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

创伤性脑损伤每年影响全球1000多万人，并导致长期的脑损伤。 运动、认知和情感缺陷一词。治疗颅脑损伤常采用药物治疗策略，但迄今未见 治疗已经成功地转化到诊所，该诊所倡导采取其他康复策略来恢复 神经元网络和恢复行为缺陷，从而增加卧床的可能性 成功。神经干细胞(NSC)疗法可能是药物疗法的一种可行的替代方案，以改善 TBI后的功能。基于神经干细胞的疗法可以利用其固有的迁移能力来刺激再生 修复受损的脑组织。在我们的初步研究中，具有良好特性的同种异体人类神经干细胞，LM-NSC008， 将表达人L-MYC基因的转基因动物经鼻腔给药给成年男性和 雌性大鼠皮质撞击伤后。Lm-NSC008细胞向损伤方向迁移并分布于损伤部位 脑组织和远距离区域，调节行为的变化。Lm-NSC008细胞显著改善 两个截然不同的认知域--空间学习(参照学习)和执行功能与载体(VEH)。 由于单一疗法的临床翻译不成功，美国国立卫生研究院的脑损伤和联合疗法 治疗工作坊推荐联合治疗方案的评价。我们已经报告了协同效应 环境浓缩(EE)与药物治疗相结合的益处和预测增强 使用LM-NSC008细胞也有好处。我们的假设是在雄性和雌性大鼠的Lm-NSC008细胞中 1)是否会在TBI近端和远端有足够数量的迁移和积累，并有助于 行为恢复，2)提供临床上相关的延迟给药方法，以及3) 与单独使用EE相比，联合使用EE可更有力地改善康复。测试我们的 假设，优化LM-NSC008细胞的递送剂量，并确定LM-NSC008细胞的命运和 机制，单独和与环境工程结合，提出了以下目标。目标1a：确定 IN LM-NSC008细胞在早期最大限度地分布到损伤区域的最佳剂量和给药方案， 颅脑损伤后延迟、慢性时间点。将给予单次高剂量的LM-NSC008细胞[6x106]或VEH 在中度颅脑损伤或假伤后的第7天(急性期)、第21天(延迟)或第90天(慢性)，而6天 术后第7天、第9天、第11天、第13天、第15天、第17天(急性)、第21、23、25、27、29、31天将给予较低剂量[1x106]一次。 或90，92，94，96，98,100(慢性)，以确定提供最大细胞分布的方案 在伤后3个时间点，均可明显改善伤情恢复。目标1b：评估电机， IN LM-NSC008细胞疗法改善了脑外伤和假手术大鼠的认知和情感行为。目标2： 确定在LM-NSC008细胞治疗中联合EE对运动、认知和情感的影响 行为。目的3：确定IN给药的LM-NSC008的命运、机制和再生能力 脑损伤后单独或伴有EE的细胞。

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.