Modulation of lysosomal function for the treatment of Batten disease

调节溶酶体功能治疗巴顿病

• 批准号: 9976592
• 负责人: Marco Sardiello
• 金额: --
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2020-08-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9976592
• 关键词: AKT inhibition; Adolescent; Autophagocytosis; Basic Science; Binding; Biogenesis; Biological Assay; Brain; CLN3 gene; Carbohydrates; Cells; Cessation of life; Chemicals; Child; Childhood; Clinical Treatment; Dasatinib; Data; Defect; Dementia; Deterioration; Disease; Disease Progression; Enhancers; Enzymes; Estrogen receptor positive; Event; FDA approved; Foundations; Functional disorder; Genetic; Goals; Golgi Apparatus; Guanosine Triphosphate Phosphohydrolases; Hereditary Disease; IGF Type 2 Receptor; Impairment; Inherited; Knock-in; Lead; Lysosomes; Malignant Neoplasms; Mediating; Modeling; Monitor; Motor; Mus; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Ceroid-Lipofuscinosis; Neurons; Onset of illness; Pathogenesis; Pathologic; Pathology; Pathway interactions; Pharmaceutical Preparations; Pharmacology; Phase; Phenotype; Protein-Serine-Threonine Kinases; Proteins; Proteomics; Regimen; Regulatory Pathway; Resolution; Role; Route; Sirolimus; Spielmeyer-Vogt Disease; System; Testing; Therapeutic Intervention; Tissues; Translating; Trehalose; Work; base; burden of illness; clinical development; combinatorial; dosage; experimental study; in vivo; inhibitor/antagonist; kinase inhibitor; lysosomal proteins; mannose 6 phosphate; mouse model; neuroinflammation; neuron loss; neuropathology; novel; novel therapeutics; prevent; protein kinase inhibitor; recruit; side effect; small molecule inhibitor; therapy development; trafficking

PROJECT SUMMARY/ABSTRACT Neuronal ceroid lipofuscinoses (NCLs or Batten disease) are among the most devastating inherited disorders of childhood and the most common cause of neurodegeneration in children in the U.S. There is currently no cure for these disorders, and treatments remain largely supportive. NCLs are characterized by the progressive intralysosomal accumulation of undegraded cellular material; this accumulation is thought to result from defects in the autophagy-lysosomal pathway, but could itself contribute to pathogenesis. Our data show that deficiency of the juvenile Batten disease protein, CLN3, impairs maturation of a subset of lysosomal enzymes and that trehalose-mediated activation of TFEB, a master regulator of the autophagy-lysosomal pathway, ameliorates disease burden in a mouse model of juvenile Batten disease (JNCL). We propose to study novel mechanisms of TFEB activation that could lead to translational applications for JNCL and other neurodegenerative disorders caused by defects in lysosome-mediated cellular clearance. First, we will test the hypothesis that trehalose- induced lysosomal enhancement corrects defective maturation of lysosomal enzymes in JNCL mice (Aim 1). We will test this hypothesis by conducting experiments of protein maturation and by unbiased proteomic analyses based on the use of a knock-in Lamp1FLAG mouse line we have generated to efficiently isolate lysosomes from mouse tissues. Second, we will test the hypothesis that reduction or inhibition of Akt, a kinase inhibitor of TFEB we have identified, will decrease neuropathology of JNCL mice (Aim 2). We will reduce Akt activity by using two complementary approaches: genetically, by using Akt1-/- mice, and pharmacologically, by using an Akt drug inhibitor that is currently in clinical development. Third, we will test the hypothesis that synergistic pharmacological activation of TFEB by modulation of two orthogonal pathways will result in a greater enhancement of the autophagy-lysosomal system and better reduction of JNCL pathological hallmarks than either strategy alone (Aim 3). This hypothesis is based on our finding that the non-receptor tyrosine kinase, Src, is an essential factor for activation of mTORC1, another kinase inhibitor of TFEB. These studies will pioneer pharmacological activation of TFEB in a model of neurodegenerative disorder. If successful, this study will provide a powerful paradigm of TFEB activation that could lay the foundation for the clinical treatment of Batten disease and, potentially, additional neurodegenerative storage disorders caused by impairment of the autophagy-lysosomal pathway.

项目概要/摘要 神经元蜡样脂褐质沉积症（NCL 或 Batten 病）是最具破坏性的遗传性疾病之一 童年时期的疾病，也是美国儿童神经退行性变的最常见原因。目前还没有 治愈这些疾病，并且治疗在很大程度上仍然是支持性的。 NCL 的特点是进行性 未降解的细胞物质在溶酶体内积累；这种积累被认为是缺陷造成的 在自噬-溶酶体途径中，但其本身可能有助于发病机制。我们的数据表明，缺乏 幼年巴顿病蛋白 CLN3 会损害溶酶体酶子集的成熟 海藻糖介导的 TFEB 激活（自噬溶酶体途径的主要调节因子）可改善 幼年巴顿病（JNCL）小鼠模型的疾病负担。我们建议研究新机制 TFEB 激活可能导致 JNCL 和其他神经退行性疾病的转化应用 由溶酶体介导的细胞清除缺陷引起。首先，我们将检验以下假设：海藻糖- 诱导的溶酶体增强可纠正 JNCL 小鼠溶酶体酶的成熟缺陷（目标 1）。 我们将通过进行蛋白质成熟实验和无偏蛋白质组学实验来检验这一假设 基于使用我们生成的敲入 Lamp1FLAG 小鼠品系进行分析，以有效分离 来自小鼠组织的溶酶体。其次，我们将检验以下假设：Akt（一种激酶）的减少或抑制 我们已经确定的 TFEB 抑制剂将减少 JNCL 小鼠的神经病理学（目标 2）。我们将减少 Akt 活性通过使用两种互补的方法：遗传上，通过使用 Akt1-/- 小鼠，以及药理学上，通过 使用目前处于临床开发阶段的 Akt 药物抑制剂。第三，我们将检验以下假设： 通过调节两个正交途径对 TFEB 进行协同药理学激活将导致 自噬-溶酶体系统的更大增强和 JNCL 病理标志的更好减少 比单独使用任一策略（目标 3）更有效。该假设基于我们的发现，即非受体酪氨酸 激酶 Src 是激活 mTORC1（另一种 TFEB 激酶抑制剂）的重要因子。这些研究 将在神经退行性疾病模型中开创 TFEB 的药理学激活。如果成功的话，这 研究将为 TFEB 激活提供强大的范例，为临床治疗奠定基础 Batten 病，以及可能由神经功能受损引起的其他神经退行性储存障碍 自噬-溶酶体途径。