Plasmodium cynomolgi as a model for P. vivax.

食蟹猴疟原虫作为间日疟原虫的模型。

• 批准号: 8290557
• 负责人: MARY R GALINSKI
• 金额: $ 17.6万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8290557
• 关键词: Address; Animal Model; Anopheles Genus; Applied Genetics; Area; Basic Science; Biochemical; Biochemistry; Biological; Biological Models; Biology; Blood; Cataloging; Catalogs; Caveolae; Cebidae; Cell membrane; Cells; Cellular biology; Cleaved cell; Color; Complex; Culicidae; Cytoplasm; Dependence; Development; Disease; Erythrocytes; Face; Future; Genetic; Genetic Transformation; Goals; Human; Immunology; In Vitro; Infection; Intervention; Investigation; Knowledge; Laboratories; Life Cycle Stages; Liver; Location; Macaca mulatta; Malaria; Membrane; Microscopy; Modality; Modeling; Morphology; Parasitemia; Parasites; Pathway interactions; Performance; Phylogenetic Analysis; Pilot Projects; Plasmodium cynomolgi; Plasmodium falciparum; Plasmodium vivax; Procedures; Proteins; Proteomics; Regulation; Research; Resources; Reticulocytes; Rodent; Sporozoites; Staging; Surface; System; Technology; Tertiary Protein Structure; Therapeutic; Time; Transfection; Vacuole; Vesicle; biological research; experience; feeding; in vivo; in vivo Model; interest; model development; novel; pathogen; programs; protein transport; research study; success; tool; trafficking; vaccine development

DESCRIPTION (provided by applicant): This R21 proposal focuses on developing in vivo and in vitro Plasmodium cynomolgi model systems to advance cell biological and biochemical research on P. vivax, a major widespread human malaria pathogen. In particular, the goal is to enable the routine application of transfection technologies that will address hypothesis-driven questions regarding the unique biology held in common between P. vivax and P. cynomolgi, which is not found in the other major human malaria, P. falciparum or any of the popular rodent malaria models. This biology includes, among many other differences, the development and reactivation of dormant liver-stage forms known as hypnozoites, and the synthesis and functioning of numerous caveolae vesicle complexes (CVCs) and other membrane structures in infected red blood cells (iRBCs). While P. vivax cannot be cultured continuously in vitro, to facilitate such experimentation, P. cynomolgi blood-stage parasites can be easily manipulated genetically in vivo and ex vivo from rhesus monkey infections where large quantities of parasites can be obtained as well as be adequately propagated and experimentally manipulated by in vitro culture for several days to weeks. The phylogenetic close kinship and nearly identical basic morphology and biology shared by these two species strongly support the proposed utility of P. cynomolgi model systems. No rodent malaria model or P. falciparum culture system offers any biology that mimics the unique biology of these species, and thus they cannot serve as model systems to forward this research. In Aim 1 we will focus on attaining transformed P. cynomolgi parasites that constitutively throughout the life-cycle or at specific developmental time points express single or dual-color fluorescent tags, as the first step for future studies aiming to identify, purify and investigate specific life cycle stages, with a primary interest at this time on the development and activation of the elusive hypnozoite stage. In Aim 2, transformed parasites will be developed to study the trafficking machinery, pathways and unique membrane structures produced by P. vivax and P. cynomolgi blood-stage parasites, which include the CVCs, extensive networks of cleft membranes and novel proteins catalogued by proteomics, immunochemical studies and microscopy. One specific question to be addressed is how and when the PHIST81-95 protein traffics from the parasite beyond the parasitophorous vacuole membrane to its RBC cytosolic location on the cytoplasmic face of the CVCs. For this, the Conditional Aggregation Domain (CAD) protein regulation system will be applied. Critically, P. cynomolgi parasites provide a superior model for the proposed studies, and development of this model will enable research on initial hypothesis-driven questions posed here and ongoing research to understand and define biological and biochemical targets of intervention for P. vivax.

描述（由申请人提供）：该R21提案着重于开发体内和体外疟原虫模型模型系统，以推动细胞生物学和生物化学研究对Vivax的生物学研究，这是一种主要的广泛的人类疟疾病原体。特别是，目的是实现转染技术的常规应用，该技术将解决有关假设驱动的问题，这些问题涉及Vivax和P. cynomolgi之间共同的独特生物学，这在其他主要的人类疟疾，恶性疟原虫或任何流行的啮齿动物疟疾模型中都找不到。该生物学包括在许多其他差异中，还包括休眠肝阶段的发展和重新激活，称为催眠者，以及在受感染的红细胞（IRBC）中的许多小窝囊泡复合物（CVC）和其他膜结构的合成和功能。虽然不能在体外连续培养疟原虫，以促进这种实验，但可以轻松地从遗传上进行遗传和从恒河猴感染中进行遗传操纵，从而可以通过大量的寄生虫进行体内感染，并且可以通过经过数量的传播和实验性地进行几个星期的培养。这两个物种共享的系统发育近亲亲属和几乎相同的基本形态和生物学强烈支持P. cynomolgi模型系统的拟议实用性。没有啮齿动物疟疾模型或恶性疟原虫培养系统提供任何模仿这些物种独特生物学的生物学，因此它们不能用作转发这项研究的模型系统。在AIM 1中，我们将专注于获得转化的cynomolgi寄生虫，这些寄生虫在整个生命周期或特定的发育时间点上表达单个或双色荧光标签，这是未来研究的第一步，旨在识别，纯化和研究特定的生命周期阶段，并在此时间对特定的生命周期阶段进行主要的兴趣，并在此时间对富有利益的型透明度型均具有激进的影响。在AIM 2中，将开发转化的寄生虫来研究由P. vivax和P. cynomolgi血液阶段寄生虫产生的运输机制，途径和独特的膜结构，其中包括CVC，CLEFT膜的广泛网络，新的蛋白质网络和新型蛋白质由蛋白质学，免疫化学，免疫化学，微化学研究分解。要解决的一个具体问题是，phist81-95蛋白运输是如何以及何时从寄生虫液态膜以外的寄生虫到其在CVC的胞质面上的RBC胞质位置。为此，将应用条件聚集域（CAD）蛋白调节系统。至关重要的是，cynomolgi寄生虫为拟议的研究提供了一个卓越的模型，该模型的发展将使在此处和正在进行的研究中提出的初始假设驱动的问题可以理解和定义生物学疟原虫干预的生物学和生化目标。