Microsporidia: invasion apparatus

微孢子虫：入侵装置

• 批准号: 10619448
• 负责人: Louis M. Weiss
• 金额: $ 42万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 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 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10619448
• 关键词: Acquired Immunodeficiency Syndrome; Agriculture; Animals; Antibodies; Aquaculture; Benign; Bioinformatics; Biological; Birds; Category B pathogen; Cell membrane; Cells; Cellular biology; Cessation of life; Chronic diarrhea; Classification; Collaborations; Comparative Study; Complement; Complex; Conjunctivitis; Cryoelectron Microscopy; Data; Development; Diarrhea; Economics; Encephalitis; Encephalitozoon hellem; Enterocytozoon bieneusi; Epitopes; Etiology; Fishes; Food; Genetic; Genome; Germination; Grant; HIV; HIV Infections; Human; Human Genome; Immune; Immune Sera; Immunization; Immunocompetent; Immunocompromised Host; Immunoelectron Microscopy; Immunologics; In Vitro; Infection; Insecta; Invaded; Invertebrates; Investigation; Keratoconjunctivitis; Laboratories; Laboratory Research; Mammals; Methods; Microbe; Microscopic; Microscopy; Microsporidia; Microsporidiosis; Molecular; Morphology; Mus; Muscle; Myositis; Nematoda; Nervous System; Nosema; Nosema corneum; Organ Transplantation; Organelles; Organism; Pathogenesis; Pathogenicity; Patients; Penetration; Phylogenetic Analysis; Post-Translational Protein Processing; Process; Proteins; Proteome; Proteomics; Publishing; Recombinants; Reproduction spores; Research; Research Project Grants; Role; Scanning Electron Microscopy; Septata intestinalis; Sister; Site; Soil; Source; Structure; Synapses; System; TFRC gene; Techniques; Transfection; Tube; United States National Institutes of Health; Validation; Vero Cells; Water; economic impact; emerging pathogen; experimental study; fungus; gastrointestinal; gene function; genome resource; human pathogen; immunomodulatory therapies; infection management; insight; interest; novel therapeutic intervention; opportunistic pathogen; parasite invasion; pathogen; preservation; programs; public health relevance; reproductive; respiratory; sample fixation; structural biology; sugar; superresolution microscopy; three dimensional structure; tool; transmission process; wasting; Acquired Immunodeficiency Syndrome; Agriculture; Animals; Antibodies; Aquaculture; Benign; Bioinformatics; Biological; Birds; Category B pathogen; Cell membrane; Cells; Cellular biology; Cessation of life; Chronic diarrhea; Classification; Collaborations; Comparative Study; Complement; Complex; Conjunctivitis; Cryoelectron Microscopy; Data; Development; Diarrhea; Economics; Encephalitis; Encephalitozoon hellem; Enterocytozoon bieneusi; Epitopes; Etiology; Fishes; Food; Genetic; Genome; Germination; Grant; HIV; HIV Infections; Human; Human Genome; Immune; Immune Sera; Immunization; Immunocompetent; Immunocompromised Host; Immunoelectron Microscopy; Immunologics; In Vitro; Infection; Insecta; Invaded; Invertebrates; Investigation; Keratoconjunctivitis; Laboratories; Laboratory Research; Mammals; Methods; Microbe; Microscopic; Microscopy; Microsporidia; Microsporidiosis; Molecular; Morphology; Mus; Muscle; Myositis; Nematoda; Nervous System; Nosema; Nosema corneum; Organ Transplantation; Organelles; Organism; Pathogenesis; Pathogenicity; Patients; Penetration; Phylogenetic Analysis; Post-Translational Protein Processing; Process; Proteins; Proteome; Proteomics; Publishing; Recombinants; Reproduction spores; Research; Research Project Grants; Role; Scanning Electron Microscopy; Septata intestinalis; Sister; Site; Soil; Source; Structure; Synapses; System; TFRC gene; Techniques; Transfection; Tube; United States National Institutes of Health; Validation; Vero Cells; Water; economic impact; emerging pathogen; experimental study; fungus; gastrointestinal; gene function; genome resource; human pathogen; immunomodulatory therapies; infection management; insight; interest; novel therapeutic intervention; opportunistic pathogen; parasite invasion; pathogen; preservation; programs; public health relevance; reproductive; respiratory; sample fixation; structural biology; sugar; superresolution microscopy; three dimensional structure; tool; transmission process; wasting

