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.

在军事人员中，创伤性脑损伤（TBI）的发生率不成比例。甚至温和 TBI可以具有持久的不良神经生物学作用，最初受伤后可以继续发展。 癫痫发作，痉挛，认知和情感障碍，睡眠问题和内分泌疾病（糖尿病）是 普遍，令人衰弱，难以治疗。有效的长期康复将需要变革 治疗方法解决了导致延迟症状出现的大脑过程。 TBI后这些问题的延迟演变提供了预防和改善长期的机会 结果。我们开发的基因输送技术产生了基因的持续过表达 离散大脑区域中的神经肽生长抑素（SST）。使用腺相关的颅内基因递送 与我们的SST载体相似的病毒（AAV）向量在几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，多种机制可能会提供阳性 延迟TBI效应的好处。我们的SST基因递送到海马可预防电气 在已建立的癫痫模型中测试的70％的大鼠中发育的诱发癫痫发作。下一步 将这种方法转换为临床上有用的应用是在更先进的大脑动物模型中对其进行测试 受伤。同时，确定此方法的安全性对于进一步的临床前至关重要 发展。因此，功效和安全性包括这个小型项目的2个特定目的，旨在用作 转化进度基金会。测试功效是否扩展到癫痫发作的延迟发展 在TBI之后，我们将结合闭合头TBI的特征良好的大鼠模型与点燃模型 首先观察到哪些功效。麻醉的年轻雄性大鼠将受控脑损伤10 在接受海马中的SST或对照基因转移载体的颅内输注之前的几天， 在单手术期间，永久植入电极进行电刺激。一周后 使用强度子阈值，每天将两次定时刺激启动程序 癫痫发作。配对的脑电图（EEG）和刺激期间获得的视频记录 随着严重程度和持续时间的重复措施，癫痫发作将盲目离线评分。逐渐在 随后几天到几周，轻度癫痫发作开始并变得更加严重，直到完全达到 点燃的状态，刺激始终引起未处理大鼠的最大强度癫痫发作。 海马SST基因递送的治疗功效将反映在癫痫发作严重程度降低或 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 一贯唤起。检查TBI，基因转移和点燃对内存性能敏感的影响 为了海马受伤，大鼠探索Y形迷宫交替臂的自然趋势是连续的 在整个研究过程中，试验将反复测试。认知表现将在多个 挑战对学习和记忆能力敏感的任务。天然运动功能（活动，饲养，修饰）， 情感状态（焦虑）和昼夜节律生理学（睡眠）将从连续的家庭笼子进行评估 红外视频。治疗安全性将从全面的行为评估中进行评估，但也将通过 脑病理学随载体治疗的函数的全面组织学分析。此外 病理标记，我们将评估生物tbi大鼠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和进行改善 TBI后认知和运动功能障碍将涉及多种疗效机制 大脑特定子区域中的受体亚型。这个可行且创新的小型项目开放了新的 在TBI之后，改善了面临数十年衰弱后果的退伍军人的康复途径。