Somatostatin gene delivery to enhance long-term functional recovery from TBI

生长抑素基因递送可增强 TBI 的长期功能恢复

• 批准号: 8732764
• 负责人: MICHAEL A KING
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8732764
• 关键词: Absence of pain sensation; Address; Adverse effects; Affect; Affective; Amygdaloid structure; Animal Model; Animals; Anxiety; Astrocytosis; Autopsy; Behavior assessment; Behavioral; Biological Assay; Brain; Brain Diseases; Brain Injuries; Brain Pathology; Brain region; Caring; Circadian Rhythms; Clinical; Clinical Trials; Cognition; Cognition Disorders; Cognitive; Complement; Cytoprotection; Development; Diabetes Mellitus; Disease; Electric Stimulation; Electrodes; Emotional; Endocrine System Diseases; Ensure; Epilepsy; Epileptogenesis; Evaluation; Evolution; Experimental Designs; Experimental Models; Foundations; Functional disorder; Gene Delivery; Gene Expression; Gene Transfer; Generations; Goals; Grooming; Head; Health; Health Benefit; Healthcare; Hippocampus (Brain); Home environment; Impaired cognition; Implanted Electrodes; Incidence; Individual; Inflammation; Infusion procedures; Injury; Intractable Epilepsy; Investigation; Kindling (Neurology); Learning; Long-Term Care; Measures; Mediating; Memory; Methods; Military Personnel; Modeling; Mood Disorders; Morbidity - disease rate; Motor; Motor Activity; Neuraxis; Neurobiology; Neurodegenerative Disorders; Neurologic; Neurons; Neuropeptide Gene; Neuropeptides; Operative Surgical Procedures; Outcome; Outcome Measure; Pathology; Patients; Pattern; Performance; Physiological; Physiology; Pre-Clinical Model; Prevention; Procedures; Process; Protocols documentation; Quality of life; Rattus; Recovery; Recovery of Function; Rehabilitation therapy; Research; Rodent Model; Safety; Seizures; Services; Severities; Shapes; Sleep; Sleep Disorders; Somatostatin; Somatostatin Receptor; Surveys; Symptoms; Syndrome; Techniques; Testing; Therapeutic; Time; Translating; Traumatic Brain Injury; Treatment Efficacy; Veterans; Video Recording; Viral; adeno-associated viral vector; analog; anxiety states; arm; behavior test; blind; cognitive performance; cost; disability; efficacy testing; experience; functional outcomes; gene therapy; gene transfer vector; high risk; immunoreactivity; improved; injured; innovation; long-term rehabilitation; male; mild traumatic brain injury; model development; motor disorder; neuron loss; neuropathology; neuropsychiatry; neuropsychological; novel; novel therapeutic intervention; novel therapeutics; pre-clinical; prevent; public health relevance; receptor; receptor expression; response; synaptic inhibition; treatment effect; vector; vector control; viral gene delivery; young adult

The incidence of traumatic brain injury (TBI) is disproportionately high among military personnel. Even mild TBI can have persistent adverse neurobiological effects that can continue to evolve long after the initial injury. Seizures, spasticity, cognitive and affective disorders, sleep problems, and endocrine diseases (diabetes) are prevalent, debilitating, and difficult to treat. Effective long-term rehabilitation will require transformative therapeutic approaches that address the brain processes responsible for the emergence of delayed symptoms. The delayed evolution of these problems post-TBI offers opportunities for prevention and improved long-term outcome. A gene delivery technique we developed produces sustained over-expression of the gene for neuropeptide somatostatin (SST) in discrete brain regions. Intracranial gene delivery using adeno-associated viral (AAV) vectors similar to our SST vector is proving safe and effective in clinical trials for several brain diseases. In the central nervous system SST participates in synaptic inhibition, inflammation, cognition, emotional function, analgesia, and cytoprotection, multiple mechanisms by which it might provide positive benefits against delayed TBI effects. Our SST gene delivery to the hippocampus prevented electrically induced seizures from developing in 70% of rats tested in an established epilepsy model. The next step in translating this approach to clinically useful applications is to test it in more advanced animal models of brain injury. At the same time, determining the safety of this method will be essential for further preclinical development. Efficacy and safety thus comprise 2 specific aims of this Small Project intended to serve as a foundation for translational progress. To test whether efficacy extends to the delayed development of seizures after TBI, we will combine a well-characterized rat model for closed-head TBI with the kindling seizure model in which efficacy was first observed. Anesthetized young adult male rats will be given a controlled brain injury 10 days before receiving intracranial infusion of SST or control gene transfer vectors in hippocampus, and permanently implanted electrodes for electrical stimulation, during a single surgery. A week later a kindling procedure will be initiated composed of timed stimulation twice per day, using intensities sub-threshold for seizure generation. Paired electroencephalograph (EEG) and video recordings obtained during stimulated seizures will be scored blind offline as duplicate measures of severity and duration. Gradually over the following days to weeks mild seizures start and become progressively more severe until reaching a fully kindled state where the stimulation consistently elicits maximally intense seizures in untreated rats. Therapeutic efficacy of hippocampal SST gene delivery will be reflected in reduced seizure severity or duration, delayed progression of seizure severity, or a reduction in the maximal seizure severity that can be consistently evoked. To examine effects of TBI, gene transfer, and kindling on memory performance sensitive to hippocampal injury, a natural tendency of rats to explore alternating arms of a Y-shaped maze on successive trials will be tested repeatedly throughout the study. Cognitive performance will be evaluated on multiple challenge tasks sensitive to learning and memory ability. Natural motor function (activity, rearing, grooming), affective states (anxiety), and circadian physiology (sleep) will be assessed from continuous home cage infrared video. Therapeutic safety will be evaluated from comprehensive behavioral assessments, but also by comprehensive histological analysis of brain pathology as a function of vector treatment. In addition to markers for pathology, we will evaluate the effects of gene transfer in kindled TBI rats on spatial localization patterns of SST receptor proteins. We propose that seizure reduction, and amelioration of progressive cognitive and motor dysfunction post-TBI, will involve multiple efficacy mechanisms that engage several receptor subtypes in specific subregions of the brain. This feasible and innovative Small Project opens new avenues for improving the rehabilitation of Veterans facing decades of debilitating consequences after TBI.

