Osmotic therapy for seizures in pediatric traumatic brain injury

 描述(由申请者提供)：这个有指导的临床科学家研究职业发展奖(K08)的目标是：a)获得以下方面的技能：活体多光子成像，处理和培养器官型脑片的经验，以及记录啮齿动物的脑电。B)拥有出版物和初步数据，以便在获奖的第4年前提交强有力的R01申请。我的最终目标是成为抑制生理学和儿童癫痫的独立研究员，特别是与创伤性脑损伤(TBI)相关的研究。在儿童年龄组中，脑外伤是导致发病率和死亡率的主要原因，导致超过50%的儿童死亡。癫痫发作是儿童颅脑损伤的常见并发症。脑外伤是如何导致癫痫发作的尚不清楚。几种病理条件，包括脑损伤，导致神经元氯离子浓度升高，[Cl-]i在神经元中非常重要，因为当[Cl-]i低于神经元的静息膜电位时，通过GABAA受体的Cl-流量介导抑制，但当[Cl-]i高于静息膜电位时，[Cl-]i可导致兴奋。我们最新的研究表明，在神经科重症监护病房用于治疗脑水肿的甘露醇和渗透剂，不仅减少了神经元的体积，而且还降低了[Cl-]i。因此，甘露醇应该具有抗惊厥的特性。我的建议旨在回答以下假设：甘露醇(一种典型的高渗药物)由于[Cl-]i的减少而减少了新皮质的癫痫样活动，这种[Cl-]i的减少导致了GABAAR介导的抑制效应的增加。为了回答这一假设，我们将使用体外/体内多光子成像和电生理技术在表达一种新的氯敏感放射性荧光团(SuperClomeleon)的转基因小鼠中进行。通过课程和实践经验，K08应用程序的目标是获得以下方面的必要技能：A)多光子活体成像；B)处理和培养器官型脑片；C)记录和分析啮齿动物的脑电。这些技术将使我能够完成我提议的项目，并给我所需的技术知识，以补充我目前的体外电生理学技能。我在基础研究和临床儿科神经学方面有很强的背景，这让我能够理解和看到家庭由于创伤后癫痫治疗的局限性而经历的绝望。此外，我发表的11篇原创论文以及每家资助机构至少交付一篇论文的竞争性资助记录证明了我的高度动机、完成项目的能力以及继续从事神经抑制生理学和癫痫研究的愿望。我是麻省总医院神经内科儿科神经科的全职教员，也是哈佛医学院神经病学的讲师。我得到了神经科主任和儿科神经科主任的支持，成为MGH的一名富有成效的独立研究员。凭借MGH的尖端技术和设施(双光子显微镜、电生理设备)，拥有出色记录的导师(Staley博士、Bacskai博士)，一个讨论我的研究进展和翻译可能性的咨询委员会(Rosand博士、Duhaime博士、Cash博士)，MGH的协作氛围，以及作为哈佛医学院的一部分，我处于一个极佳的环境中，可以作为一名独立研究员发展一份强大、多产和有价值的职业生涯。K08培训将是帮助我作为一名独立调查员取得成功的基础。

 DESCRIPTION (provided by applicant): The goal of this Mentored Clinical Scientist Research Career Developmental Award (K08) is to: A) Obtain skills in: in vivo multiphoton imaging, experience working with and culturing organotypic brain slices, and recording EEG in rodents. B) Have publications and preliminary data to submit a strong R01 application by the 4th year of the award. My final goal is to be an independent researcher in inhibition physiology and pediatric epilepsy, specifically in relation to traumatic brain injury (TBI). TBI is a leading caus of morbidity and mortality in the pediatric age group causing more than 50% of childhood deaths. Seizures are a common complication in pediatric TBI. How TBI leads to seizures is unclear. Several pathological conditions, including brain injury, induce an elevation of the neuronal chloride concentration [Cl-] i. The [Cl-] i am of great importance in neurons as Cl- flux through GABAA receptors mediate inhibition when [Cl-] i is lower than the neuron's resting membrane potential, but can lead to excitation if [Cl-] i is above the resting membrane potential. Our most recent publication has shown that mannitol, and osmotic agent used to treat brain edema in the Neurological Intensive Care Unit, not only decreases the neuronal volume, but also reduces [Cl-]i. Therefore, mannitol should have anti-convulsive properties. My proposal aims to answer the following hypothesis: mannitol (a prototypical hyperosmolar agent) reduces epileptiform activity in the neocortex due to a decrease in [Cl-] i. This decrease in [Cl-] i lead to an increase efficac of GABAAR mediated inhibition in a model of pediatric TBI. To answer this hypothesis, we will use in vitro / in vivo multiphoton imaging and electrophysiological techniques in a transgenic mouse that expresses a new Cl- sensitive radiometric fluorophore (SuperClomeleon). The K08 application, through courses and hands on experience, has been targeted to obtain the necessary skills in: A) multiphoton in vivo imaging; B) experience working with and culturing organotypic brain slices; C) recording and analyzing EEG in rodents. These techniques will allow me to complete my proposed project and give me the required technical knowledge to complement my current in vitro electrophysiology skills. I have a strong background in basic research and clinical pediatric neurology allowing me to understand and see the despair that families go through due to the limitations of post-traumatic epilepsy treatment. Furthermore, my 11 original published articles and my track record of competitive funding with at least one paper delivered per funding agency are evidence to my high motivation, ability to complete projects and my desire to continue a career in research targeted to neuronal inhibitory physiology and epilepsy. I am a full time faculty at Massachusetts General Hospital (MGH) in the Pediatric Neurology division of the Department of Neurology and an Instructor in Neurology at Harvard Medical School. I have the support of the Chair of the Department of Neurology and of the Chief of Pediatric Neurology to become a productive and independent researcher at MGH. With cutting edge technologies and facilities at MGH (2-photon microscopes, electrophysiological equipment), mentors with excellent track records (Dr. Staley, Dr. Bacskai), an advisory committee to discuss my research progress and translational possibilities (Dr. Rosand, Dr. Duhaime, Dr. Cash), the collaborative atmosphere at MGH and being part of Harvard Medical School, I am in a an excellent environment to develop a strong, productive and rewarding career as an independent researcher. The K08 training will be fundamental to help me succeed as an independent investigator.