Gene Therapy for Upper Airway and Respiratory Insufficiency in an ALS Mouse Model

ALS 小鼠模型中上气道和呼吸功能不全的基因治疗

• 批准号: 9581552
• 负责人: Mai ElMallah
• 金额: $ 20.94万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9581552
• 关键词: Gene; Therapy; Upper; Airway; Respiratory

Project Summary/Abstract Amyotrophic Lateral Sclerosis (ALS) is a devastating neurodegenerative disease that initially manifests with either bulbar or spinal (limb) symptoms. Bulbar involvement leads to speech and swallowing impairment and recurrent aspiration pneumonia. Death due to respiratory failure occurs within 2-3 years for patients with bulbar onset ALS and 3-5 years for limb onset cases. To date, no treatment exists for this relentless and fatal disease. The only FDA approved drug for ALS is Riluzole – a glutamate release inhibitor which minimally increases life by 3-5 months. As diaphragm and respiratory muscle weakness progress, targeted respiratory therapy with non-invasive ventilatory support improves survival. Therefore, the goal of this proposal is to use gene therapy to target the respiratory system (upper airway, diaphragm and intercostals) in a popular ALS mouse model – the SOD1G93A mouse and assess if this improves breathing and survival. This application proposes to study the impact of respiratory directed gene therapy on spontaneous breathing, pulmonary mechanics, and respiratory efferent nerve output. Adeno- associated virus (AAV) gene therapy coupled with microRNA that silences SOD1 (miRSOD1) will be delivered through retrograde transduction to the entire motor unit (muscle, neuromuscular junction, nerve and motor neurons). In addition, this proposal aims to study the impact of respiratory targeted and systemic gene therapy on pulmonary physiology and neurophysiology outcome measures as well as behavioral testing and survival. Thus, the fundamental hypothesis driving this proposal is that AAV- miRSOD1 gene therapy targeting the upper airway and respiratory system will enhance breathing in the ALS SOD1G93A murine model; and when coupled with systemic delivery of AAV, will improve respiratory function, mobility and prolong survival. Two specific aims are proposed: Aim 1 will test the hypothesis that an intralingual and intrathoracic injection of AAV-miRSOD1 will effectively transduce the entire hypoglossal and phrenic motor units and stimulate respiratory drive. Aim 2 will test the hypothesis that systemic AAV-miRSOD1 delivery coupled with respiratory targeted therapy will halt degeneration, improve survival and enhance respiratory function. Project Relevance: This work is innovative because it will use gene therapy to target respiratory insufficiency (the main cause of death in ALS patients) in an ALS mouse model. The impact of this therapy will be assessed using translational pulmonary outcome measures with the ultimate goal of translating this therapy to the clinic. AAV gene therapy is now in clinical trials and if successful, this therapy will provide a therapeutic option for respiratory failure in the patients with mutations in the SOD1 gene.

项目摘要/摘要 肌萎缩侧索硬化症是一种毁灭性的神经退行性疾病，最初 有球部或脊椎(四肢)症状的症状。延髓受累导致言语和 吞咽障碍和反复吸入性肺炎。因呼吸衰竭死亡 对于球起病的ALS患者，发生在2-3年内，对于肢体起病的患者，发生在3-5年内。 到目前为止，还没有针对这种无情而致命的疾病的治疗方法。FDA批准的唯一一种药物 治疗肌萎缩侧索硬化症的药物是利鲁唑，这是一种谷氨酸释放抑制剂，可以最小限度地延长生命3-5个月。 随着横隔膜和呼吸肌无力的进展，有针对性的呼吸治疗 无创呼吸机支持可提高存活率。因此，这项提案的目标是使用 靶向呼吸系统(上呼吸道、横隔膜和肋间)的基因治疗 广受欢迎的ALS小鼠模型-SOD1G93A小鼠，并评估这是否改善呼吸和 生死存亡。这项应用建议研究呼吸导向基因治疗对 自主呼吸、肺力学和呼吸传出神经输出。腺体- 相关病毒(AAV)基因治疗结合沉默SOD1的microRNA(MiRSOD1)将 通过逆行转导传递到整个运动单位(肌肉、神经肌肉 连接、神经和运动神经元)。此外，这项建议旨在研究 呼吸靶向全身基因治疗对肺生理和神经生理学的影响 结果测量以及行为测试和存活率。因此，根本的 支持这一建议的假设是，针对上呼吸道的AAV-miRSOD1基因治疗 和呼吸系统将增强ALS SOD1G93A小鼠模型的呼吸；以及何时 再加上全身注射AAV，将改善呼吸功能，活动能力和延长 生死存亡。提出了两个具体的目标：目标1将检验这样一个假设，即语言内和 胸腔内注射AAV-miRSOD1将有效地转导整个舌下神经和 横隔膜运动单位和刺激呼吸驱动。目标2将检验系统性的假设 AAV-miRSOD1载体联合呼吸靶向治疗将阻止退变，改善 活下来，增强呼吸功能。项目相关性：这项工作具有创新性，因为 它将使用基因疗法来治疗呼吸功能不全(ALS的主要死亡原因 患者)在ALS小鼠模型中。这种疗法的影响将使用以下方法进行评估 转化性肺结果测量，最终目标是将这种疗法转化为 诊所。AAV基因疗法目前正在进行临床试验，如果成功，这种疗法将提供一种 SOD1基因突变患者呼吸衰竭的治疗选择。