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一个组成部分。替换这些细胞可能是一种新颖而有效的治疗方法，但移植方法在治疗上具有挑战性。成人NG2细胞是少突胶质细胞和星形胶质细胞的常驻成人脑祖细胞，在啮齿动物和人类中枢神经系统中含量丰富。我们建议使用化学/药物文库（单独和基质组合）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。我们已经为这些实验生成了合适的报告细胞和动物，包括啮齿动物NG2-dsRed/GLT1- egfp， NG2-dsRed-/GLT1荧光素酶和人类胶质祖细胞/EAAT2荧光素酶细胞系，这些细胞系都适合鉴定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，以评估新星形胶质细胞生成的体内功效。在未来的研究中，我们将在我们的啮齿动物神经变性模型（例如G93A SOD1小鼠）中测试活性化合物，以确定这些药物在患病小鼠模型中产生新的星形胶质细胞的功效。该研究的目的包括：目的1)生成和验证用于药物发现试验的NG2祖细胞/星形胶质细胞报告细胞系，包括来自NG2星形胶质细胞报告小鼠的啮齿动物NG2报告细胞系和人类祖细胞报告细胞系。目的2)从多种化合物文库和FDA批准的药物文库中鉴定啮齿动物报告细胞中的星形胶质转化小分子，然后对这些胶质生成药物的新星形胶质生成进行功能验证。最后，将在人类祖细胞系中验证新的胶质原性药物，包括细胞特异性标记和体外功能活性。小灵通398/2590 （Rev. 11/07）页延续格式页

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