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.

项目总结/摘要 神经元蜡样质脂褐质病（NCL或Batten病）是最具破坏性的遗传性疾病之一 在美国，儿童神经退行性疾病的最常见原因。 治疗这些疾病，治疗仍然在很大程度上支持。NCL的特点是渐进的 未降解的细胞物质在溶酶体内的积累;这种积累被认为是由缺陷引起的 自噬-溶酶体途径，但本身可能有助于发病机制。我们的数据显示， 幼年巴滕病蛋白CLN 3的表达损害了溶酶体酶亚群的成熟， 海藻糖介导的TFEB激活，自噬-溶酶体途径的主要调节剂，改善 在幼年巴滕病（JNCL）小鼠模型中的疾病负担。我们建议研究新的机制 TFEB激活可能导致JNCL和其他神经退行性疾病的翻译应用 由溶酶体介导的细胞清除缺陷引起。首先，我们将检验海藻糖- 诱导的溶酶体增强纠正了JNCL小鼠中溶酶体酶的成熟缺陷（Aim 1）。 我们将通过进行蛋白质成熟实验和无偏蛋白质组学方法来验证这一假设。 基于使用敲入Lamp 1FLAG小鼠系的分析，我们已经产生了有效分离 来自小鼠组织的溶酶体。第二，我们将检验Akt激酶的减少或抑制 我们已经鉴定的TFEB的抑制剂将减轻JNCL小鼠的神经病理学（目的2）。我们将减少Akt 使用两种互补方法来提高活性：遗传学上，使用Akt 1-/-小鼠，药理学上，通过 使用目前正在临床开发中的Akt药物抑制剂。第三，我们将检验假设， 通过调节两个正交途径协同药理学激活TFEB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 而不是单独使用任何一种策略（目标3）。这一假设是基于我们的发现，非受体酪氨酸 激酶Src是激活TFEB的另一种激酶抑制剂mTORC 1的必需因子。这些研究 将开创TFEB在神经退行性疾病模型中的药理学激活。如果成功，这 本研究将为TFEB激活提供一个强有力的范例，为临床治疗奠定基础 Batten病，以及潜在的其他神经退行性储存障碍， 自噬-溶酶体途径