Microsporidia: invasion apparatus

微孢子虫：入侵装置

• 批准号: 9273478
• 负责人: Louis M. Weiss
• 金额: $ 41.75万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-16 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9273478
• 关键词: Acquired Immunodeficiency Syndrome; Agriculture; Animal Feed; Animals; Antibodies; Aquaculture; Area; Benign; Binding; Bioinformatics; Biological; Biological Preservation; Birds; Categories; Cell membrane; Cells; Cellular Structures; Cellular biology; Cessation of life; Collaborations; Comparative Study; Complement; Complex; Conjunctivitis; Data; Development; Diarrhea; Economics; Electrons; Encephalitis; Encephalitozoon hellem; Enterocytozoon bieneusi; Epitopes; Fishes; Food; Genetic; Genome; Germination; HIV; Host-Parasite Relations; Human; Human Genome; Immune; Immunization; Immunoelectron Microscopy; Immunologics; In Vitro; Infection; Insecta; Invaded; Invertebrates; Investigation; Keratoconjunctivitis; Laboratories; Laboratory Research; Location; Mammals; Manuscripts; Methods; Microbe; Microscopic; Microscopy; Microsporidia; Microsporidiosis; Molecular; Molecular Analysis; Morphology; Mus; Myositis; Nematoda; Nervous system structure; Nosema; Nosema corneum; Organ Transplantation; Organelles; Organism; Parasites; Pathogenesis; Pathogenicity; Patients; Penetration; Phylogenetic Analysis; Post-Translational Protein Processing; Preparation; Process; Proteins; Proteome; Proteomics; Publishing; Receptor Cell; Recombinants; Reproduction spores; Research; Research Project Grants; Resources; Respiratory Muscles; Role; Septata intestinalis; Sister; Site; Soil; Source; Structure; System; Techniques; Transfection; Tube; United States National Institutes of Health; Validation; Vitronectin; Water; base; economic impact; experimental study; fungus; gastrointestinal; gene function; insight; interest; novel therapeutic intervention; parasite invasion; pathogen; pathogen genome; programs; protein structure; public health relevance; receptor; reproductive; sample fixation; structural biology; sugar; tool; transcriptome sequencing

ABSTRACT Microsporidia are remarkable parasites related to the Fungi that have been studied for more than 150 years. They are remarkable in their exploitation of all animals ranging from cryptic, benign infections to spectacular, massive infections that cause extensive damage and often death of the host. Microsporidai are opportunistic pathogens in patients with AIDS most commontly causing diarrhea, encephalitis, myositis, or conjunctivitis. Microsporidia can also cause infections in other immune compromised hosts, such as patients who have undergone organ transplantation or those on immune modulating therapies. They are also capable of infecting immune competent hosts most commonly causing keratoconjunctivitis or diarrhea. These pathogenic organisms are classified as NIH category B priority pathogens and EPA pathogens of interest as they are transmitted by both food and water sources. In addition to being human pathogens, microsporidiosis has major economic impacts on agriculture (via effects on insects and sericulture), aquaculture and animals (food, domestic and wildlife). Microsporidia produce spores with a unique invasion mechanism, the polar tube, that is one of the most complex single celled forms known in the biological world. The mechanism by which the polar tube interacts with the host cell during invasion is still unknown. A long standing research program in my laboratory group is focused on understanding the mechanism of invasion and the structural biology and composition of the polar tube. We have developed techniques for the purificaiton of this structure, identified polar tube proteins (PTPs) and their post translational modifications and how these proteins interact. Futhermore, our studies have begun to define the functional roles of these proteins in the structural biology of the polar tube and the process of invasion. However, the full complement of proteins in this structure and the interactions of these components during invasion remain to be determined, In other microbes studies on invasion have provided key data for understanding pathogenesis and for new therapeutic approaches to the management of infections. We have demonstrated that the major polar tube protein is O-manosylated, a post translational modification that is involved in invasion, that inhibiting binding of manose can limit infection, and that antibody to polar tube protein 1 (PTP1) can block invasion demonstrate that targeting the invasion organelle is a way to limit infection. The proposed research project will employ a combination of proteomic, immunologic and ultrastructural studies to characterize the polar tube and its protein interactome to better define and study the mechanism of invasion. We will also evaluate the ability of a newly identified polar tube protein (PTP4) to bind to host cell components and use antibody to PTP4 as a marker to identify the area of the cell at which invasion is occurring. This will facilitate a detailed correlated microscopic analysis of the mechanism of invasion by these pathogens. Furthermore, electron microscopic techniques will be employed to provide insight into the three dimenstional structure of the proteins making up the polar tube providng critical information on fundamental questions concerning the organization of this invasion organelle that have not been able to be resolved by traditional microscopy. Studies of the composition, formation and function of this organelle during germination and invasion should provide a basis for the development of new strategies for control of these important parasitic protists.

摘要微孢子虫是与真菌有关的显着寄生虫，这些寄生虫已被研究以获取更多 超过150年。他们对所有动物的剥削都很出色，从隐秘，良性 引起壮观，大规模感染的感染，会造成广泛的损害和常常死亡。 微孢子虫是艾滋病患者的机会性病原体，导致腹泻， 脑炎，肌炎或结膜炎。微孢子虫也可能引起其他免疫感染 受损的宿主，例如接受器官移植的患者或免疫的患者 调节疗法。他们还能够感染最常见的免疫能力宿主 角膜结膜炎或腹泻。这些致病生物被归类为NIH类别B的优先级 病原体和EPA病原体是食物和水源传播的。在 除了人类病原体外，微孢子虫病对农业有重大的经济影响（通过影响 关于昆虫和蚕桑），水产养殖和动物（食物，家庭和野生动植物）。微孢子虫生产 具有独特入侵机制的孢子，即极性管，这是最复杂的单元格之一 在生物世界中已知的形式。极性管与宿主细胞相互作用的机制 在入侵期间仍然未知。我的实验室小组的长期研究计划的重点是 了解侵袭机制以及极性管的结构生物学和组成。我们 已经开发了该结构的净化技术，确定的极地管蛋白（PTP）和 他们的后翻译修饰以及这些蛋白质如何相互作用。 futhermore，我们的研究已经开始 定义这些蛋白质在极化管的结构生物学中的功能作用和 入侵。但是，这种结构中蛋白质的完整补体以及这些结构的相互作用 入侵期间的成分仍有待确定，在其他有关侵袭的微生物研究中 理解发病机理和新治疗方法的关键数据 感染。我们已经证明了主要的极性管蛋白是o型蛋白，后翻译后 涉及入侵的修改，抑制Manose的结合可以限制感染，并且 极性管蛋白1（PTP1）的抗体可以阻止侵袭，表明针对入侵器官 是限制感染的一种方式。拟议的研究项目将采用蛋白质组学的组合 免疫学和超微结构研究以表征极化管及其蛋白质相互作用以更好 定义和研究侵袭机制。我们还将评估新确定的极地的能力 管蛋白（PTP4）与宿主细胞成分结合并使用与PTP4的抗体作为标记来识别 发生侵袭的细胞区域。这将促进详细的相关显微镜分析 这些病原体的入侵机制。此外，电子显微镜技术将是 用于洞悉构成极性管的蛋白质的三个尺寸结构 提供有关该入侵组织组织组织的基本问题的关键信息 传统显微镜无法解决的。研究组成，形成和 在发芽和入侵期间，该细胞器的功能应为发展提供基础 控制这些重要的寄生虫生物的新策略。