ABSTRACT Microsporidia are intracellular pathogens related to the Fungi that have been studied for more than 150 years. They are opportunistic pathogens in patients with AIDS, most commonly causing diarrhea, encephalitis, myositis, or conjunctivitis. In patients with advanced AIDS they have been etiologic in up to 30% of cases of chronic diarrhea with wasting. Microsporidia have also been found to cause infections in other immunocompromised hosts, such as patients who have undergone organ transplantation or those on immune modulating therapies. Infections are now also being recognized in immune competent hosts causing either keratoconjunctivitis or diarrhea. Microsporidia 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, these pathogenic organisms have major economic impacts on agriculture (via effects on insects and sericulture), aquaculture and animals (food, domestic and wildlife). Infections in animals range from cryptic, benign infections to spectacular, massive infections that cause extensive damage and often death of the host. Microsporidia produce spores containing a unique invasion organelle, the polar tube, which is one of the most complex single celled forms known in the biological world. The mechanism by which the polar tube interacts with its 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 purification of this structure, identified polar tube proteins (PTPs) and their post translational modifications, and defined methods to study how these proteins interact. Furthermore, our investigations have defined the invasion synapse and the functional role(s) of several PTPs in 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. This research grant will employ a combination of proteomic, immunologic and ultrastructural studies to characterize the polar tube and its protein interactome to better define and understand the mechanism of invasion. Furthermore, advanced microscopic techniques (i.e. cryo-EM and super resolution microscopy) will be employed to provide insight into the three dimensional structure of the polar tube and arrangement of PTPs providing critical information on fundamental questions concerning the organization of this invasion organelle that have not been able to be resolved by traditional microscopy. In other microbes studies on invasion have provided critical data for understanding pathogenesis and for new therapeutic approaches to the management of infections. We have already demonstrated that antisera to various PTPs can inhibit invasion and infection. We believe that 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 HIV-associated pathogens.

摘要微孢子虫是与真菌有关的细胞内病原体 超过150年。它们是艾滋病患者的机会性病原体，最常见的是腹泻， 脑炎，肌炎或结膜炎。在患有晚期辅助的患者中，他们的病因多达30％ 慢性腹泻的病例。还发现微孢子虫引起其他人的感染 免疫功能低下的宿主，例如接受器官移植的患者或免疫的患者 调节疗法。现在，在免疫胜任的宿主中也认识到感染，从而导致感染 角膜结膜炎或腹泻。微孢子虫被归类为NIH类B类优先病原体和EPA 感兴趣的病原体是由食物和水源传播的。除了人类 病原体，这些致病生物对农业有重大的经济影响（通过对昆虫的影响 和粒土），水产养殖和动物（食物，家庭和野生动植物）。动物的感染范围从 神秘的，良性的感染引起了壮观的，大规模的感染，这些感染造成广泛损害，并且经常死亡 主人。微孢子虫产生含有独特入侵细胞器的孢子，极性管，这是一个 在生物世界中已知的最复杂的单细胞形式。极地的机制 在入侵期间，管与宿主细胞相互作用仍然未知。我的一项长期研究计划 实验室小组的重点是了解入侵机制和结构生物学的机制和 极性管的组成。我们已经开发了纯化该结构的技术， 极性管蛋白（PTP）及其后翻译后修改，并定义了研究这些方法的方法 蛋白质相互作用。此外，我们的研究定义了入侵突触和功能作用 在入侵过程中的几个PTP。但是，该结构中的蛋白质的完整补体和 在入侵期间这些成分的相互作用仍有待确定。这项研究赠款将采用 蛋白质组学，免疫学和超微结构研究的结合，以表征极性管及其 蛋白质相互作用，以更好地定义和理解侵袭机制。此外，先进 将采用显微镜技术（即冷冻EM和超级分辨率显微镜）来提供洞察力 进入极性管的三维结构和PTP的排列，提供关键信息 关于该入侵组织组织组织的基本问题 通过传统显微镜解决。在其他微生物研究中，有关入侵的研究为 了解发病机理和新的治疗方法来管理感染。我们有 已经证明了对各种PTP的抗血清可以抑制入侵和感染。我们认为研究 在发芽和入侵期间，该细胞器的组成，形成和功能应提供 开发新策略以控制这些重要的HIV相关病原体的基础。