Neurobehavioral, cognitive, and mechanistic effects of intranasally administered neural stem cells and environmental enrichment after cortical impact injury in rats

大鼠皮质撞击损伤后鼻内施用神经干细胞和环境富集的神经行为、认知和机制效应

• 批准号: 10468136
• 负责人: MARGARITA GUTOVA
• 金额: $ 50.9万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10468136
• 关键词: 3-Dimensional; Acute; Adult; Advocate; Affect; Affective; Allogenic; Animals; Area; Attention; Behavior; Behavioral; Biological Assay; Brain; Brain Injuries; Brain-Derived Neurotrophic Factor; Cell Lineage; Cell Therapy; Cells; Chronic; Clinic; Clinical; Clinical Trials; Cognition; Cognitive; Combined Modality Therapy; DLG4 gene; DNA; Day Surgery; Dendritic Cells; Distal; Distant; Dose; Dyes; Educational workshop; Equilibrium; Evaluation; Exhibits; Expression Profiling; FOXG1B gene; Female; Glial Fibrillary Acidic Protein; Human; Immune; Immunohistochemistry; Impaired cognition; Individual; Inflammation; Injury; L-myc Gene; Learning; Mediating; Menstrual cycle; Modeling; Morphology; Motor; Natural regeneration; Neurons; Optics; Patients; Pharmacology; Pharmacotherapy; Phase; Pilot Projects; Pre-Clinical Model; Probability; Proteins; Protocols documentation; RNA; Rattus; Recovery; Regenerative capacity; Rehabilitation therapy; Reporting; Signal Transduction; Signaling Molecule; Site; Synapses; TBI Patients; Techniques; Testing; Therapeutic; Time; Tissues; Translating; Translations; Trauma; Traumatic Brain Injury; Traumatic Brain Injury recovery; Treatment Efficacy; Tubulin; Tumorigenicity; United States National Institutes of Health; Walking; analytical method; behavioral outcome; bench to bedside; brain tissue; cell motility; chemokine; clinical translation; clinically relevant; cognitive testing; cytokine; disability; environmental enrichment for laboratory animals; executive function; improved; improved functioning; male; migration; nano-string; nerve stem cell; nestin protein; neural network; neurobehavioral; neurological rehabilitation; neuron loss; neurotrophic factor; neutrophil; preclinical study; regeneration potential; rehabilitation strategy; relating to nervous system; repaired; sex; stem cell therapy; success; water maze; white matter

Traumatic brain injury (TBI) affects more than 10 million individuals worldwide each year and results in long- term motor, cognitive, and affective deficits. Pharmacologic strategies are often used to treat TBI but to date no therapy has successfully translated to the clinic, which advocates for other rehabilitative strategies to restore neuronal networks and recover behavioral deficits thereby increasing the probability of bench-to-bedside success. Neural stem cell (NSC) therapies may be a feasible alternative to pharmacotherapies for improving function after TBI. NSC-based therapies can exploit their inherent ability to migrate to stimulate regeneration and repair damaged brain tissue. In our pilot studies, well-characterized allogeneic human NSCs, LM-NSC008, genetically modified to express the human L-Myc gene were intranasally (IN) administered to adult male and female rats after cortical impact injury. LM-NSC008 cells migrated toward and distributed throughout damaged brain tissue and into distant regions mediating behavioral changes. LM-NSC008 cells significantly improved two distinct cognitive domains - spatial learning (reference learning) and executive function vs. vehicle (VEH). Because clinical translation has been unsuccessful with single therapies, the NIH’s TBI and combination therapy workshop recommended the evaluation of combination treatments. We have reported synergistic benefits when environmental enrichment (EE) is combined with pharmacotherapies and predict augmented benefits with LM-NSC008 cells as well. Our hypotheses are that IN LM-NSC008 cells in male and female rats will 1) migrate and accumulate in sufficient quantities at proximal and distal TBI sites and contribute to behavioral recovery, 2) provide benefit with a clinically relevant delayed administration approach, and 3) improve recovery more robustly when combined with EE than when administered alone. To test our hypotheses, optimize IN delivery doses of LM-NSC008 cells, and to determine LM-NSC008 cell fate and mechanisms, alone and in combination with EE, the following Aims are proposed. Aim 1a: Determine the optimal dose and delivery protocol of IN LM-NSC008 cells for maximal distribution to areas of damage at early, delayed, and chronic time points after TBI. A single high dose of LM-NSC008 cells [6x106] or VEH will be given IN on day-7 (acute period), day-21 (delayed), or day-90 (chronic) after moderate TBI or sham injury, while six lower doses [1x106] will be given once on post-surgery days 7,9,11,13,15,17 (acute), 21,23,25,27,29,31 (delayed), or 90,92,94,96,98,100 (chronic) to determine the protocol that provides maximal distribution of cells at the trauma sites at 3 timepoints after TBI and significantly improves recovery. Aim 1b: Evaluate motor, cognitive, and affective behavioral improvements with IN LM-NSC008 cell therapy in TBI and sham rats. Aim 2: Determine the effect of combining IN LM-NSC008 cell therapy with EE on motor, cognitive, and affective behavior. Aim 3: Determine the fate, mechanisms, and regenerative capacity of IN administered LM-NSC008 cells alone or with EE after TBI.

