Small Molecule Induced Astrogliogenesis

小分子诱导星形胶质细胞生成

• 批准号: 8078023
• 负责人: Jeffrey D Rothstein
• 金额: $ 24.6万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8078023
• 关键词: Adult; Aftercare; Alzheimer' s Disease; Animals; Astrocytes; Biological Assay; Brain; CSPG4 gene; Cell Line; Cells; Chemicals; Clinic; Disease model; Dose; DsRed; Engineering; Engraftment; Epilepsy; Evaluation; FDA approved; Foundations; Functional disorder; Future; Generations; Glial Fibrillary Acidic Protein; Health; Human; Huntington Disease; In Vitro; Injury; Lead; Libraries; Luciferases; Multiple Sclerosis; Mus; Nerve Degeneration; Neuroglia; Oligodendroglia; Pharmaceutical Preparations; Principal Investigator; Property; Proteins; Reagent; Reporter; Reporting; Rodent; Rodent Model; Screening procedure; Sorting - Cell Movement; Staging; Stem cells; Testing; Therapeutic; Transverse Myelitis; Validation; adult stem cell; base; drug discovery; fetal; follow-up; gliogenesis; immortalized cell; in vitro activity; in vivo; mouse model; nervous system disorder; novel; progenitor; programs; promoter; protein function; research study; response; small molecule; stem cell therapy; tool; transdifferentiation

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

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系统疾病的一个组成部分。替换这些细胞可能是一种新颖且有效的疗法，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体大脑祖细胞，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。我们已经为这些测定产生了合适的报告细胞和动物，包括啮齿动物 NG2-dsRed/GLT1-eGFP、NG2-dsRed-/GLT1 荧光素酶和人神经胶质祖细胞/EAAT2 荧光素酶细胞系，所有这些都适合鉴定将祖细胞分化为星形胶质细胞的分子。我们还生成了必要的小鼠模型，包括 NG2/GLT1 BAC 报告小鼠，以评估新一代星形胶质细胞的体内功效。对于未来的研究，我们将在我们的神经退行性啮齿动物模型（例如 G93A SOD1 小鼠）中测试活性化合物，以确定这些药物在患病小鼠模型中生成新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测定的 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。目标 2) 从啮齿动物报告细胞中的多种化合物库和 FDA 批准的药物库中鉴定星形胶质细胞转化小分子，然后对这些胶质生成药物的新星形胶质细胞生成进行功能验证。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和体外功能活性。 PHS 398/2590（修订版 11/07） 页面延续 格式页面 公共健康相关性：该提案仅关注使用相关啮齿动物和人类祖细胞作为发现工具，以识别能够使体内现有的成年神经胶质祖细胞转分化并导致新神经胶质细胞生成的药物。新星形胶质细胞的产生对于神经系统疾病具有治疗价值，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。最重要的是，它可以产生成体干细胞疗法——通过激活内源性成体中枢神经系统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（修订版 11/07） 页面延续 格式页面