Small Molecule Induced Astrogliogenesis

描述(由申请人提供)：星形胶质细胞的损伤和/或功能障碍似乎是许多神经疾病的一个组成部分。这些细胞的替换可能是一种新颖而有效的治疗方法--但植入方法在治疗上具有挑战性。成体NG2细胞--少突胶质细胞和星形胶质细胞的常驻成人脑前体细胞在啮齿动物和人类中枢神经系统中大量存在。我们建议使用化学/药物文库(单独和基质组合)来筛选啮齿动物和人类NG2细胞，以鉴定能够在体外和体内将这些细胞分化为星形胶质细胞的小分子。我们已经为这些检测建立了合适的报告细胞和动物，包括啮齿动物NG2-dsRed/GLT1-EGFP、NG2-dsRed-/GLT1荧光素酶和人类神经胶质前体/EAAT2荧光素酶细胞系-所有这些都适合于识别将祖细胞分化为星形胶质细胞的分子。我们还建立了必要的小鼠模型，包括NG2/GLT1 BAC报告小鼠，以评估新生星形胶质细胞生成的体内效果。在未来的研究中，我们将在我们的神经退行性变的啮齿动物模型(例如G93A SOD1小鼠)中测试活性化合物，以确定这些药物在疾病小鼠模型中生成新星形胶质细胞的功效。本研究的目的包括：1)建立和验证NG2前体细胞/星形胶质细胞报告细胞系，包括来自NG2星形胶质细胞报告鼠的啮齿动物NG2报告细胞系和人类祖细胞报告细胞系。目的2)在啮齿动物报告细胞中从多种化合物文库和FDA批准的药物文库中鉴定星形胶质细胞转化小分子，然后对这些胶质形成药物的新的星形胶质形成进行功能验证。最后，新的胶质生成药物将在人类祖细胞系中进行验证，包括细胞特异性标记物和体外功能活性。PHS 398/2590(11/07版)页面续格式页面 公共卫生相关性：这项提案仅专注于使用相关的啮齿动物和人类祖细胞作为一种发现工具，以确定是否有药物可以在体内转分化现有的成年神经胶质前体细胞并导致新的神经胶质细胞的产生。新的星形胶质细胞的产生可能对包括肌萎缩侧索硬化症、亨廷顿病、阿尔茨海默病、癫痫、多发性硬化症和横贯性脊髓炎在内的神经系统疾病具有治疗价值。最重要的是，它可以产生成人干细胞疗法--通过激活内源性成年中枢神经系统祖细胞分化为新的星形胶质细胞--从而消除对外部基于细胞的治疗的需要。PHS 398/2590(11/07版)页面续格式页面

DESCRIPTION (provided by applicant): Injury and/or dysfunction of astroglia appears to be a component of numerous neurological disorders. Replacement of these cells might be a novel and potent therapy- but engraftment approaches are therapeutically challenging. Adult NG2 cells- the resident adult brain progenitors for oligodendroglia and astroglia are abundant in the rodent and human CNS. We propose to screen rodent and human NG2 cells with a chemical/drug library (singly and in matrix combination) to identify small molecules capable of differentiating these cells to astroglia in vitro and in vivo. We have generated appropriate reporter cells and animals for these assays including rodent NG2-dsRed/GLT1-eGFP, NG2-dsRed-/GLT1 luciferase and human glial progenitor/EAAT2 luciferase cell lines-all suitable for identifying molecules that differentiate progenitors into astroglia. We also have generated the necessary mouse models including NG2/GLT1 BAC reporter mice to evaluate the in vivo efficacy of new astrocyte generation. For future studies, we will test active compounds in our rodent models of neurodegeneration (e.g. G93A SOD1 mouse) to determine the efficacy of the agents in generating new astroglial in diseased mouse models. The aims of the study include: Aim1) Generation and validation of NG2-progenitor/astroglial reporter cell lines for use in drug discovery assays, including rodent NG2 reporter cell lines from NG2-astroglia reporter mice and human progenitor-reporter cell lines. Aim 2) Identification of astroglial transforming small molecules from a diverse chemical compound library and a library of FDA approved drugs in rodent reporter cells, followed by functional validation of new astrogliogenesis from these gliogenic drugs. Finally, the new gliogenic drugs will be validated in the human progenitor cell lines including cell-specific markers and functional activity in vitro. PHS 398/2590 (Rev. 11/07) Page Continuation Format Page PUBLIC HEALTH RELEVANCE: This proposal is solely focused on the use of relevant rodent and human progenitor cells as a discovery tool to identify drugs would could transdifferentiate -in vivo-existing adult glial progenitor cells and lead to the generation of new glial cells. The generation of new astroglial cells could be therapeutically valuable for neurological diseases including ALS, Huntington's disease, Alzheimer's disease ,epilepsy, multiple sclerosis and transverse myelitis. Most importantly, it could generate adult stem cell therapy- by activating endogenous adult CNS progenitor cells to differentiate into new astroglia-thereby eliminating need for external cellular based therapy. PHS 398/2590 (Rev. 11/07) Page Continuation Format Page