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有关的胞内病原体，研究较多。 超过150年。它们是艾滋病患者的机会性病原体，最常见的是导致腹泻， 脑炎、肌炎或结膜炎。在晚期艾滋病患者中，高达30%的患者是病因 慢性腹泻伴消瘦的病例。微孢子虫也被发现在其他地方引起感染 免疫受损的宿主，例如接受过器官移植或免疫的患者 调整疗法。感染现在也在免疫能力强的宿主中被识别，导致 角结膜炎或腹泻。微孢子虫被归类为NIH B类优先病原体和EPA 重要的病原体通过食物和水源传播。除了作为人之外 病原体，这些病原体对农业有重大的经济影响(通过对昆虫的影响 水产养殖和动物(食用、家养和野生动物)。动物感染的范围从 隐蔽的良性感染对壮观的大规模感染会造成广泛的损害，通常会导致死亡 主持人。微孢子虫产生的孢子含有一种独特的入侵细胞器，即极管，它是 生物界已知的最复杂的单细胞形式。极地运动的机制 管子在入侵过程中与宿主细胞相互作用尚不清楚。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以及 极管的组成。我们已经开发了纯化这种结构的技术，鉴定出 极管蛋白(PTP)及其翻译后修饰，并确定了研究这些修饰的方法 蛋白质相互作用。此外，我们的研究已经确定了侵袭性突触和功能角色(S) 在入侵过程中的几个PTP。然而，这个结构中的全部蛋白质和 这些成分在入侵过程中的相互作用仍有待确定。这项研究拨款将雇用 蛋白质组、免疫学和超微结构相结合的研究以表征极管及其 蛋白质相互作用组，以更好地定义和理解入侵机制。此外，高级 将使用显微技术(即冷冻-EM和超分辨率显微镜)来提供洞察力 提供关键信息的极管的三维结构和PTP的排列 关于这一入侵细胞器的组织的基本问题 由传统显微镜来分辨。在其他微生物中，关于入侵的研究为 了解发病机制并寻找新的治疗方法来管理感染。我们有 已经证明，针对各种PTPs的抗血清可以抑制侵袭和感染。我们相信这项研究 在萌发和入侵期间这种细胞器的组成、形成和功能的变化应该提供一种 为制定控制这些重要的艾滋病毒相关病原体的新战略奠定了基础。