Intranasal delivery of neural stem cells alone or in combination with environmental enrichment as a potential therapy for experimental TBI

创伤性脑损伤每年影响全球1000多万人，是导致脑损伤的主要原因。 致残率高，导致脑外伤幸存者长期功能缺陷。到目前为止，治疗的主要未满足需求 颅脑损伤是恢复神经元网络和恢复功能的有效策略。虽然药理上 策略是治疗脑外伤最常见的方法，它们和其他方法一样，受到血-脑--的阻碍。 屏障(BBB)渗透性。因此，需要一种绕过血脑屏障的治疗方法。神经干 基于细胞(NSC)的治疗可能是改善脑外伤后功能的药物治疗的一种可行的替代方案。 然而，基于干细胞的治疗取决于有效地将药物输送到受损区域。在我们的试验性TBI中 研究表明，具有良好特性的同种异体人神经干细胞经基因修饰后表达人L-Myc基因(Lm-Myc)。 NSC008)迁移到受损脑区并分布于受损脑区，大鼠在 接受鼻腔输送的神经干细胞。我们假设1)鼻腔递送的LM-NSC008神经干细胞将 迁移到脑损伤部位，积累足够的量，有助于运动和认知的恢复 损伤后，和2)增加神经干细胞与环境丰富(EE)将提供进一步的好处。 为了检验我们的假设并优化NSC的交付，我们提出了三个目标。目标1：确定最优 神经干细胞给药的剂量，使其最大限度地分布到脑外伤损伤区域。我们将测试两种剂量范例：一种 两个剂量(6x106或12x106神经干细胞)或赋形剂将在中度颅脑损伤或假伤后第7天给予， 或较低剂量(1x106或2x106神经干细胞)将从术后第7天开始隔天给药至第二天。 17.应用于光学透明脑切片的计算分析方法(清晰度技术)将是 用于量化和验证TBI与Sham中的NSC迁移和分布。NSC分布的特征 在3D组织中结合路径查找算法将使我们能够预测NSC剂量依赖的路径 迁移和脑组织的生物分布。目的2：确定神经干细胞疗法改善运动能力的程度 结果和认知(参考记忆和执行功能)。神经干细胞的最佳剂量，由 在损伤部位的最大分布，从目标1将被给予不同的队列的大鼠在其中之一 伤后3次(7d[急性期]、21d[延迟期]和3mo[慢性期])。目标 3：确定EE联合NSC治疗对运动和认知功能的影响。NSC方案 在Aim 2中使用的将与临床相关的每天4小时EE的康复范例相结合，我们 已经进行了优化，以模拟患者在诊所的时间。EE接收组将与标准住房进行比较 目标2中的(非强化)组。我们预计这一组合将在改善运动和 认知功能优于NSC单独治疗。所有实验将包括雄性和正常骑自行车的雌性大鼠。 实现特定目标有可能极大地改善脑外伤的治疗。 并可能影响其他神经退行性疾病的治疗模式。

Traumatic brain injury (TBI) affects more than 10 million individuals worldwide each year and is a major cause of disability, resulting in long-term functional deficits for TBI survivors. To date, the major unmet needs for treating TBI are effective strategies to restore neuronal networks and recover function. Although pharmacologic strategies are the most common approach to treat TBI, they, and other approaches, are hindered by blood-brain- barrier (BBB) permeability. Therefore, a therapeutic approach that circumvents the BBB is needed. Neural stem cell (NSC)-based therapies may be a feasible alternative to pharmacotherapies for improving function after TBI. However, stem cell-based therapies are contingent on efficient delivery to the areas of damage. In our pilot TBI studies, well-characterized allogeneic human NSCs genetically modified to express the human L-Myc gene (LM- NSC008) migrated to and distributed at damaged brain regions, and rats showed improved spatial learning after receiving intranasally delivered NSCs. We hypothesize that 1) intranasally-delivered LM-NSC008 NSCs will migrate to TBI sites, accumulate in sufficient quantities, and contribute to motor and cognitive recovery post-injury, and 2) augmenting NSCs with environmental enrichment (EE) will provide further benefits. To test our hypothesis and optimize NSC delivery, we propose three Aims. Aim 1: To determine the optimal dose of NSC delivery for maximal distribution to areas of TBI damage. We will test two dosing paradigms: one of two bolus doses (6x106 or 12x106 NSCs) or vehicle will be given on day 7 after moderate TBI or sham injury, or a lower dose (1x106 or 2x106 NSCs) will be given on alternate days starting on day 7 post-surgery through day 17. Computational analytical methods applied to optically-cleared brain sections (CLARITY technique) will be used to quantify and validate NSC migration and distribution in TBI vs. shams. Characterizing NSC distribution in 3D tissue combined with route finding algorithms will allow us to predict NSC dosing-dependent routes of migration and brain tissue biodistribution. Aim 2: To determine the extent to which NSC therapy improves motor outcome and cognition (reference memory and executive function). The optimal dose of NSCs, determined by the greatest distribution at the site of injury, from Aim 1 will be administered to separate cohorts of rats at one of three times (7-d [acute phase], 21-d [delayed phase], and 3-mo [chronic phase]) after TBI or sham injury. Aim 3: To determine the effect of combining EE with NSC therapy on motor and cognitive benefits. The NSC regimen used in Aim 2 will be combined with a clinically-relevant rehabilitation paradigm of 4 h of EE per day, which we have optimized to mimic patient time in the clinic. The EE receiving groups will be compared to standard-housed (non-enriched) groups in Aim 2. We expect that the combination will be more effective at improving motor and cognitive function than NSC therapy alone. All experiments will include male and normal cycling female rats. Achieving the specific aims has the potential to make tremendously improve the treatment of TBI patients and potentially impact therapeutic paradigms for other neurodegenerative diseases.