Characterizing the Plasmodium falciparum Subpellicular Network

描述恶性疟原虫表膜下网络的特征

• 批准号: 10533135
• 负责人: Ana Karla Cepeda Diaz
• 金额: $ 4.05万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2025-09-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10533135
• 关键词: 3-Dimensional; Antimalarials; Architecture; Biochemical; Biogenesis; Biology; Blood; Cell Shape; Cell membrane; Cells; Cessation of life; Complex; Culicidae; Cytoskeletal Proteins; Cytoskeleton; Daughter; Defect; Development; Disease; Drug Targeting; Electron Microscopy; Erythrocytes; Eukaryota; Family; Filament; Flow Cytometry; Fluorescence-Activated Cell Sorting; Human; Immunofluorescence Immunologic; Individual; Intermediate Filaments; Invaded; Knock-out; Life; Malaria; Mammalian Cell; Maps; Mechanical Stress; Medical Economics; Membrane; Microscopy; Monitor; Morphology; Motor; Names; Organelles; Parasites; Pathway interactions; Pattern; Plasmids; Plasmodium; Plasmodium falciparum; Platinum; Play; Process; Proline; Protein Family; Proteins; Protozoa; Reproduction; Research; Resolution; Role; Stains; Structure; System; Testing; Toxoplasma gondii; Transmission Electron Microscopy; Ursidae Family; Valine; asexual; cell motility; daughter cell; drug development; electron tomography; insight; knock-down; live cell microscopy; network architecture; protein function; reconstruction; rhoptry; scaffold

ABSTRACT The cytoskeleton of Plasmodium spp. is essential for replication, motility, and infectivity in both human and mosquito life stages. P. falciparum, the causative agent of the most severe form of human malaria, leverages a family of cytoskeletal proteins known as the alveolins to meet its diverse needs. These intermediate filament-like proteins are absent outside the Alveolate kingdom, making them attractive drug targets. The alveolins have stage-specific expression patterns and form an intricate lattice which envelops the parasite just below the pellicle, another hallmark of Alveolata consisting of the parasite plasma membrane and inner membrane complex. Remarkable progress has been achieved in characterizing the role of alveolins in P. berghei mosquito stages. However, the functional role of individual alveolins in Plasmodium asexual stages remains unexplored. We have recently demonstrated that the alveolin PfIMC1g is essential for P. falciparum asexual replication, but it remains unclear what function this protein serves within the larger context of the Plasmodium cytoskeleton. Using inducible knockdown (iKD) and knockout (iKO) systems in conjunction with super-resolution and platinum replica electron microscopy, I will characterize the role of alveolin PfIMC1g (PF3D7_0525800) in P. falciparum daughter cell segmentation and red blood cell invasion. This will elucidate its individual role in parasite cell shape and ability to endure mechanical stress. In addition, I will use expansion microscopy to map the other alveolins present in the asexual stages of the parasite, their interactions with each other, and test their essentiality for daughter cell formation through conventional KO and iKO approaches. This research will increase our understanding of the functional role that these building blocks of the P. falciparum SPN serve and determine how they come together to enable its formation, function, and remodeling during segmentation. Together, these insights into intermediate-filament-like cytoskeletal proteins and their organizing principles will bring us one step closer to targeting these proteins with new antimalarial drugs.

抽象的 疟原虫的细胞骨架。对于人类和人类的复制，运动和感染性至关重要 蚊子生活阶段。恶性疟原虫是最严重的人类疟疾形式的致病药物，利用A 被称为肺泡蛋白的细胞骨架蛋白家族，可满足其多样化的需求。这些中间细丝 蛋白质不存在于肺泡王国外，使其成为有吸引力的药物靶标。肺泡有 特定于阶段的表达模式并形成一个复杂的晶格，该晶格将寄生虫在颗粒下方的下方， 肺泡的另一个标志是由寄生虫质膜和内膜复合物组成。 在表征肺泡蛋白在伯格（P. berghei P. berghei蚊子阶段）的作用方面取得了显着进步。 但是，单个肺泡素在疟原虫无性阶段中的功能作用仍未开发。我们有 最近证明，肺泡蛋白PFIMC1G对于恶性疟原虫无性复制至关重要，但仍然是 尚不清楚该蛋白在疟原虫细胞骨架的较大背景下起什么功能。使用 诱导型敲低（IKD）和敲除（IKO）系统以及超分辨率和铂金复制品 电子显微镜，我将表征肺泡蛋白PFIMC1G（PF3D7_0525800）在恶性疟原虫女儿中的作用 细胞分割和红细胞侵袭。这将阐明其在寄生虫细胞形状中的个人作用，并且 能够忍受机械应力。此外，我将使用膨胀显微镜绘制其他肺泡蛋白 出现在寄生虫的无性阶段，它们相互互动，并测试其重要性 通过常规的KO和IKO接近女儿细胞形成。这项研究将增加我们的 了解这些功能作用的这些功能作用，这些构成疟原虫SPN的功能并确定 它们如何聚集在一起，使其在分割过程中的形成，功能和重塑。在一起，这些 洞悉类丝状细胞骨架蛋白及其组织原理将为我们带来一步 更接近使用新的抗疟药来靶向这些蛋白质。