Development of a conditional T1MP-1 knockout mouse

条件性T1MP-1基因敲除小鼠的研制

基本信息

批准号：
8283645
负责人：
Stephen J Crocker
金额：
$ 7.7万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8283645
关键词：
Apoptotic Area Astrocytes Bacterial Infections Birth Weight Brain Brain Injuries Cells Central Nervous System Diseases Confounding Factors (Epidemiology)Demyelinations Development Disease Disease model Enzymes Exons Extracellular Protein Family Functional RNA Gene Expression Generations Genes Genetic Recombination Glial Fibrillary Acidic Protein Homeostasis Immune Immunology Inflammation Inflammatory Injury Interleukins Introns Knock-out Knockout Mice Litter Size Malignant Neoplasms Matrix Metalloproteinases Mediating Messenger RNA Metalloproteinase Gene Microglia Mouse Strains Mus Neurons Neurosciences Oligodendroglia Physiological Play Procedures Production Proteins Reagent Recovery Regulation Reporting Research Research Project Grants Role Site Source Spinal Cord Synaptic plasticity T-Lymphocyte Technology Testing Tissue Inhibitor of Metalloproteinase-1 Tissues Transgenic Mice Validation Weight Gain Work X Chromosome anticancer research behavior influence brain tissue cell type central nervous system injury cytokine design extracellular macrophage nervous system disorder neuropathology neuroprotection promoter recombinase response tumorigenesis vector virology

项目摘要

DESCRIPTION (provided by applicant): Tissue inhibitor of metalloproteinases-1 (TIMP-1) is a pleiotrophic extracellular protein that is dramatically upregulated in the brain following inflammation or injury. TIMP-1 has a well described function as an endogenous regulator of the proteolytic activities of the matrix metalloproteinases (MMPs) a family of extracellular enzymes that play critical roles in cellular homeostasis, adaptation and tissue remodeling. TIMP-1 also has noted MMP-independent functions including antiapoptotic activity and trophic actions. These latter functions of TIMP-1 have been implicated in synaptic plasticity, neuroprotection, oncogenesis and oligodendrocyte differentiation. TIMP-1 can be expressed by a variety of cell types in the CNS: astrocytes robustly express TIMP-1 in response to inflammatory demyelination or bacterial infection in the CNS, and neurons have been reported to express TIMP-1 following excitotoxic injury. In the course of CNS injury and inflammation, it is also important to point out that TIMP-1 is also expressed by immune cells, including T cells, macrophages and microglia. Thus, TIMP-1 can be rapidly expressed by a variety of cell types offering a myriad of potential roles for TIMP-1 in response to CNS injury or disease. Currently, only global TIMP-1KO mice are available for study meaning that the cellular source of TIMP-1 remains a confounding variable to our understanding on the specific mechanistic roles of TIMP-1 in the brain. To test whether production of TIMP-1 from specific cell types differentially impact neuropathology, we will develop an essential new reagent: a conditional TIMP-1 knockout mouse (TIMP-1cKO) line. This mouse line will significantly advance our understanding of TIMP-1 and its functions in the CNS by enabling us to directly test whether cell specific-TIMP-1 expression impacts the innate potential for brain recovery following injury. The first aim of this proposal will be to develop the new TIMP-1cKO mouse, which will then be crossed with the GFAP-Cre transgenic mouse to specifically knockout TIMP-1 expression from astrocytes throughout the CNS. Astrocyte-specific TIMP-1cKO mice will then be tested and compared with global. Together this work will, (a) provide a valuable new reagent for advanced study on TIMP-1 in a wide variety of disease models, including neurological diseases, and, (b) experimental results from this work will validate the TIMP-1cKO mouse as a viable new mouse line for study of cell-specific deletion of TIMP-1. PUBLIC HEALTH RELEVANCE: Tissue inhibitor of metalloproteinase-1 (TIMP-1) is a protein that can protect the brain from injury and disease in a variety of ways. The purpose of this research project is to develop a new mouse strain where this protein can be eliminated from specific cells, and in so doing allow us to better understand how TIMP-1 can influence the behaviour of different types of cells during disease.

描述（由申请人提供）：金属蛋白酶组织抑制剂-1（TIMP-1）是一种多效细胞外蛋白，在炎症或损伤后大脑中的表达显着上调。 TIMP-1 具有作为基质金属蛋白酶 (MMP) 蛋白水解活性的内源性调节剂的功能，该酶是细胞外酶家族，在细胞稳态、适应和组织重塑中发挥着关键作用。 TIMP-1 还具有独立于 MMP 的功能，包括抗凋亡活性和营养作用。 TIMP-1 的后一功能与突触可塑性、神经保护、肿瘤发生和少突胶质细胞分化有关。 TIMP-1 可由中枢神经系统中的多种细胞类型表达：星形胶质细胞在中枢神经系统炎症性脱髓鞘或细菌感染的反应中强烈表达 TIMP-1，据报道神经元在兴奋性毒性损伤后表达 TIMP-1。在中枢神经系统损伤和炎症过程中，还需要指出的是，免疫细胞也表达TIMP-1，包括T细胞、巨噬细胞和小胶质细胞。因此，TIMP-1 可以由多种细胞类型快速表达，为 TIMP-1 响应 CNS 损伤或疾病提供多种潜在作用。目前，只有全球 TIMP-1KO 小鼠可用于研究，这意味着 TIMP-1 的细胞来源仍然是我们理解 TIMP-1 在大脑中的具体机制作用的一个混杂变量。测试特定细胞类型产生的 TIMP-1 是否会产生不同的影响在神经病理学方面，我们将开发一种重要的新试剂：条件性 TIMP-1 基因敲除小鼠 (TIMP-1cKO) 系。该小鼠系将使我们能够直接测试细胞特异性 TIMP-1 表达是否影响损伤后大脑恢复的先天潜力，从而显着增进我们对 TIMP-1 及其在中枢神经系统中功能的理解。该提案的首要目标是开发新的 TIMP-1cKO 小鼠，然后将其与 GFAP-Cre 转基因小鼠杂交，以特异性敲除整个中枢神经系统星形胶质细胞的 TIMP-1 表达。然后将对星形胶质细胞特异性 TIMP-1cKO 小鼠进行测试并与全球进行比较。这项工作将共同：(a) 为多种疾病模型（包括神经系统疾病）中 TIMP-1 的高级研究提供一种有价值的新试剂，(b) 这项工作的实验结果将验证 TIMP-1cKO 小鼠用于研究细胞特异性删除 TIMP-1 的可行新小鼠系。公共健康相关性：金属蛋白酶-1 组织抑制剂 (TIMP-1) 是一种蛋白质，可以通过多种方式保护大脑免受损伤和疾病。该研究项目的目的是开发一种新的小鼠品系，其中这种蛋白质可以从特定细胞中消除，从而使我们能够更好地了解 TIMP-1 在疾病期间如何影响不同类型细胞的行为。