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

单独鼻内递送神经干细胞或与环境富集相结合作为实验性 TBI 的潜在疗法

• 批准号: 10265107
• 负责人: MARGARITA GUTOVA
• 金额: $ 42.99万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10265107
• 关键词: 3-Dimensional; Acute; Affect; Algorithms; Allogenic; Area; Attention; Behavioral; Biodistribution; Blood - brain barrier anatomy; Bolus Infusion; Brain; Brain Injuries; Brain region; Cell Therapy; Cells; Chronic Phase; Clinic; Clinical; Cognition; Cognitive; Cognitive deficits; Data; Dose; Estrus; Female; Functional disorder; Growth Factor; Hormones; Human; Individual; Injury; Investigational Therapies; L-myc Gene; Learning; Memory; Modeling; Motor; Natural regeneration; Nervous system structure; Neurodegenerative Disorders; Neuroglia; Neurons; Operative Surgical Procedures; Optics; Outcome; Paper; Patients; Pharmacology; Pharmacotherapy; Phase; Pilot Projects; Population; Protocols documentation; Publishing; Rattus; Recovery; Regimen; Rehabilitation therapy; Replacement Therapy; Route; Schedule; Site; Survivors; TBI Patients; TBI treatment; Techniques; Testing; Therapeutic; Therapeutic Agents; Time; Tissues; Translations; Trauma; Traumatic Brain Injury; Traumatic Brain Injury recovery; Treatment Efficacy; analytical method; blood-brain barrier permeabilization; brain tissue; cell injury; cell motility; cell type; clinical translation; clinically relevant; cognitive benefits; cognitive function; cognitive recovery; cohort; controlled cortical impact; cost effective; disability; environmental enrichment for laboratory animals; executive function; experimental study; genetically modified cells; immunoregulation; improved; improved functioning; injury recovery; male; migration; motor deficit; motor function improvement; motor recovery; nerve stem cell; nervous system disorder; neurogenesis; neurological rehabilitation; neuroprotection; neurotrophic factor; rehabilitation paradigm; repaired; sex; small molecule; stem cell therapy; stem cells; symptom treatment

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.

创伤性脑损伤（TBI）每年影响全球1000多万人，是导致脑损伤的主要原因。 残疾，导致TBI幸存者的长期功能缺陷。迄今为止，治疗的主要未满足需求 TBI是恢复神经网络和功能的有效策略。虽然药理学 策略是治疗TBI的最常见方法，它们和其他方法都受到血脑屏障的阻碍， 屏障（BBB）通透性。因此，需要一种绕过BBB的治疗方法。神经干 细胞（NSC）为基础的治疗可能是一个可行的替代药物治疗改善功能后TBI。 然而，基于干细胞的疗法取决于有效地递送到损伤区域。在我们的试点TBI中， 研究中，充分表征的同种异体人类神经干细胞基因修饰表达人类L-Myc基因（LM-1）， NSC 008）迁移到并分布在受损的脑区，并且大鼠表现出改善的空间学习， 接受鼻内递送的NSC。我们假设1）鼻内递送的LM-NSC 008 NSC将 迁移到TBI部位，积累足够的量，并有助于运动和认知恢复 2）用环境富集（EE）增强NSC将提供进一步的益处。 为了检验我们的假设和优化NSC交付，我们提出了三个目标。目标1：确定最佳 NSC输送剂量，以最大限度地分布到TBI损伤区域。我们将测试两种给药模式： 在中度TBI或假损伤后第7天给予两次推注剂量（6 × 106或12 × 106个NSC）或载体， 或从术后第7天开始隔日给予较低剂量（1 × 106或2 × 106个NSC），直至第10天 17.将应用于光学透明脑切片的计算分析方法（显微镜技术） 用于量化和验证TBI与假手术中的NSC迁移和分布。描述NSC分布 在3D组织中结合路由查找算法将使我们能够预测NSC剂量依赖性路由， 迁移和脑组织生物分布。目的2：确定NSC治疗在多大程度上改善运动功能 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开始，将在以下之一给予单独的大鼠队列： TBI或假损伤后3次（7天[急性期]、21天[延迟期]和3个月[慢性期]）。目的 3：确定EE与NSC治疗相结合对运动和认知益处的影响。NSC方案 目标2中使用的药物将与每天4小时EE的临床相关康复模式相结合， 优化了模拟病人在诊所的时间。EE接收组将与标准圈养组进行比较。 目标2中的（非富集）组。我们预计，这种组合将更有效地改善电机和 认知功能优于单独的NSC治疗。所有实验将包括雄性和正常骑自行车的雌性大鼠。 这些目标的实现有可能极大地改善TBI的治疗 患者，并可能影响其他神经退行性疾病的治疗模式。