Role of Lyn in SLE-like autoimmune disease in mice

Lyn 在小鼠 SLE 样自身免疫性疾病中的作用

• 批准号: 6936050
• 负责人: Neetu Gupta
• 金额: $ 11.02万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-15 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6936050
• 关键词: B cell receptor; B lymphocyte; active sites; autoimmune disorder; biological signal transduction; gene targeting; genetically modified animals; green fluorescent proteins; immunoglobulin genes; kidney disorder; laboratory mouse; lupus nephritis; protein engineering; protein localization; protein structure function; protein tyrosine kinase; receptor expression; systemic lupus erythematosus; western blottings

DESCRIPTION (provided by applicant): Lyn is the predominant Src family kinase in B cells and has a unique role in the feedback regulation of BCR signaling. Lyn knockout mice have hyperactive B cells and a phenotype resembling the human condition, systemic lupus erythematosus (SLE) with severe kidney disease. The goals of the proposed research are to identify the physical site of inhibitory signaling mediated by Lyn in B cells, and to use gene therapy to ameliorate the autoimmune kidney disease in Lyn-deficient mice. Lyn will be targeted to the lipid raft or non-raft compartment of the plasma membrane by engineering appropriate chimeric constructs. The chimeric proteins will be tested for their ability to reconstitute signaling by ectopic expression in Lyn-deficient B cells in vitro. The contribution of lipid raft localization of Lyn towards development of autoimmune disease will be tested in vivo by creating knock-in mice expressing raft-restricted or raft-excluded Lyn instead of wild type Lyn. Finally, an attempt will be made to ameliorate the lupus-nephritis phenotype of Lyn-deficient mice by repopulating them with autologous hematopoeitic progenitors reconstituted with wild type Lyn. I have extensive experience in studying immune cell signaling with emphasis on subcellular localization of signaling intermediates during B cell activation. The proposed research will expand my in vitro signaling and cell biology expertise to the study of in vivo mouse models of human autoimmune disease and retrovirus-mediated stem cell transplantation as a means of gene therapy. I will collaborate with investigators within and outside UCSF to execute the research plan. The proposed studies will generate critical molecular and cellular tools to establish a strong research program that aims to study the immune cell defect in SLE-like kidney disease and use gene therapy as a means of intervention. Ultimately, this project will enable me to develop into an independent investigator and pursue a tenure-track faculty position

描述（由申请人提供）： Lyn是B细胞中主要的SRC家族激酶，并且在BCR信号的反馈调节中具有独特的作用。 Lyn基因敲除小鼠具有多动的B细胞和类似于人类状况的表型，具有严重肾脏疾病的全身性红斑狼疮（SLE）。拟议的研究的目标是确定LYN在B细胞中介导的抑制性信号的物理部位，并使用基因治疗来改善Lyn缺陷小鼠中的自身免疫性肾脏疾病。 Lyn将通过工程适当的嵌合构建体来针对质膜的脂质筏或非羊皮室。嵌合蛋白将在体外通过异位表达通过异位表达重新构建信号传导的能力进行测试。 LYN的脂质筏定位​​对自身免疫性疾病发展的贡献将在体内进行测试，从而在体内产生表达Raft限制性或筏的lyn而不是野生型Lyn的敲门型小鼠。最后，将尝试通过用自体造血祖细胞重新培养Lyn缺陷小鼠的狼疮核表型，并用野生型Lyn重构。 我在研究免疫细胞信号传导方面具有丰富的经验，重点是B细胞激活期间信号中间体的亚细胞定位。拟议的研究将把我的体外信号传导和细胞生物学专业知识扩展到人体自身免疫性疾病的体内小鼠模型和逆转录病毒介导的干细胞移植作为基因治疗的一种手段。我将与UCSF内外的调查人员合作执行研究计划。拟议的研究将生成关键的分子和细胞工具，以建立强大的研究计划，旨在研究类似SLE的肾脏疾病中的免疫细胞缺陷，并使用基因治疗作为干预手段。最终，该项目将使我能够发展成为一个独立的调查员并从事终身任职职位