Cytoskeletal control of Leishmania infection

利什曼原虫感染的细胞骨架控制

• 批准号: 8261450
• 负责人: Dawn Marie Wetzel
• 金额: $ 4.24万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8261450
• 关键词: Actins; Address; Adhesions; Antiparasitic Agents; Binding; Biochemical; Biochemical Pathway; Biological Assay; Bite; Cell Lineage; Cell Surface Receptors; Cell surface; Cells; Cellular Structures; Cessation of life; Chemicals; Complex; Cytoskeleton; Data; Defect; Development; Disease; Drug Delivery Systems; Event; Family; Fibroblasts; Fibronectins; Genes; Gleevec; Hematopoietic; Homologous Gene; Human; Imatinib; In Vitro; Infection; Integrin Binding; Integrins; Knockout Mice; Knowledge; Laboratories; Lead; Leishmania; Leishmaniasis; Life; Life Cycle Stages; Link; Mediating; Membrane; Molecular Biology Techniques; Mus; Parasites; Parasitic infection; Pathogenesis; Pathway interactions; Phagocytosis; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Predisposition; Preparation; Process; Protein Kinase; Protein Tyrosine Kinase; Proteins; Research; Resistance development; Sand Flies; Signal Pathway; Signal Transduction; Skin Ulcer; Swelling; System; Testing; Translating; Visceral; Work; base; cell motility; extracellular; human EMS1 protein; improved; in vivo; inhibitor/antagonist; killings; leukemia; macrophage; mouse model; novel; parasite invasion; polymerization; public health relevance; receptor; research study; surface coating; uptake

DESCRIPTION (provided by applicant): Leishmania is a parasite that causes severe skin ulcers or visceral disease in people living in the developing world. Every year, it infects millions of people, and kills over 50,000. Drugs for treating infection by Leishmania are very toxic, and the parasite is developing resistance to them. For these reasons, new drugs are needed to treat infections by this parasite. Leishmania must live inside macrophages to survive in and cause disease within a host. To enter macrophages, Leishmania binds to integrin receptors on the cell surface. The parasite then triggers its own uptake through the process of phagocytosis. The mechanisms that permit cell entry by Leishmania are not well understood. This project aims to characterize the signaling events that translate the binding of cell surface receptors to the reorganization of the cytoskeleton, which is required for internalization of Leishmania. Understanding the mechanisms of cell entry by Leishmania will help explain how this parasite causes disease. Furthermore, if one could disrupt the pathways that lead to invasion of macrophages by Leishmania, the parasite should not be able to survive within a host to cause disease. Therefore, these studies may also suggest new drugs to treat this parasitic infection. Abl family kinases are proteins that transfer signals from extracellular integrin receptors to the cytoskeleton in order to orchestrate cell movement. These kinases are known to activate cortactin, which triggers actin-based cell edge protrusions. HS1 (hematopoietic lineage cell-specific protein 1) is a protein related to cortactin that is found in macrophages. Preliminary work in the laboratory demonstrates that signals from cell surface receptors activate Abl family kinases and stimulate phagocytosis. However, signaling pathways requiring Abl family kinases or HS1 have not been linked to uptake of Leishmania by macrophages. The central hypothesis in this proposal is that Leishmania uses a signaling pathway that requires integrin receptors, Abl family kinases, and HS1 to enter cells and cause disease. Specific Aim 1 addresses whether Abl family kinases mediate cell entry and infection by Leishmania, and if 22 integrins directly bind and activate Abl family kinases. Specific Aim 2 addresses whether Leishmania requires HS1 signaling for cell entry and infection; it also tests whether Abl family kinases activate HS1. In both specific aims, macrophages will be infected with Leishmania to understand the requirement for each of these signaling components for cell entry, and mice will be infected with Leishmania to determine the significance of these proteins in disease. Biochemical assays with purified proteins and molecular biology techniques to disrupt interactions between proteins will also be employed to elucidate the interfaces between key components in this signaling pathway. By identifying whether signaling though integrins, Abl family kinases, and HS1 allows Leishmania to enter cells and cause disease in mice, our understanding of the process of Leishmania infection will improve. PUBLIC HEALTH RELEVANCE: This project seeks to explain how a human parasite called Leishmania gains entry into cells in order to survive. Leishmania infects millions of people every year worldwide, and kills more than 50,000 of them. We hope that if we understand how Leishmania enters cells, new drugs can be developed that block cell entry, and therefore, disease caused by this parasite.

描述（由申请人提供）：利什曼尼亚是一种寄生虫，在发展中国家的人们中会导致严重的皮肤溃疡或内脏疾病。每年，它都会感染数百万人，并杀死50,000多人。利什曼原虫治疗感染的药物非常有毒，寄生虫正在对其产生抗药性。由于这些原因，需要新药来治疗这种寄生虫。利什曼尼亚必须生活在巨噬细胞内，以生存并在宿主内引起疾病。要进入巨噬细胞，利什曼原虫与细胞表面上的整联蛋白受体结合。然后，寄生虫通过吞噬作用触发了自己的吸收。允许利什曼原虫进入细胞的机制尚不清楚。该项目旨在表征将细胞表面受体与细胞骨架重组结合的结合的信号传导事件，这是利什曼尼亚内在化所必需的。了解利什曼原虫的细胞进入机制将有助于解释这种寄生虫如何引起疾病。此外，如果可以破坏导致利什曼尼亚侵袭巨噬细胞的途径，则该寄生虫不应在宿主中生存以引起疾病。因此，这些研究还可能建议新药来治疗这种寄生虫感染。 ABL家族激酶是将信号从细胞外整联蛋白受体转移到细胞骨架的蛋白质，以便编排细胞运动。已知这些激酶会激活皮质素，这会触发基于肌动蛋白的细胞边缘突起。 HS1（造血谱系细胞特异性蛋白1）是与巨噬细胞中发现的蛋白质。实验室的初步工作表明，来自细胞表面受体的信号激活了ABL家族激酶并刺激吞噬作用。但是，需要ABL家族激酶或HS1的信号通路与巨噬细胞对利什曼原虫的摄取没有联系。 该提议中的中心假设是利什曼尼亚使用的信号传导途径需要整合素受体，ABL家族激酶和HS1进入细胞并引起疾病。具体目标1解决了ABL家族激酶是否介导利什曼原虫的细胞进入和感染，以及22个整联蛋白是否直接结合并激活ABL家族激酶。特定的目标2解决了利什曼原虫是否需要HS1信号传导才能进入细胞和感染；它还测试了ABL家族激酶是否激活HS1。在这两个具体目标中，巨噬细胞都将被利什曼原虫感染，以了解对细胞进入的每种信号传导成分的需求，并且小鼠将被利什曼尼亚感染以确定这些蛋白质在疾病中的重要性。还将采用纯化蛋白质和分子生物学技术来破坏蛋白质之间相互作用的生化测定，以阐明此信号传导途径中关键成分之间的接口。通过识别信号传导的整合素，ABL家族激酶和HS1是否允许利什曼尼亚进入细胞并引起小鼠疾病，我们对利什曼原虫感染过程的理解将有所改善。 公共卫生相关性：该项目旨在解释一个称为利什曼原虫的人类寄生虫如何获得细胞的进入，以生存。利什曼尼亚（Leishmania）每年在全球范围内感染数百万人，并杀死其中50,000多人。我们希望，如果我们了解利什曼原虫如何进入细胞，可以开发出新的药物阻断细胞的进入，因此可以由这种寄生虫引起的疾病。