Novel Approaches for Correcting Respiratory Insufficiency in Pompe Disease

描述（由申请人提供）：El Mallah博士是佛罗里达大学儿科肺科儿科助理教授，75%的时间用于研究，25%用于临床。在拟议的奖励期间，她的职业发展和学习目标是1）获得呼吸神经生物学和分子治疗的核心知识，2）获得研究方法，技术和科学写作的进一步专业知识，以及3）发展学术领导技能。El Mallah博士的长期职业目标是成为一名成功的独立儿科肺部临床科学家，重点是转化研究。由于她的研究，她发表了她的研究结果，在全国会议上提出，并获得了几个奖项和赠款，包括帕克B。弗朗西斯奖学金。环境：富勒博士和伯恩博士是杰出的导师，有成功的学员记录。富勒博士是呼吸神经生物学的领导者，而伯恩博士是国家基因治疗专家。他们都有大量的NIH资金，非常活跃和富有成效的实验室，并将提供资源，以协助候选人在她的研究项目。佛罗里达大学（UF）儿科提供了一个结构化的学者计划，旨在提高研究和领导能力，并给予结构化的反馈和评估。UF还提供员工教育计划，以支持候选人的课程。研究：本提案中的研究项目将建立在候选人先前针对庞贝氏症呼吸功能障碍的研究基础上。此外，它将探索一种新的治疗剂，用于刺激Pompe（Gaa-/-）小鼠模型的呼吸驱动。庞贝氏症是一种致命的神经肌肉疾病，由酸性α-葡萄糖苷酶（GAA）基因突变引起，GAA是降解溶酶体糖原所必需的酶。患有庞贝氏症的婴儿患有呼吸功能不全，通常导致机械通气。呼吸问题最初被归因于肌肉病理学，但我们的团队最近显示了中枢神经系统的贡献。不幸的是，唯一FDA批准的庞贝氏症疗法（酶替代疗法）是次优的，因为它不能穿过血脑屏障。因此，许多接受酶替代疗法的患者仍然需要机械通气。我们提出了一系列的临床前研究，直接解决的需要治疗的中枢神经系统庞贝氏症。具体来说，我们将评估ampakines的治疗效果，化合物，增强兴奋性多巴胺能神经传递呼吸神经元。此外，由于ampakines不能直接解决潜在的病理学，我们将使用基因递送来纠正潜在的基因缺陷。单独地，基因递送并不覆盖整个运动神经元池，并且似乎仅部分地纠正神经功能障碍。因此，我们建议研究ampakines对Pompe小鼠AAV基因治疗后呼吸功能的影响，以优化呼吸功能障碍的治疗。总的来说，这个项目将测试一个重要的和新颖的假设，并将为候选人提供新的实验技术的培训。驱动这一提议的基本假设是，安巴金治疗将增强庞贝氏症中的呼吸运动输出，并且在CNS GAA活性通过基于AAV的基因治疗增加后，这些药物的功能将进一步增强。这将使用两个特定的目的进行检查：目的1：为了测试安巴金治疗将刺激庞贝氏症小鼠模型中的呼吸驱动的假设。目标二：为了检验通过基于AAV的基因治疗增加CNS GAA活性后安巴金治疗的疗效将进一步增强的假设。项目相关性：这项工作是创新的，因为这些兴奋剂，ampakines对呼吸功能障碍的影响尚未在神经肌肉疾病的背景下进行评估。此外，通过神经机制明确发挥作用的安巴金治疗将有助于强调神经对庞贝氏症小鼠呼吸功能障碍的贡献，以及靶向CNS以治疗这种致命疾病的必要性。此外，由于ampakines和AAV-GAA都已经进行了I期和II期临床试验，并且对人类使用是安全的，因此这项工作具有快速转化为人类护理并影响庞贝氏症临床治疗的强大潜力。

DESCRIPTION (provided by applicant): Dr. El Mallah is an Assistant Professor of Pediatrics, Pediatric Pulmonary Division, University of Florida, with 75% of time devoted to research, 25% to clinical. During the proposed award, her career development and learning objectives are to 1) acquire core knowledge in respiratory neurobiology and molecular therapy, 2) gain further expertise in research methodology, techniques, and scientific writing, and 3) develop academic leadership skills. Dr. El Mallah's long-term career goal is to become a successful independent pediatric pulmonary clinician scientist with an emphasis on translational research. As a result of her research she has published her findings, presented them at national meetings and has received several awards and grants, including the Parker B. Francis fellowship award. ENVIRONMENT: Drs. Fuller and Byrne are exceptional mentors, with a track record of successful mentees. Dr. Fuller is a leader in respiratory neurobiology, while Dr. Byrne is a national gene therapy expert. They both have extensive NIH funding, very active and productive laboratories, and will provide the resources to assist the candidate in her research project. The University of Florida (UF) Pediatric Department offers a structured scholars program designed to enhance research and leadership skills, and to give structured feedback and evaluations. UF also offers an employee education program to support coursework for the candidate. RESEARCH: The research project in this proposal will build on the candidate's prior research on targeting respiratory dysfunction in Pompe disease. In addition, it will explore a novel therapeutic agent for stimulating respiratory drive in a Pompe (Gaa-/-) mouse model. Pompe disease is a fatal neuromuscular disorder resulting from mutations in the gene for acid alpha- glucosidase (GAA) - an enzyme necessary to degrade lysosomal glycogen. Infants with Pompe disease suffer with respiratory insufficiency often leading to mechanical ventilation. Breathing problems have originally been attributed to muscle pathology but our group has recently shown a central nervous system contribution. Unfortunately, the only FDA approved therapy for Pompe disease (enzyme replacement therapy) is suboptimal because it does not cross the blood brain barrier. Therefore, many patients on enzyme replacement therapy still require mechanical ventilation. We propose a series of pre-clinical studies that directly address the need for treatment of the CNS in Pompe disease. Specifically, we will evaluate the therapeutic efficacy of ampakines, compounds that enhance excitatory glutaminergic neural transmission in respiratory neurons. In addition, since ampakines do not directly address the underlying pathology, we will use gene delivery to correct the underlying gene defect. Alone, gene delivery does not transduce the entire motoneuron pool and appears to only partially correct neural dysfunction. Therefore we propose to study the impact of ampakines on respiratory function following AAV gene therapy in Pompe mice in order to optimize therapy for respiratory dysfunction. Overall, this project will test an important and novel hypothesis and will provide the candidate with training in new experimental techniques. The fundamental hypothesis driving this proposal is that ampakine therapy will potentiate respiratory motor output in Pompe disease, and the function of these drugs will be further enhanced after CNS GAA activity is increased via AAV-based gene therapy. This will be examined using two specific aims: Aim 1: To test the hypothesis that ampakine therapy will stimulate respiratory drive in a mouse model of Pompe disease. Aim 2: To test the hypothesis that the efficacy of ampakine therapy will be further enhanced after CNS GAA activity is increased via AAV-based gene therapy. PROJECT RELEVANCE: This work is innovative because the impact of these excitatory agents, ampakines on respiratory dysfunction has not been evaluated in the context of neuromuscular disorders. Moreover, the ampakine treatments, which definitively work via a neural mechanism, will serve to emphasize the neural contribution to respiratory dysfunction in Pompe mice, and the necessity to target the CNS in order to treat this fatal disease. In addition, since both ampakines and AAV-GAA have already undergone phase I and II clinical trials and are safe for human use, this work has the strong potential to quickly translate to human care and to impact the clinical treatment of Pompe disease.