，划定缺乏基础内部的寄生虫质膜区域 膜络合物。 PfbleB-Compartment对于配子细胞的开发至关重要，但是功能 该新发现的亚细胞室的蛋白质成分仍然未知。目标的目标 当前的应用是定义和遗传评估PfbleB室的蛋白质成分 并了解pfbleB缺陷的配子细胞中的功能缺陷。第一个目标将利用接近性 标记和反向遗传学以探索含PfbleB的隔室。第二个目标将利用 多个成像和微加工技术，以获得对哪些过程的功能理解 PfbleB缺陷的配子细胞异常。拟议的研究一起将为我们的 对恶性疟原虫的配子细胞发育的分子理解。