Microsporidia: invasion apparatus

微孢子虫：入侵装置

• 批准号: 9199134
• 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/9199134
• 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; Health; 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; Patients; Penetration; Phylogenetic Analysis; Post-Translational Protein Processing; Preparation; Process; Proteins; Proteome; Proteomics; Publishing; 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; Water; base; economic impact; fungus; gastrointestinal; gene function; genome sequencing; insight; interest; novel therapeutic intervention; parasite invasion; pathogen; programs; protein structure; receptor; reproductive; research study; 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 微孢子虫是与真菌相关的重要寄生虫，已被研究了更多。 超过150年。他们在剥削所有动物方面表现出色，从神秘的、良性的 感染到壮观的、大规模的感染，造成广泛的损害，常常导致宿主死亡。 微孢子虫是艾滋病患者的机会致病菌，最常引起腹泻， 脑炎、肌炎或结膜炎。微孢子虫还可引起其他免疫系统感染 受损宿主，例如接受器官移植的患者或接受免疫治疗的患者 调节疗法。它们还能够感染免疫能力强的宿主，最常见的原因是 角结膜炎或腹泻。这些病原生物被列为 NIH B 类重点生物 病原体和 EPA 病原体，因为它们通过食物和水源传播。在 除了作为人类病原体外，微孢子虫病还对农业产生重大经济影响（通过影响 昆虫和养蚕）、水产养殖和动物（食品、家畜和野生动物）。微孢子虫产生 孢子具有独特的入侵机制，即极管，这是最复杂的单细胞细胞之一 生物界已知的形式。极管与宿主细胞相互作用的机制 入侵期间仍未知。我的实验室小组的一个长期研究项目专注于 了解侵袭机制以及极管的结构生物学和组成。我们 开发了这种结构的纯化技术，鉴定了极管蛋白（PTP）和 它们的翻译后修饰以及这些蛋白质如何相互作用。另外，我们的学习也开始了 定义这些蛋白质在极管结构生物学和过程中的功能作用 入侵。然而，该结构中蛋白质的完整补充以及这些蛋白质的相互作用 入侵过程中的成分仍有待确定，在其他微生物的入侵研究中已经提供了 了解发病机制和管理新治疗方法的关键数据 感染。我们已经证明主要的极管蛋白是 O-manosylated，一种翻译后蛋白 参与入侵的修饰，抑制甘露糖的结合可以限制感染，并且 极管蛋白 1 (PTP1) 抗体可以阻断侵袭，证明靶向侵袭细胞器 是限制感染的一种方法。拟议的研究项目将结合蛋白质组学、 免疫学和超微结构研究，以表征极管及其蛋白质相互作用组，以便更好地 定义并研究入侵机制。我们还将评估新发现的极地的能力 管蛋白（PTP4）与宿主细胞成分结合，并使用 PTP4 抗体作为标记来识别 发生侵袭的细胞区域。这将有助于详细的相关微观分析 了解这些病原体的入侵机制。此外，电子显微技术将 用于深入了解构成极管的蛋白质的三维结构 提供有关这种入侵细胞器组织的基本问题的关键信息 这是传统显微镜无法解决的问题。组成、形成和研究 该细胞器在萌发和侵袭过程中的功能应该为细胞的发育提供基础 控制这些重要的寄生原生生物的新策略。