每年在全球范围内，创伤性脑损伤（TBI）会影响超过1000万个人，并导致长期 术语运动，认知和情感缺陷。药理学策略通常用于治疗TBI，但迄今为止尚未 治疗已成功地转化为诊所，该诊所提倡其他康复策略恢复 神经元网络并恢复行为定义，从而增加了基准对床的概率 成功。神经干细胞（NSC）疗法可能是改善药物疗法的可行替代品 TBI后的功能。基于NSC的疗法可以利用其继承的迁移能力以刺激再生 并修复受损的脑组织。在我们的试点研究中，良好的同种异体人类NSC，LM-NSC008， 通常修改以表达人L-MYC基因的鼻内（IN）给予成年男性和 皮层撞击损伤后的雌性大鼠。 LM-NSC008细胞向迁移并分布在损坏的过程中 脑组织和遥远的区域介导行为变化。 LM-NSC008细胞显着改善 两个不同的认知领域 - 空间学习（参考学习）和执行功能与车辆（车辆）。 由于临床翻译对单疗法不成功，因此NIH的TBI和组合 治疗研讨会建议评估组合治疗。我们报道了协同作用 当环境富集（EE）与药物治疗结合并预测增强时的好处 LM-NSC008细胞也有益。我们的假设是，在LM-NSC008细胞中，男性和雌性大鼠 1）在近端和远端TBI位点迁移并积累了足够数量的 行为恢复，2）通过临床相关的延迟管理方法提供福利，3） 与EE合并时，比单独给药时更强大地改善恢复。测试我们的 假设，优化LM-NSC008细胞的递送剂量，并确定LM-NSC008细胞命运和 单独并与EE结合使用机制，提出了以下目的。目标1a：确定 在LM-NSC008细胞中的最佳剂量和输送方案，以最大程度地分配到早期损害区域 延迟，TBI之后的慢性时间点。将给出一个高剂量的LM-NSC008细胞[6x106]或VEH 在第7天（急性期），第21天（延迟）或中度TBI或假损伤后的第90天（慢性），而六个 较低的剂量[1x106]将在手术后一次给予一次7,9,11,13,15,17（急性），21,23,25,27,29,31 （延迟），或90,92,94,96,98,100（慢性），以确定提供细胞最大分布的方案 在TBI后3个时间点的创伤部位，在创伤部位并显着改善了恢复。 AIM 1B：评估电动机， TBI和假大鼠的LM-NSC008细胞疗法中的认知和情感行为改善。目标2： 确定在LM-NSC008细胞疗法中与EE组合对运动，认知和情感的影响 行为。目标3：确定管理LM-NSC008中的命运，机制和再生能力 单独或在TBI之后使用EE。