Regulation of Mast Cell Development and Function

肥大细胞发育和功能的调节

基本信息

批准号：
7156984
负责人：
Stephen Joseph Galli
金额：
$ 41.91万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-09-01 至 2011-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7156984
关键词：
1-Phosphatidylinositol 3-Kinase 2,4-Dinitrophenol 5-(6)-carboxyfluorescein diacetate succinimidyl ester Abbreviations Accounting Address Affect Affinity Allergic Anaphylaxis Antibodies Antigens Atopic Dermatitis Attention Bone Marrow Bromodeoxyuridine Cell Degranulation Cell Survival Cell secretion Chromosomes, Human, Pair 10 Chymase Class Coupling Cues Cyclin-Dependent Kinases Dermal Development Disease Economics Effector Cell Elements Endothelin-1 Esters Exhibits Extracellular Signal Regulated Kinases Extrinsic asthma Fetal Liver Glutathione S-Transferase Goals Green Fluorescent Proteins Guanine Nucleotide Exchange Factors Health Systems Agencies Hexosaminidases Histamine Histocytochemistry Homologous Gene Human IgE IgE Receptors Immune response Immunohistochemistry In Vitro Individual Inflammation Inflammatory Interleukin-3 Interleukins Iodine-131 Human Serum Albumin Leukotrienes Lipids MEKs Mediator of activation protein Mitogen-Activated Protein Kinases Molecular Monoclonal Antibodies Mus Mutation Nucleotides Numbers PTEN gene Passive Cutaneous Anaphylaxis Patients Peritoneal Phenotype Phosphotransferases Propidium Diiodide Prostaglandins Prostaglandins D Proteins Proto-Oncogene Protein c-kit Rattus Recombinants Regulation Reporting Rhinitis Role Serine Serum Serum Albumin Signal Pathway Signal Transduction Skin Stem Cell Factor Structure TNF gene Tissues Toll-like receptors Tumor Necrosis Factor-alpha Tumor Necrosis Factors Ubiquitin Ubiquitination Vacuolar Protein Sorting Venous Work Zinc Fingers beta-n-acetylhexosaminidase burden of illness cost cytokine embryonic stem cell human TNF protein in vivo mast cell mutant novel therapeutics phosphatidylinositol 3,4,5-triphosphate receptor response subcutaneous tensin ubiquitin ligase

项目摘要

DESCRIPTION (provided by applicant): Mast cells (MCs) are major effector cells in IgE antibody-dependent allergic disorders, such as anaphylaxis, allergic rhinitis and atopic asthma, which account for a very large burden of illness and economic costs in the developed world. MCs are also critical for the optimal expression of certain innate immune responses. While much attention has been focused on the elements which positively regulate the IgE- and specific antigen (Ag)-dependent secretion of pro-inflammatory mediators and cytokines from MCs, the molecular mechanisms which can suppress the magnitude and/or duration of such responses have been less studied. We recently reported that RabGEF1 (Rab guanine nucleotide exchange factor 1) can negatively regulate Ras-dependent signaling pathways, and the secretion of all three classes of mediators (pre-formed, lipid and cytokine), in MCs stimulated with IgE and specific Ag. More recently, we found that RabGEF1 also importantly regulates responses in MCs which are elicited by the major survival, developmental and proliferation factor for MCs, SCF (stem cell factor, the c-Kit ligand). Notably, our Rabgef1-/- mice exhibit severe inflammation of the skin associated with increased numbers of MCs, evidence of dermal MC degranulation (i.e., "activation"), and increased levels of histamine and IgE in the serum. The central questions which we now wish to address are: By what molecular mechanisms does RabGEF1 influence MC development, activation and function, and to what extent might these actions of RabGEF1 on MCs account for some of the dramatic phenotypic abnormalities observed in Rabgef1-/- mice? In Aim 1. we will investigate how RabGEF1, and its individual functional domains, can negatively regulate mouse MC activation induced by signaling via FceRI or c-Kit (the SCF receptor) in vitro or in vivo, and will identify and characterize RabGEF1-interacting proteins and their downstream effectors in MCs which have been activated via these receptors. In Aim 2, we will define the mechanisms by which RabGEF1 can regulate the survival, development, phenotype & proliferation of MCs in vitro and in vivo. The in vivo studies will take advantage of our ability to transfer in wfro-derived MCs which lack, or express mutant forms of, RabGEF1 into the tissues of c-kit mutant, KitW/W-v or KitW-sh/W-sh genetically MC-deficient mice (which express wild type RabGEF1). We thus can study the effects of RabGEF1 on MCs in mice in which only the MCs lack, or express mutant forms of, RabGEF1. Elucidating how RabGEF1 negatively regulates FceRI- or c-Kit-dependent signaling in MCs will increase our understanding of the regulation of MC activation and development, which is the long-term goal of this project. Such work also may help in the development of new therapeutic approaches for the alleviation of diseases, such as asthma and atopic dermatitis, which are associated with IgE-dependent MC activation and, in many patients, with increased numbers of MCs in the affected tissues.

