Small Molecule Induced Astrogliogenesis

描述（由申请人提供）：星形胶质细胞损伤和/或功能障碍似乎是许多神经系统疾病的组成部分。这些细胞的替代可能是一种新的和有效的治疗方法-但移植方法在治疗上具有挑战性。成体NG 2细胞-少突胶质细胞和星形胶质细胞的常驻成体脑祖细胞在啮齿动物和人类CNS中丰富。我们建议筛选啮齿动物和人类NG 2细胞与化学/药物库（单独和矩阵组合），以确定能够分化这些细胞在体外和体内星形胶质细胞的小分子。我们已经产生了用于这些测定的合适的报告细胞和动物，包括啮齿动物NG 2-dsRed/GLT 1-eGFP、NG 2-dsRed-/GLT 1荧光素酶和人神经胶质祖细胞/EAAT 2荧光素酶细胞系-所有这些都适合于鉴定将祖细胞分化为星形胶质细胞的分子。我们还产生了必要的小鼠模型，包括NG 2/GLT 1 BAC报告小鼠，以评估新星形胶质细胞生成的体内功效。对于未来的研究，我们将在我们的啮齿动物神经变性模型（例如G93 A SOD 1小鼠）中测试活性化合物，以确定药物在患病小鼠模型中产生新星形胶质细胞的功效。 本研究的目的包括：Aim 1）用于药物发现测定的NG 2-祖细胞/星形胶质细胞报告细胞系的产生和验证，包括来自NG 2-星形胶质细胞报告小鼠的啮齿动物NG 2报告细胞系和人祖细胞-报告细胞系。目的2）从啮齿动物报告细胞中的多种化合物文库和FDA批准的药物文库中鉴定星形胶质细胞转化小分子，然后从这些胶质细胞生成药物中验证新的星形胶质细胞生成的功能。最后，新的胶质细胞生成药物将在人祖细胞系中进行验证，包括细胞特异性标记和体外功能活性。PHS 398/2590（Rev. 11/07） 公共卫生相关性：该提案仅关注使用相关的啮齿动物和人类祖细胞作为发现工具，以鉴定可以在体内转分化现有的成年神经胶质祖细胞并导致产生新神经胶质细胞的药物。新的星形胶质细胞的产生可能对神经系统疾病有治疗价值，包括ALS、亨廷顿病、阿尔茨海默病、癫痫、多发性硬化症和横肌萎缩症。最重要的是，它可以产生成人干细胞疗法-通过激活内源性成人CNS祖细胞分化成新的星形胶质细胞-从而消除对基于外部细胞的治疗的需要。PHS 398/2590（Rev. 11/07）

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