Microsporidia: invasion apparatus

微孢子虫：入侵装置

• 批准号: 10324040
• 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/10324040
• 关键词: Acquired Immunodeficiency Syndrome; Agriculture; Animal Feed; Animals; Antibodies; Aquaculture; Benign; Bioinformatics; Biological; Birds; Category B pathogen; Cell membrane; Cells; Cellular biology; Cessation of life; Chronic diarrhea; 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; 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; Myositis; Nematoda; Nervous system structure; 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; Resolution; Respiratory Muscles; Role; Scanning; 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; pathogen genome; preservation; programs; public health relevance; reproductive; sample fixation; structural biology; sugar; three dimensional structure; tool; 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 微孢子虫是与真菌相关的细胞内病原体，已被研究了更多。 超过150年。它们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。在晚期艾滋病患者中，高达 30% 的患者是由病因引起的 慢性腹泻伴消瘦的病例。还发现微孢子虫可引起其他疾病的感染 免疫功能低下的宿主，例如接受过器官移植的患者或接受免疫治疗的患者 调节疗法。现在在免疫能力强的宿主中也发现了感染，导致 角结膜炎或腹泻。微孢子虫被列为 NIH B 类优先病原体和 EPA 感兴趣的病原体，因为它们通过食物和水源传播。除了作为人之外 病原体，这些病原生物对农业具有重大经济影响（通过对昆虫的影响） 和养蚕）、水产养殖和动物（食品、家畜和野生动物）。动物感染的范围包括 从隐秘的良性感染到引起广泛损害甚至死亡的壮观的大规模感染 主人。微孢子虫产生的孢子含有一种独特的入侵细胞器，极管，这是一种 生物界已知的最复杂的单细胞形式。极性的机制 管在入侵过程中与其宿主细胞相互作用仍然未知。我的一项长期研究计划 实验室小组致力于了解入侵机制和结构生物学 极管的组成。我们开发了纯化该结构的技术，确定了 极管蛋白 (PTP) 及其翻译后修饰，并定义了研究这些蛋白如何发挥作用的方法 蛋白质相互作用。此外，我们的研究还定义了入侵突触及其功能作用 入侵过程中的几个PTP。然而，该结构中蛋白质的完整补充和 这些成分在入侵过程中的相互作用仍有待确定。这项研究补助金将雇用 结合蛋白质组学、免疫学和超微结构研究来表征极管及其 蛋白质相互作用组以更好地定义和理解入侵机制。此外，先进的 将采用显微技术（即冷冻电镜和超分辨率显微镜）来提供见解 深入了解极管的三维结构和 PTP 的排列，提供关键信息 关于有关这种入侵细胞器的组织的基本问题尚未能够 可以通过传统的显微镜来解决。在其他微生物的入侵研究中，为以下方面提供了关键数据： 了解发病机制并寻找治疗感染的新治疗方法。我们有 已经证明各种PTPs的抗血清可以抑制侵袭和感染。我们相信，研究 该细胞器在萌发和侵袭过程中的组成、形成和功能应该提供 制定控制这些重要的艾滋病毒相关病原体的新策略的基础。