Mid-sized GDNF Mimics For Neural Regeneration

中型 GDNF 模拟神经再生

• 批准号: 10811356
• 负责人: KEVIN BURGESS
• 金额: $ 46.47万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10811356
• 关键词: 3-Dimensional; Afferent Neurons; Affinity; Agonist; Axon; Binding; Biological Assay; Biomedical Engineering; Cell Adhesion Molecules; Cell model; Cells; Cellular Assay; Characteristics; Clinical; Clinical Trials; Communication; Complex; Computing Methodologies; Coupled; Coupling; Cues; Dangerousness; Development; Docking; Engineering; Event; Family; Glycosylphosphatidylinositols; Growth; Growth Factor; Hot Spot; Hyaluronic Acid; In Vitro; Investigation; Lead; Lecithin; Libraries; Ligand Binding; Ligands; Lipase; Literature; Maleimides; Mediating; Medicine; Membrane Microdomains; Modeling; Molecular Conformation; Motor Neurons; Nerve Regeneration; Nervous System; Nervous System Trauma; Neural Cell Adhesion Molecules; Neurites; Neurons; Nuclear; Peripheral Nervous System; Pharmaceutical Preparations; Phase III Clinical Trials; Property; Proteins; Recovery; Reporting; Role; Sampling; Series; Serum; Signal Induction; Signal Pathway; Signal Transduction; Spinal Ganglia; Spinal cord injury; Surface; Synapses; Testing; Time; Tissues; Transfection; Translating; Trauma; Traumatic injury; Work; axon growth; axon guidance; clinical trials in animals; crosslink; cytokine; design; experience; experimental study; extracellular; gene therapy; glial cell-line derived neurotrophic factor; improved; in vitro Model; in vivo; innovation; neuronal survival; neurotensin mimic 2; novel; predictive tools; prevent; professor; prototype; receptor; repair model; repaired; response; small molecule; stem cells; success; three-dimensional modeling; two-dimensional

Project Summary Repair of traumatic injuries relies upon glial cell line-derived neurotrophic factor (GDNF), and related extracellular cytokines collectively called GDNF family ligands (GFLs). GFLs interact with solubilized forms of the GDNF-family receptors (sGFRα1–4) forming complexes which then can bind and activate NCAM (nuclear cell adhesion molecule) and RET (REarranged on Transfection) receptors leading to intracellular signaling and a range of responses conducive to neuronal connectivity. GFLs have been tested in animals and in clinical trials. However, they have poor in vivo stabilities, unfavorable tissue permeation characteristics, and are expensive to prepare with batch-to-batch reproduciblity. Gene therapy approaches have also been attempted, but these are extremely risky because continued expression leads to uncontrollable growth post therapy. Few small molecule mimics of GFL•GFRα interface regions have been reported in the literature. This is surprising because appropriate small molecules could cause conformational changes in sGFRαs transforming them into NCAM/RET agonists which may communicate between cells (trans-signaling) to trigger valuable responses for repair of the peripheral nervous system after trauma. Preliminary studies feature design, synthesis, and testing of two mimics of the GDNF loop which is responsible for most if the GFL•GFRα interface interaction (ie the interface “hot loop”). These loop mimics bind GFRα1 (best so far Kd 240 nM), and are currently being tested in cellular models for repair of traumatic injuries to the peripheral nervous system (PNS). This application is to optimize these initial leads and test them more extensively. Year 1 will focus on on design, syntheses, and GFRα1-binding affinities for similar “cyclo-organopeptide hot loop mimics” by the PI (10 – 20 compounds). Free loop mimics with superior GFRα binding affinities, and samples of ones covalently anchored to hyaluronic acid supports (which mimic the media around synapses), will be selected for Aim 2. The PI is an expert on design and synthesis of growth factor hot loop mimics; he will oversee that part of the work closely. In year 2 the emphasis will shift to testing the best hot-loop mimics identified at that time in 2D and 3D-cellular models for PNS recovery from traumatic injury. Active compounds will also be assayed to test if they cause intracellular activation of NCAM and/or RET. That work will be overseen by Professor Sakiyama, the subcontractor on this application, who has extensive experience with GFLs and supported GFLs, particularly GDNF, tested 2D and 3D cellular assays for neurite outgrowth on sensory and motor neurons. She is an expert in neuronal repair.

项目概要 创伤性损伤的修复依赖于神经胶质细胞源性神经营养因子（GDNF）和相关的 细胞外细胞因子统称为 GDNF 家族配体 (GFL) 与溶解的细胞因子相互作用。 GDNF 家族受体 (sGFRα1-4) 的形式形成复合物，然后可以结合并 激活 NCAM（核细胞粘附分子）和 RET（转染重排）受体 导致细胞内信号传导和一系列有利于神经连接的反应。 GFL 已在动物和临床试验中进行了测试，但其体内稳定性较差。 不利的组织渗透特性，并且批次间的制备成本昂贵 还尝试过基因治疗方法，但这些方法风险极大。 因为持续表达会导致治疗后无法控制的生长。 文献中已经报道了 GFL·GFRα 界面区域的模拟物，这是令人惊讶的。 因为适当的小分子可以引起 sGFRα 转化中的构象变化 将它们转化为 NCAM/RET 激动剂，这些激动剂可以在细胞之间进行通讯（转信号）以触发 对于创伤后周围神经系统的修复有价值的反应。 初步研究的特点是设计、合成和测试 GDNF 环的两种模拟物， 负责大部分 GFL•GFRα 界面交互（即界面“热循环”）。 模拟物结合 GFRα1（迄今为止最佳 Kd 240 nM），目前正在细胞模型中进行测试 修复周围神经系统（PNS）的创伤。 该应用程序旨在优化这些初始线索并更具体地测试它们，第一年将重点关注它们。 关于类似“环有机肽热环模拟物”的设计、合成和 GFRα1 结合亲和力 由 PI 提供（10 – 20 种具有卓越 GFRα 结合亲和力的自由环模拟物和样品）。 共价锚定在透明质酸支持物上（模拟突触周围的介质）， 将被选为目标2。PI是生长因子热循环设计和合成方面的专家 模仿；他将密切监督这部分工作，第二年的重点将转向测试。 当时在 2D 和 3D 细胞模型中发现了最佳的热环模拟，用于 PNS 恢复 还将检测活性化合物是否引起细胞内激活。 NCAM 和/或 RET 的工作将由该项目的分包商 Sakiyama 教授负责监督。 应用程序，在 GFL 和支持的 GFL（特别是 GDNF）方面拥有丰富的经验，并经过测试 感觉和运动神经元上神经突生长的 2D 和 3D 细胞测定她是专家。 神经元修复。