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: 9026505
• 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 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9026505
• 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.

创伤性脑损伤（TBI）的发病率在军事人员中不成比例地高。即使是轻微 TBI可能具有持续的不良神经生物学影响，这些影响可能在初始损伤后很长时间内继续发展。 癫痫、痉挛、认知和情感障碍、睡眠问题和内分泌疾病（糖尿病）是 流行，衰弱，难以治疗。有效的长期康复将需要变革 治疗方法，解决负责出现延迟症状的大脑过程。 TBI后这些问题的延迟发展为预防和改善长期预后提供了机会。 结果。我们开发的一种基因传递技术可以使基因持续过度表达， 神经肽生长抑素（SST）在离散的大脑区域。使用腺相关病毒介导的颅内基因递送 与我们的SST载体类似的病毒（AAV）载体在几种脑组织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， 情感功能，镇痛，细胞保护，多种机制，它可能提供积极的 对延迟TBI效应的好处。我们将SST基因传递到海马体， 在已建立的癫痫模型中，70%的大鼠出现癫痫发作。的下一步 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 损伤同时，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，有效性和安全性构成了本小型项目的2个具体目标， 转化进步的基础。为了测试有效性是否延伸到癫痫发作的延迟发展 在TBI后，我们将联合收割机将闭合头TBI的良好表征的大鼠模型与点燃癫痫模型结合， 首次观察到该功效。将麻醉的年轻成年雄性大鼠给予受控的脑损伤10 在海马中接受SST或对照基因转移载体的颅内输注前30天，和 用于电刺激的永久性植入电极，在单次手术期间。一周后， 将启动程序，包括每天两次定时刺激，使用强度低于阈值， 癫痫发作一代。刺激期间获得的成对脑电图（EEG）和视频记录 将对癫痫发作进行盲态离线评分，作为严重程度和持续时间的重复测量。逐年 几天到几周后，轻度癫痫发作开始，并逐渐变得更加严重，直到达到完全发作。 点燃状态，其中刺激一致地激发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，癫痫严重程度进展延迟，或可能的最大癫痫严重程度降低 不断地唤起。研究TBI、基因转移和点燃对记忆表现敏感性的影响， 海马损伤，大鼠探索Y形迷宫交替臂的自然倾向， 在整个研究过程中将反复进行试验。认知表现将在多个 挑战任务对学习和记忆能力敏感。自然运动功能（活动，饲养，梳理）， 将在连续饲养笼中评估情感状态（焦虑）和昼夜生理学（睡眠） 红外视频治疗安全性将通过综合行为评估进行评价，但也通过 作为载体治疗的功能的脑病理学的综合组织学分析。除了 病理学标记，我们将评估基因转移在点燃TBI大鼠的空间定位的影响 SST受体蛋白的模式。我们建议，减少癫痫发作，改善进行性 TBI后认知和运动功能障碍，将涉及多种功效机制， 大脑特定亚区的受体亚型。这一创新性的小项目，开启了新的 改善退伍军人在TBI后面临数十年衰弱后果的康复途径。