Novel Factors Promoting Retinal Ganglion Cell Growth

促进视网膜神经节细胞生长的新因素

• 批准号: 7026572
• 负责人: Donald J. Zack
• 金额: $ 20.43万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7026572
• 关键词: axon; dendrites; library; optic nerve; retinal ganglion; small molecule

DESCRIPTION (provided by applicant): The NIH Roadmap emphasizes how the discovery of bioactive "small molecules" can help both in the exploration of "cellular functions at the molecular level" and the development of new drugs. Reflecting the importance of such molecules, one of the Molecular Libraries Roadmap's Initiatives is to support the construction and screening of small molecule libraries. Small molecules and peptides support retinal ganglion cell (RGC) survival and neurite outgrowth by modulating various cellular signaling pathways. Identification of additional such molecules is the focus of this grant. Forskolin and BDNF are examples of molecules that that were originally found to support RGCs in culture and later demonstrated to have efficacy in animal models of optic nerve injury. Identification of novel small molecules that support RGCs in cell culture offers two potential benefits. Such molecules will make possible new lines of inquiry into RGC function at the molecular level. Secondly, factors supporting RGCs in cell culture are a logical source of candidates that may have translational, therapeutic benefit in preserving or restoring vision in patients with blinding optic nerve diseases such as glaucoma, optic nerve stroke, and optic neuritis. Small molecules have potential advantages compared to larger molecules, such as proteins, in that they may be more readily developed into drugs and may be more easily delivered to the eye. A rational and comprehensive way to identify functionally interesting small molecules is to combine a sensitive bioassay with a high throughput screen. We have successfully initiated such a screen, assaying cultures of immunopurified rat RGCs with an automated fluorescence-based image analysis system that measures cell survival and neurite outgrowth. The objective of this proposal is to expand this screen so as to identify, validate, and optimize novel pharmaceutical compounds that promote survival and regeneration of RGCs. More specifically, this grant will support the continued survey of a "small molecule" library for RGC survival and neurite outgrowth factors, confirm the bioactivity of those hits by demonstrating a dose response relationship, and further determine the essential element that exhibits the bioactivity. Reagents identified in the screen will be the subject of future hypothesis driven research to characterize their mechanism of action in vitro and further explore their therapeutic potential in vivo.

描述（由申请人提供）：NIH路线图强调了生物活性“小分子”的发现如何有助于探索“分子水平的细胞功能”和新药开发。为了反映这些分子的重要性，分子库路线图的倡议之一是支持小分子库的构建和筛选。小分子和肽通过调节各种细胞信号传导途径来支持视网膜神经节细胞（RGC）存活和神经突生长。鉴定更多这样的分子是这项资助的重点。毛喉素和BDNF是最初发现在培养中支持RGCs的分子的例子，后来证明在视神经损伤的动物模型中具有功效。在细胞培养中鉴定支持RGC的新型小分子提供了两个潜在的益处。这些分子将使在分子水平上探究RGC功能的新途径成为可能。其次，细胞培养中支持RGC的因子是候选物的逻辑来源，其可能在患有致盲视神经疾病（如青光眼、视神经中风和视神经炎）的患者中具有保留或恢复视力的转化治疗益处。与较大的分子（如蛋白质）相比，小分子具有潜在的优势，因为它们可以更容易地开发成药物，并且可以更容易地递送到眼睛。鉴定功能感兴趣的小分子的合理和全面的方法是将灵敏的生物测定与高通量筛选结合联合收割机。我们已经成功地启动了这样一个屏幕，测定培养免疫纯化的大鼠视网膜神经节细胞与自动荧光为基础的图像分析系统，测量细胞存活和神经突生长。该提案的目的是扩大该筛选，以鉴定、验证和优化促进RGC存活和再生的新型药物化合物。更具体地说，这笔赠款将支持对RGC存活和神经突生长因子的“小分子”文库的持续调查，通过证明剂量反应关系来确认这些命中的生物活性，并进一步确定表现出生物活性的基本元素。筛选中确定的试剂将成为未来假设驱动研究的主题，以表征其体外作用机制，并进一步探索其体内治疗潜力。