军事人员中创伤性脑损伤（TBI）的发生率异常高。甚至温和 TBI 可能会产生持续的不良神经生物学影响，这种影响在初次损伤后很长时间内仍会继续发展。 癫痫、痉挛、认知和情感障碍、睡眠问题和内分泌疾病（糖尿病） 普遍存在、使人衰弱且难以治疗。有效的长期康复需要变革 针对导致延迟症状出现的大脑过程的治疗方法。 TBI 后这些问题的延迟发展为预防和长期改善提供了机会 结果。我们开发的基因传递技术可产生持续的基因过度表达 离散大脑区域中的神经肽生长抑素（SST）。使用腺相关基因进行颅内基因传递 与我们的 SST 载体相似的病毒 (AAV) 载体在多个大脑的临床试验中被证明是安全有效的 疾病。在中枢神经系统中，SST 参与突触抑制、炎症、认知、 情绪功能、镇痛和细胞保护，它可能提供积极的多种机制 对抗延迟 TBI 效应的益处。我们的 SST 基因传递到海马体阻止了电 在已建立的癫痫模型中测试的大鼠中有 70% 诱发癫痫发作。下一步在 将这种方法转化为临床有用的应用是在更先进的大脑动物模型中进行测试 受伤。同时，确定该方法的安全性对于进一步的临床前研究至关重要 发展。因此，功效和安全性构成了这个小型项目的 2 个具体目标，旨在作为 转化进步的基础。测试疗效是否延伸至癫痫发作延迟发展 TBI 后，我们将结合良好表征的闭头 TBI 大鼠模型与点燃癫痫发作模型 首次观察到其功效。麻醉的年轻成年雄性大鼠将受到控制性脑损伤 10 在海马区接受 SST 或对照基因转移载体颅内输注前几天，以及 在单次手术中永久植入用于电刺激的电极。一周后点燃 程序将开始，包括每天两次定时刺激，使用强度亚阈值 癫痫发作的一代。配对脑电图 (EEG) 和刺激过程中获得的视频记录 癫痫发作将离线进行盲法评分，作为严重程度和持续时间的重复测量。逐渐过了 接下来的几天到几周，轻微的癫痫发作开始并逐渐变得更加严重，直到完全发作。 点燃状态，其中刺激持续引起未经治疗的大鼠最强烈的癫痫发作。 海马 SST 基因递送的治疗效果将反映在癫痫发作严重程度的减轻或 持续时间、癫痫发作严重程度的延迟进展或最大癫痫发作严重程度的降低 持续地被唤起。检查 TBI、基因转移和点燃对记忆表现敏感的影响 海马损伤是大鼠连续探索 Y 形迷宫交替臂的自然倾向 在整个研究过程中将反复测试试验。认知表现将通过多种方式进行评估 挑战对学习和记忆能力敏感的任务。自然运动功能（活动、饲养、梳理）， 情感状态（焦虑）和昼夜节律生理学（睡眠）将在连续的笼子中进行评估 红外视频。治疗安全性将通过综合行为评估进行评估，但也通过 脑病理学的综合组织学分析作为媒介治疗的功能。此外 病理学标记，我们将评估点燃 TBI 大鼠的基因转移对空间定位的影响 SST受体蛋白的模式。我们建议减少癫痫发作并改善进行性 TBI 后的认知和运动功能障碍将涉及多种功效机制，涉及多种因素 大脑特定分区中的受体亚型。这个可行且创新的小项目开启了新的 改善退伍军人康复的途径，这些退伍军人在创伤性脑损伤后面临数十年的衰弱后果。