描述（由申请人提供）：肥大细胞（MCS）是IgE抗体依赖性过敏性疾病中的主要效应细胞，例如过敏反应，过敏性鼻炎和特应性哮喘，这是发达国家中疾病和经济成本的巨大负担。 MC对于某些先天免疫反应的最佳表达也至关重要。尽管很多关注集中在阳性调节MCS的促炎性介质和细胞因子的依赖性和特异性抗原（Ag）依赖性分泌的元素上，但研究了这种反应的大小和/或持续时间的分子机制的研究较少。我们最近报道说，RabGeF1（Rab Guanine核苷酸交换因子1）可以负调节RAS依赖性信号通路，并且在IgE和特定Ag刺激的MC中，MCS中的MCS中，所有三种介体（预成型，脂质和细胞因子）的分泌。最近，我们发现RABGEF1还重要地调节了MC中的反应，这是由MCS的主要生存，发育和增殖因子SCF（干细胞因子，C-KIT配体）引起的。值得注意的是，我们的Rabgef1 - / - 小鼠表现出与MC数量增加相关的皮肤的严重炎症，皮肤MC脱粒的证据（即“激活”）以及血清中组胺和IgE的水平增加。我们现在希望解决的主要问题是：Rabgef1通过哪些分子机制影响MC的发展，激活和功能，Rabgef1对MCS的这些作用在多大程度上占RABGEF1 - / - 小鼠中某些戏剧性表型异常的影响？在AIM 1中。我们将研究RabGeF1及其单个功能结构域如何通过FCERI或C-KIT或C-KIT（SCF受体）在体外或体内负调节小鼠MC激活，并将识别和表征RabGEF1交互的蛋白质及其在通过这些受体激活的MC中的RabGEF1交互蛋白及其下游效应器。在AIM 2中，我们将定义Rabgef1可以调节MC在体外和体内的生存，发育，表型和增殖的机制。体内研究将利用我们在缺乏或表达rabgef1突变形式的WFRO衍生的MC中转移的能力，将Rabgef1（rabgef1）纳入c-kit突变体的组织，kitw/w-v或kitw-sh/w-sh/w-sh/w-sh/w-sh遗传上有mc缺乏的小鼠（表达野生型rabgef1）。因此，我们可以研究Rabgef1对只有MC缺乏或表达突变形式的Rabgef1的MC的影响。阐明RABGEF1在MC中如何负面调节FCERISI依赖性信号将增加我们对MC激活和开发调节的理解，这是该项目的长期目标。这项工作还可能有助于开发新的治疗方法，以减轻与IGE依赖性MC激活有关的哮喘和特应性皮炎，例如哮喘和特应性皮炎，并且在许多患者中，受影响组织中的MC数量增加。