Role of SAP,SLAM and Fyn in NKT Cell Ontogeny and Activation

SAP、SLAM 和 Fyn 在 NKT 细胞个体发育和激活中的作用

• 批准号: 7662291
• 负责人: KIM Erika NICHOLS
• 金额: $ 41.13万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-18 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7662291
• 关键词: Address; Adoptive Cell Transfers; Agonist; Animals; Antibody Formation; Autoimmunity; Binding; Biochemical Pathway; Biological Assay; Biological Models; CD4 Positive T Lymphocytes; Cell Differentiation process; Cell Lineage; Cell physiology; Cells; Complex; Cytoplasmic Tail; Data; Defect; Development; Disease; Galactosylceramides; Genes; Genetic; Germ-Line Mutation; Goals; Human; Human Biology; Immunity; Immunodeficiency and Cancer; Immunologic Deficiency Syndromes; Immunologics; In Vitro; Infection; Infection Control; Inherited; Knock-in Mouse; Ligands; Ligation; Link; Lymphocyte; Lymphocytic choriomeningitis virus; Lymphoma; Malignant Neoplasms; Medical; Methods; Molecular; Monitor; Monoclonal Antibodies; Mouse Strains; Mus; Mutation; Natural Killer Cells; Outcome; Pathogenesis; Pathway interactions; Patients; Pattern; Phenotype; Phosphorylation; Phosphotyrosine; Predisposition; Production; Property; Proto-Oncogene Proteins c-fyn; Rare Diseases; Reaction; Receptor Signaling; Recruitment Activity; Regulation; Research Personnel; Retroviridae; Role; SLAM family receptor; Signal Pathway; Signal Transduction; T-Cell Activation; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Therapeutic; Viral; Virus Diseases; Wild Type Mouse; X-Linked lymphoproliferative disorders; base; crosslink; cytokine; cytotoxicity; domain mapping; human disease; improved; in vivo; insight; killer T cell; killings; novel; pathogen; progenitor; ras GTPase-Activating Proteins; receptor; receptor function; response; restoration; src-Family Kinases; thymocyte; tumor

DESCRIPTION (provided by applicant): Through studies of the inherited immunodeficiency X-linked lymphoproliferative disease (XLP), we discovered that the adaptor molecule SAP is required for development of natural killer T (NKT) cells, a subset of regulatory lymphocytes with essential functions in innate and adaptive immunity. In particular, we showed that XLP is caused by mutations in the SAP/SH2D1A gene and that SAP-deficient humans and mice lack NKT cells. To extend these findings and exploit them for more general therapeutic purposes, we plan to characterize the signaling mechanisms used by SAP to regulate NKT cell ontogeny and function. In T lymphocytes, SAP recruits the Src family tyrosine kinase Fyn to the cytoplasmic domain of the SLAM receptor, thereby promoting downstream phosphotyrosine signals that are required for TH2-type cytokine production. Our preliminary studies suggest that SAP controls NKT cell ontogeny through Fyn, but also through novel Fyn-independent pathways. Based on these observations, we hypothesize that SAP is a critical molecule required for NKT cell development. Furthermore, SAP may promote NKT cell ontogeny via signaling pathways distinct from those used by this adaptor in other cell lineages, such as CD4+ T cells. Because NKT cells have potent immunoregulatory properties, we also propose that the NKT cell defect contributes to the phenotypes of SAP-deficiency, including enhanced susceptibility to infection, humoral insufficiency and development of lymphoma. We will investigate these hypotheses through 3 specific aims. In Aim 1, we will use in vitro culture methods and in vivo mouse studies to explore the biochemical pathways involving SAP that guide NKT cell ontogeny. In Aim 2, we will manipulate SAP expression and function in mature NKT cells to determine how SAP-dependent signals control NKT cell functions, such as cytokine production and cytotoxicity. In Aim 3, we will use adoptive cell transfer to restore NKT cell function in NKT cell-deficient and Sap-/- mice to determine the importance of this lineage during clearance of virus infection and regulation of anti-viral immunity. The medical relevance of NKT cells is established, but the molecular mechanisms controlling their development and function are not well understood. Our analyses of XLP identify SAP as a crucial regulator of NKT cell ontogeny and illustrate how studies of a rare disorder can produce important insights into human biology and disease pathogenesis. By further defining the role of SAP in NKT cells, we hope to identify novel signaling pathways that can be targeted to improve the treatment of XLP as well as more common human diseases, such as immunodeficiency, autoimmunity and cancer.

描述（由申请人提供）：通过对遗传性免疫缺陷性 X 连锁淋巴组织增生性疾病 (XLP) 的研究，我们发现接头分子 SAP 是自然杀伤 T (NKT) 细胞（具有基本功能的调节性淋巴细胞的一个子集）发育所必需的在先天性和适应性免疫中。特别是，我们发现 XLP 是由 SAP/SH2D1A 基因突变引起的，并且 SAP 缺陷的人类和小鼠缺乏 NKT 细胞。为了扩展这些发现并将其用于更普遍的治疗目的，我们计划表征 SAP 用于调节 NKT 细胞个体发育和功能的信号机制。在 T 淋巴细胞中，SAP 将 Src 家族酪氨酸激酶 Fyn 招募到 SLAM 受体的胞质结构域，从而促进 TH2 型细胞因子产生所需的下游磷酸酪氨酸信号。我们的初步研究表明 SAP 通过 Fyn 控制 NKT 细胞个体发育，但也通过新颖的 Fyn 独立途径控制 NKT 细胞个体发育。基于这些观察，我们假设 SAP 是 NKT 细胞发育所需的关键分子。此外，SAP 可能通过不同于该接头在其他细胞谱系（例如 CD4+ T 细胞）中使用的信号通路来促进 NKT 细胞个体发育。由于 NKT 细胞具有有效的免疫调节特性，我们还提出 NKT 细胞缺陷会导致 SAP 缺陷的表型，包括对感染的易感性增强、体液不足和淋巴瘤的发展。我们将通过 3 个具体目标来研究这些假设。在目标 1 中，我们将利用体外培养方法和体内小鼠研究来探索涉及 SAP 指导 NKT 细胞个体发育的生化途径。在目标 2 中，我们将操纵成熟 NKT 细胞中的 SAP 表达和功能，以确定 SAP 依赖性信号如何控制 NKT 细胞功能，例如细胞因子的产生和细胞毒性。在目标 3 中，我们将使用过继细胞移植来恢复 NKT 细胞缺陷和 Sap-/- 小鼠的 NKT 细胞功能，以确定该谱系在清除病毒感染和调节抗病毒免疫过程中的重要性。 NKT 细胞的医学意义已确立，但控制其发育和功能的分子机制尚不清楚。我们对 XLP 的分析确定 SAP 是 NKT 细胞个体发育的关键调节因子，并说明了对罕见疾病的研究如何能够对人类生物学和疾病发病机制产生重要的见解。通过进一步明确 SAP 在 NKT 细胞中的作用，我们希望找到新的信号传导途径，以改善 XLP 以及更常见的人类疾病（如免疫缺陷、自身免疫和癌症）的治疗。