Characterization of Chronic Contusive Spinal Cord Injury and Promotion of Corticospinal Tract Regeneration

慢性挫伤性脊髓损伤的特征及促进皮质脊髓束再生

• 批准号: 10046295
• 负责人: Pengzhe Lu
• 金额: --
• 依托单位: VA SAN DIEGO HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-10-01 至 2022-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10046295
• 关键词: Acute; Adult; Affect; Afghanistan; American; Americas; Animal Model; Animals; Apoptosis; Atrophic; Axon; Back; Bilateral; Bioinformatics; Candidate Disease Gene; Caring; Cervical; Cervical spinal cord structure; Chronic; Cicatrix; Contusions; Corticospinal Tracts; Data; Data Set; Development; Disease; Down-Regulation; Exhibits; Expeditions; Extracellular Matrix; Feeling; Fishes; Functional disorder; Future; Generations; Genes; Genetic Transcription; Grant; Growth; Hand functions; Histologic; Human; In Vitro; Injury; Knowledge; Lesion; Lesion by Stage; Messenger RNA; Methods; Microglia; Mission; Modeling; Molecular; Motor; Movement; Natural regeneration; Neurites; Neuroglia; Neurons; Outcome; Paralysed; Pathway interactions; Patient Care; Process; Quadriplegia; Quality of life; Rattus; Recovery; Refractory; Research; RiboTag; Signal Transduction; Spinal Cord; Spinal Cord Contusions; Spinal Ganglia; Spinal cord injury; Spinal cord injury patients; Stem cell transplant; System; Techniques; Testing; Time; Tissues; Veterans; War; Work; axon growth; axon injury; axon regeneration; base; clinically relevant; experience; guided inquiry; in vivo evaluation; injured; loss of function; molecular pathology; molecular targeted therapies; motor axon regeneration; motor control; nerve stem cell; neurite growth; neuronal cell body; novel therapeutics; postnatal; success; therapeutic development; therapy development; tool; transcriptome; transcriptome sequencing; vector

Characterization of Chronic Contusive Spinal Cord Injury and Promotion of Corticospinal Tract Regeneration Summary Chronic spinal cord injury (SCI) is a post-injury stage when the injury is stable with little additional change. Although the vast majority of SCI are chronic, there are few studies to characterize the chronic injury at histological, cellular, and molecular level in clinical relevant animal models. There are even fewer studies for development of therapeutic treatment of chronic SCI, especially for promotion of supraspinal motor axonal regeneration, including the corticospinal tract (CST) system that is the most important voluntary motor control system in humans. Therefore, we propose to use a chronic clinical relevant moderate bilateral lower cervical contusion model for characterization it at cellular and molecular levels and development of new therapies for improvement of skilled hand function. There are three Specific Aims in this proposal. Aim 1 is to identify cellular mechanisms associated with chronic contusive SCI, to guide discovery of new molecular mechanisms to promote regeneration. We will develop a moderate chronic bilateral lower (C6) cervical contusion model since the majority of SCI patients (65%) are quadriplegic, with loss of hand function that is critical for independence and quality of life. The injured rats survive for up to 12 months to a chronic stage of SCI for characterization of glial scar, extra cellular molecules (ECM) and axonal state. Aim 2 is to identify transcriptional mechanisms associated with chronic contusive SCI, to guide discovery of new molecular mechanisms to promote regeneration. We will use a newly developed technique developed in our lab, Cre-dependent Ribotag vectors, to isolate mRNA specific from chronically injured CST neurons for characterization of their transcriptome using RNAseq. We then compare this transcriptome to intact and sub-acutely injured CST neurons (we already have these datasets) and chronically injured CST neurons that regenerate into neural progenitor cell (NPC) graft to identify key molecular pathways related to cellular/axonal growth that are disrupted, We can therapeutically target these molecular pathways in Aim 3 and future studies. Aim 3 is to test candidate mechanisms to promote recovery of chronically injured CST neurons and promote axonal regeneration after chronic SCI. We will first perform in vitro screen for the potential candidate targets identified in Aim2 to promote neurite growth of postnatal cortical neurons and adult dorsal root ganglion (DRG) neurons. We then test these candidate genes to promote regeneration of chronically injured CST in combination with NPC transplants that serve as a permissive cellular substrate. Findings of this work will substantially extend our knowledge of chronic SCI and develop potential treatment for chronic SCI.

慢性压迫性脊髓损伤的特点及促进治疗 皮质脊髓束再生 摘要 慢性脊髓损伤(Sci)是损伤后的一个阶段，此时损伤是稳定的，几乎没有额外的损伤。 变化。尽管绝大多数脊髓损伤是慢性的，但很少有研究来描述慢性脊髓损伤的特征。 临床相关动物模型的组织学、细胞和分子水平的损伤。甚至还有 慢性脊髓损伤治疗发展的研究较少，尤其是促进 脊髓上运动性轴突再生，包括皮质脊髓束(CST)系统 人类重要的自主运动控制系统。因此，我们建议使用慢性临床 相应的双侧下颈椎中度挫伤模型的细胞学和分子生物学特征 改善熟练手功能的新疗法的水平和发展。 这项提议有三个具体目标。目标1是确定相关的细胞机制 指导发现新的分子机制促进慢性挫伤性脊髓损伤 再生。我们将建立一种中度慢性双侧下颈椎(C6)挫伤模型 大多数脊髓损伤患者(65%)为四肢瘫痪，手功能丧失对 独立性和生活质量。受伤的大鼠存活长达12个月，直到慢性脊髓损伤阶段 用于表征胶质瘢痕、细胞外分子(ECM)和轴突状态。目标2是确定 与慢性挫伤性脊髓损伤相关的转录机制，以指导新的发现 促进再生的分子机制。我们将使用一种新开发的技术 我们实验室开发的依赖Cre的核标签载体，用于从慢性损伤中分离特异性mRNA 利用RNAseq鉴定CST神经元的转录组。然后我们比较这一点 转录组到完整的和亚急性损伤的CST神经元(我们已经有这些数据集)和 慢性损伤的CST神经元再生为神经前体细胞(NPC)以识别关键 与细胞/轴突生长相关的分子通路被破坏，我们可以治疗靶点 目标3中的这些分子途径以及未来的研究。目标3是测试候选机制，以 促进慢性损伤的CST神经元的恢复和促进轴突再生 慢性脊髓损伤。我们将首先对AIM2中确定的潜在候选靶点进行体外筛选 促进出生后皮质神经元和成年背根节(DRG)神经元的轴突生长。我们 然后测试这些候选基因，以促进慢性损伤的CST的再生 鼻咽癌移植作为一种允许的细胞底物。这项工作的发现将极大地 扩大我们对慢性脊髓损伤的认识，开发治疗慢性脊髓损伤的潜在方法。