Control of Breathing and Pompe Disease

呼吸控制和庞贝病

• 批准号: 10152637
• 负责人: BARRY J BYRNE
• 金额: $ 59.32万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10152637
• 关键词: Address; Adult; Age; Animal Model; Antibodies; Brain; Breathing; CRISPR/Cas technology; Cardiac; Cerebrospinal Fluid; Cessation of life; Child; Creatine Kinase; Data; Dependovirus; Desmin; Disease; Disease Progression; Dose; Early treatment; Enzymes; Failure; Gene Mutation; Genes; Glucan 1,4-alpha-Glucosidase; Glycogen; Glycogen storage disease type II; Goals; Grant; Heart failure; Human; Immune; Immune response; Immunosuppression; Impairment; Injections; Intravenous; Late-Onset Disorder; Life; Longevity; MM form creatine kinase; Mediating; Modeling; Mus; Muscle; Mutation; Myocardium; Neonatal; Nerve Degeneration; Nervous system structure; Neuraxis; Neuromuscular Diseases; Neurons; Onset of illness; Pathology; Patients; Phase I/II Clinical Trial; Phenotype; Proteins; Rattus; Recombinants; Residual state; Respiration; Respiratory Diaphragm; Respiratory Failure; Serotyping; Skeletal Muscle; Spinal Cord; Symptoms; Synapsins; Testing; Therapeutic; Tissues; Transgenes; Treatment Efficacy; Work; base; early onset; enzyme replacement therapy; enzyme therapy; gallium arsenide; gene therapy; heart function; heart preservation; life-sustaining therapy; male; mortality; neuron loss; neuropathology; pediatric cardiologist; pre-clinical; preclinical development; preservation; prevent; promoter; rat genome; relating to nervous system; respiratory; response; standard of care; vector

Project Summary / Abstract: Pompe disease results from mutations in the gene for acid α-glucosidase (GAA) – an enzyme necessary to degrade lysosomal glycogen. Early-onset disease occurs in the absence of functional GAA which leads to cardiorespiratory failure early in life. Late-onset disease is associated with reduced GAA activity and gradual progression to respiratory failure. Work from our first two grant cycles indicates neural involvement in respiratory failure in Gaa-/- mice and Pompe patients. This is relevant since the standard of care – intravenous enzyme therapy using recombinant GAA - does not reach the central nervous system (CNS) and patients still progress to respiratory failure. Our overarching hypothesis is that adeno-associated virus (AAV) therapy is capable of restoring life-long GAA expression throughout the CNS, skeletal and cardiac muscle, thereby preserving cardiorespiratory function and prolonging life. Aim 1 focuses on AAV therapy for early-onset disease which requires early life treatments that can prevent both respiratory and cardiac failure. To better study this problem, we created a Gaa null (Gaa-/-) rat model which recapitulates the early onset phenotype with cardiorespiratory pathology and early mortality. Preliminary data indicate that neonatal AAV-GAA therapy (desmin promoter, AAV9 serotype) evokes no detectable immune response, mitigates cardiac and respiratory pathology and prevents early death. Thus, we hypothesize that a single intravenous AAV-GAA dose in young rats can drive persistent and widespread GAA expression and extend the Pompe rat lifespan. Aim 2 addresses late onset Pompe disease in which respiratory failure is the primary cause of mortality. Based on data from our first two grant cycles we hypothesize that neural directed AAV-GAA therapy in adult Pompe rats is sufficient to prevent respiratory decline and extend the lifespan. By packaging AAV-GAA with muscle (creatine kinase 8), neural (synapsin) or tissue specific (muscle and neural, desmin) promoters, and delivering the vector intrathecally, intravenously, or both, we can drive GAA expression in a manner that will determine if neural correction is necessary and sufficient to prevent decline. The aforementioned Gaa null rat will be used to test proof-of-concept for neural vs. muscle correction in the absence of endogenous GAA activity. We will also use another new Pompe rat model in which CRISPR/cas9 has been used to insert the most common human gene mutation causing late-onset Pompe disease (IVS1) into the rat genome. This is important because the IVS1 mutation leads to low but not absent GAA activity and is associated with delayed progression to respiratory failure. The proposed work is significant because current therapeutic strategies in Pompe disease only delay disease progression with eventual respiratory failure. The strategies proposed here will also contribute to the broader goal of advancing gene therapy for neurodegenerative conditions and autosomal recessive diseases.

项目摘要/摘要： 庞贝病是由酸性 α-葡萄糖苷酶 (GAA) 基因突变引起的，GAA 是一种必需的酶。 降解溶酶体糖原。早发性疾病发生在缺乏功能性 GAA 的情况下，从而导致 生命早期出现心肺衰竭。晚发性疾病与 GAA 活性降低和逐渐 进展为呼吸衰竭。我们前两个资助周期的工作表明神经参与 Gaa-/- 小鼠和庞贝氏症患者的呼吸衰竭。这是相关的，因为护理标准 – 静脉注射 使用重组 GAA 的酶疗法 - 无法到达中枢神经系统 (CNS)，患者仍然 进展为呼吸衰竭。我们的首要假设是腺相关病毒（AAV）疗法 能够恢复整个中枢神经系统、骨骼和心肌的终生 GAA 表达，从而 保护心肺功能，延长寿命。目标 1 侧重于早发型 AAV 治疗 需要早期治疗的疾病，以预防呼吸衰竭和心力衰竭。为了更好 为了研究这个问题，我们创建了 Gaa null (Gaa-/-) 大鼠模型，该模型概括了早发表型 心肺病理学和早期死亡率。初步数据表明新生儿 AAV-GAA 治疗 （结蛋白启动子，AAV9 血清型）不会引起可检测到的免疫反应，减轻心脏和呼吸系统的反应 病理并防止过早死亡。因此，我们假设单次静脉注射 AAV-GAA 剂量对年轻人来说 大鼠可以驱动持久且广泛的 GAA 表达并延长庞贝大鼠的寿命。目标2 解决迟发性庞贝病，其中呼吸衰竭是死亡的主要原因。基于 根据我们前两个资助周期的数据，我们假设对成年 Pompe 大鼠进行神经定向 AAV-GAA 治疗 足以防止呼吸衰退并延长寿命。通过将 AAV-GAA 与肌肉包装在一起 （肌酸激酶 8）、神经（突触蛋白）或组织特异性（肌肉和神经、结蛋白）启动子，以及递送 通过鞘内、静脉内或两者，我们可以驱动 GAA 表达，以确定是否 神经矫正对于防止衰退是必要且充分的。将使用前述的Gaa无效大鼠 测试在缺乏内源性 GAA 活性的情况下神经与肌肉校正的概念验证。我们将 还使用另一种新的 Pompe 大鼠模型，其中使用 CRISPR/cas9 插入最常见的 导致迟发性庞贝病（IVS1）的人类基因突变进入大鼠基因组。这很重要 因为 IVS1 突变导致 GAA 活性低但并非缺失，并且与延迟相关 进展为呼吸衰竭。拟议的工作意义重大，因为当前的治疗策略 庞贝病只会延迟疾病进展，最终导致呼吸衰竭。这里提出的策略 还将有助于推进神经退行性疾病基因治疗的更广泛目标， 常染色体隐性遗传疾病。