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: 10292951
• 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/10292951
• 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.

慢性旋转脊髓损伤和促进的表征 皮质脊髓区域再生 概括 慢性脊髓损伤（SCI）是伤害后损伤稳定的阶段 改变。尽管绝大多数SCI都是慢性的，但很少有研究表征慢性 在临床相关动物模型中，组织学，细胞和分子水平的损伤。甚至有 慢性SCI治疗治疗的开发研究较少，尤其是为了促进 脊柱上轴突上轴突再生，包括最多的皮质脊髓区（CST）系统 人类重要的自愿运动控制系统。因此，我们建议使用慢性临床 相关的中度双侧下颈挫伤模型，以表征在细胞和分子处 新疗法的水平和开发，以改善熟练的手部功能。 该提案中有三​​个具体的目标。目标1是识别相关的细胞机制 使用慢性统一的SCI，指导发现新的分子机制以促进 再生。由于 大多数SCI患者（65％）是四肢瘫痪的，手部功能的损失对于 独立性和生活质量。受伤的大鼠在SCI的慢性阶段生存了长达12个月 为了表征神经胶质疤痕，额外的细胞分子（ECM）和轴突状态。目标2是确定 转录机制与慢性构成SCI相关，以指导新发现 分子机制促进再生。我们将使用新开发的技术 在我们的实验室中开发的CRE依赖性核糖量载体，以分离出特异性的mRNA与慢性损伤 CST神经元用于使用RNASEQ表征其转录组的CST神经元。然后我们比较这个 转录组完整且易于损伤的CST神经元（我们已经有这些数据集）和 慢性损伤的CST神经元再生到神经祖细胞（NPC）移植物以识别钥匙 与细胞/轴突生长有关的分子途径被破坏，我们可以治疗靶向 这些分子途径在AIM 3和未来的研究中。目标3是测试候选机制 促进慢性损伤的CST神经元的恢复，并促进轴突再生 慢性科学。我们将首先在AIM2中确定的潜在候选目标进行体外筛选 促进产后皮质神经元和成年背根神经（DRG）神经元的神经突生长。我们 然后测试这些候选基因，以促进长期受伤的CST再生 作为允许性细胞底物的NPC移植。这项工作的发现将大大 扩展我们对慢性SCI的了解，并为慢性SCI开发潜在的治疗方法。

项目成果

Neural Stem Cells: Promoting Axonal Regeneration and Spinal Cord Connectivity.

• DOI: 10.3390/cells10123296
• 发表时间: 2021-11-25
• 期刊: Cells
• 影响因子: 6
• 作者: de Freria CM;Van Niekerk E;Blesch A;Lu P
• 通讯作者: Lu P

Rehabilitation combined with neural progenitor cell grafts enables functional recovery in chronic spinal cord injury.

• DOI: 10.1172/jci.insight.158000
• 发表时间: 2022-08-22
• 期刊: JCI INSIGHT
• 影响因子: 8
• 作者: Lu, Paul;Freria, Camila M.;Graham, Lori;Tran, Amanda N.;Villarta, Ashley;Yassin, Dena;Huie, J. Russell;Ferguson, Adam R.;Tuszynski, Mark H.
• 通讯作者: Tuszynski, Mark H.