Evaluation of Novel Axon Regeneration Targets for Spinal Cord injury Therapy

脊髓损伤治疗新轴突再生靶点的评价

• 批准号: 9196896
• 负责人: MARC HAMMARLUND
• 金额: $ 38.27万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9196896
• 关键词: Adult; Affect; Alleles; Animal Model; Axon; Axotomy; Bioavailable; Biological Assay; Biological Neural Networks; Caenorhabditis elegans; Cell Culture Techniques; Cell Transplantation; Cell membrane; Cells; Cellular biology; Chemicals; Chest; Control Groups; Contusions; Coupled; Data; Development; Dorsal; Enzymes; Equilibrium; Evaluation; Exocytosis; Family member; Future; GTP Binding; Gene Deletion; Genes; Genetic Screening; Goals; Growth; Growth Cones; Guanosine Triphosphate Phosphohydrolases; Human; INPPL1 gene; Immunoprecipitation; In Vitro; Knockout Mice; Lasers; Lipids; Mammals; Membrane; Membrane Protein Traffic; Methods; Modeling; Monitor; Mus; Mutant Strains Mice; Natural regeneration; Nervous System Physiology; Nervous System Trauma; Nervous system structure; Neuraxis; Neurologic; Neurons; Organism; Outcome Measure; Pathway interactions; Patients; Peripheral Nervous System; Phenotype; Pre-Clinical Model; Process; Proteins; Recovery; Recovery of Function; Rodent; Role; S-nitro-N-acetylpenicillamine; Signal Transduction; Site; Spinal Cord; Spinal Cord Contusions; Spinal cord injury; Synapses; Synaptic Vesicles; System; Testing; Therapeutic Intervention; Trauma recovery; Traumatic CNS injury; Traumatic injury; Vesicle; Work; axon growth; axon regeneration; base; disability; genome-wide; genome-wide analysis; in vivo; inhibitor/antagonist; knock-down; knockout gene; loss of function; mutant; nervous system disorder; neurogenesis; neurological recovery; neuronal growth; novel; novel therapeutics; prenylation; rab GTP-Binding Proteins; receptor mediated endocytosis; relating to nervous system; repaired; research study; restoration; screening; small hairpin RNA; synaptogenesis; therapeutic development; tool; trafficking; validation studies

SUMMARY The CNS of adult mammals, as compared to the peripheral nervous system of mammals or the nervous system of other organisms, has extremely limited capacity for axonal regeneration. Specific factors limiting adult mammalian regeneration of axons have been identified, but they provide an incomplete explanation for poor adult mammalian CNS regeneration. We have completed a genome-wide shRNA-based screen for endogenous genes limiting the repair of axons in the mammalian CNS. We have also conducted experiments to identify conserved genes that affect axon regeneration in the model organism C. elegans. Factors common to both experimental systems are expected to identify fundamental mechanisms in regeneration that are likely to affect the equivalent process in human patients. We aim to study and develop the translational potential of those evolutionarily conserved mechanisms here. From our studies we have selected one evolutionarily conserved pathway identified both in mouse cell culture and in C. elegans axon regeneration. It is bioinformatically the most enriched gene set in the primary mammalian screen data, with multiple family members identified, and also regulates regeneration in C. elegans. The relevance of the pathway will be tested in preclinical models of traumatic spinal cord injury. Multiple steps in the pathway will be assessed in rodent spinal cord injury models. Both gene deletion strains and pharmacological inhibition will be studied to provide a validated pathway for future therapeutic development. While we will focus on one particular pathway regulating membrane traffic in the axon, we will utilize both laser axotomy and mouse spinal cord traumatic injury to explore additional pathways identified in the primary screen. This project builds on genetic screens in the mature mammalian central nervous system and C. elegans to analyze novel mechanisms that promote axon regeneration after mammalian spinal cord injury. The findings will have high relevance for the development of novel therapeutics for neurological disorders.

摘要 成年哺乳动物的中枢神经系统，与哺乳动物的周围神经系统或神经系统相比 轴突再生能力极其有限。特定的限制因素 成年哺乳动物轴突的再生已被确认，但它们提供了对 成年哺乳动物中枢神经系统再生能力差。我们已经完成了基于shRNA的全基因组筛选 限制哺乳动物中枢神经系统轴突修复的内源性基因。我们还进行了一些实验 在模式生物秀丽线虫中识别影响轴突再生的保守基因。共同因素 这两个实验系统都有望确定再生的基本机制 很可能会影响人类患者的相同过程。我们的目标是研究和开发 这些进化上保守的机制的翻译潜力。 从我们的研究中，我们选择了一条在小鼠细胞培养中发现的进化保守的途径 线虫的轴突再生。这是生物信息学上最丰富的基因集在初级 哺乳动物的筛选数据，识别出多个家庭成员，也调节线虫的再生。 该途径的相关性将在创伤性脊髓损伤的临床前模型中进行测试。多步骤 将在啮齿动物脊髓损伤模型中进行评估。基因缺失菌株和 将对药物抑制进行研究，为未来的治疗开发提供一条有效的途径。 虽然我们将专注于调节轴突中膜交通的一种特殊途径，但我们将利用这两种激光 轴突切断和小鼠脊髓创伤性损伤探索在初级筛选中确定的其他途径。 该项目建立在成熟哺乳动物中枢神经系统和线虫的遗传屏幕上，以 分析哺乳动物脊髓损伤后促进轴突再生的新机制。调查结果 将对神经系统疾病的新疗法的开发具有很高的相关性。