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（TBI）每年影响全球超过1000万人，并导致长期的脑损伤。 长期运动、认知和情感缺陷。药物策略通常用于治疗TBI，但迄今为止， 治疗已成功地转化为诊所，倡导其他康复策略，以恢复 神经元网络和恢复行为缺陷，从而增加的可能性， 成功神经干细胞（NSC）疗法可能是一种可行的替代药物治疗，以改善 TBI后的功能基于神经干细胞的治疗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，充分表征的同种异体人NSC，LM-NSC 008， 将经遗传修饰以表达人L-Myc基因的人L-Myc基因鼻内（IN）给予成年男性， 皮质撞击损伤后的雌性大鼠。LM-NSC 008细胞向受损细胞迁移并分布在受损细胞中， 脑组织和远距离区域介导的行为变化。LM-NSC 008细胞显著改善 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不成功，NIH的TBI和联合疗法 治疗研讨会建议评估联合治疗。我们已经报道了协同作用 当环境富集（EE）与药物治疗相结合时， LM-NSC 008细胞也有好处。我们的假设是，在雄性和雌性大鼠中， 将1）在近端和远端TBI部位迁移和累积足够的量，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EE联合给药时比单独给药时更有力地改善恢复。来测试我们 假设，优化LM-NSC 008细胞的IN递送剂量，并确定LM-NSC 008细胞命运， 机制，单独和与EE相结合，提出了以下目标。目标1a：确定 IN LM-NSC 008细胞的最佳剂量和递送方案， TBI后延迟和慢性时间点。将给予单次高剂量LM-NSC 008细胞[6x 106]或VEH 在中度TBI或假损伤后第-7天（急性期）、第-21天（延迟期）或第-90天（慢性期）IN， 将在术后第7、9、11、13、15、17（急性）、21、23、25、27、29、31天给予一次较低剂量[1x 106] （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 在创伤部位在TBI后3个时间点，并显着改善恢复。目标1b：评估电机， 在TBI和假手术大鼠中用IN LM-NSC 008细胞疗法改善认知和情感行为。目标二： 确定IN LM-NSC 008细胞疗法与EE联合对运动、认知和情感的影响 行为目的3：确定IN施用的LM-NSC 008的命运、机制和再生能力 细胞单独或与TBI后EE。

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.