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: 10688136
• 负责人: MARGARITA GUTOVA
• 金额: $ 51.03万
• 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/10688136
• 关键词: 3-Dimensional; Acceleration; 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; 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; Intranasal Administration; L-myc Gene; Learning; Mediating; Menstrual cycle; Modeling; Morphology; Motor; Natural regeneration; Neurons; Optics; Patients; Pharmacotherapy; Phase; Pilot Projects; Pre-Clinical Model; Probability; Proteins; Protocols documentation; RNA; Rattus; Recommendation; Recovery; Regenerative capacity; Rehabilitation therapy; Reporting; Signal Transduction; Signaling Molecule; Site; Synapses; Synapsins; 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; Visualization; Walking; analytical method; behavioral outcome; bench to bedside; brain tissue; cell motility; chemokine; clinical translation; clinically relevant; cognitive testing; cytokine; disability; efficacy evaluation; environmental enrichment for laboratory animals; executive function; functional improvement; glial activation; improved; male; migration; nano-string; nerve stem cell; nestin protein; neural; neural network; neurobehavioral; neurological rehabilitation; neuron loss; neurotrophic factor; neutrophil; pharmacologic; preclinical study; regeneration potential; 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.

创伤性脑损伤 (TBI) 每年影响全球超过 1000 万人，并导致长期 术语运动、认知和情感缺陷。药物策略通常用于治疗 TBI，但迄今为止还没有 疗法已成功转化为临床，提倡采用其他康复策略来恢复 神经元网络并恢复行为缺陷，从而增加从实验室到临床的可能性 成功。神经干细胞（NSC）疗法可能是药物疗法的可行替代方案，以改善 TBI 后的功能。基于 NSC 的疗法可以利用其固有的迁移能力来刺激再生 并修复受损的脑组织。在我们的试点研究中，充分表征的同种异体人类 NSC，LM-NSC008， 经基因改造以表达人类 L-Myc 基因的药物被鼻内 (IN) 施用给成年男性和 皮质撞击损伤后的雌性大鼠。 LM-NSC008 细胞迁移并分布在受损区域 脑组织和远处区域介导行为变化。 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（VEH）。 由于单一疗法的临床转化并不成功，NIH 的 TBI 和联合疗法 治疗研讨会建议对联合治疗进行评估。我们报告了协同作用 当环境丰富（EE）与药物疗法相结合时的好处并预测增强 LM-NSC008 细胞也有好处。我们的假设是雄性和雌性大鼠的 IN LM-NSC008 细胞 将 1) 在近端和远端 TBI 部位迁移并积累足够数量，并有助于 行为恢复，2) 通过临床相关的延迟给药方法提供益处，以及 3) 与单独使用 EE 联合使用时，可以更有效地改善恢复。来测试我们的 假设，优化 LM-NSC008 细胞的 IN 递送剂量，并确定 LM-NSC008 细胞的命运和 单独或与 EE 结合的机制，提出了以下目标。目标 1a：确定 IN LM-NSC008 细胞的最佳剂量和递送方案，可在早期最大程度地分布到损伤区域， TBI 后的延迟和慢性时间点。将给予单次高剂量 LM-NSC008 细胞 [6x106] 或 VEH 中度 TBI 或假伤后第 7 天（急性期）、第 21 天（延迟期）或第 90 天（慢性期）IN，而 6 较低剂量 [1x106] 将在术后第 7、9、11、13、15、17（急性）、21、23、25、27、29、31 天给予一次 （延迟）或 90,92,94,96,98,100（慢性）以确定提供最大细胞分布的方案 TBI 后 3 个时间点在创伤部位进行治疗，并显着改善恢复。目标 1b：评估电机， IN LM-NSC008 细胞疗法可改善 TBI 和假大鼠的认知和情感行为。目标 2： 确定 IN LM-NSC008 细胞疗法与 EE 相结合对运动、认知和情感的影响 行为。目标 3：确定 IN 施用 LM-NSC008 的命运、机制和再生能力 TBI 后单独使用细胞或与 EE 一起使用细胞。