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.

项目摘要/摘要 神经元粘膜脂蛋白（NCLS或Batten疾病）是最具破坏性的遗传性疾病之一 童年和美国儿童神经退行性的最常见原因目前没有 治愈这些疾病，并且治疗基本上仍然支持。 NCL的特征是渐进式 未基因的细胞材料的内溶性积累；这种积累被认为是由于缺陷而产生的 在自噬 - 溶酶体途径中，但本身可能有助于发病机理。我们的数据表明缺陷 在少年棕褐色疾病蛋白中，CLN3损害了一部分溶酶体酶的成熟，并损害 TFEB的天山糖介导的激活，TFEB是自噬 - 溶糖途径的主要调节剂，可以改善 幼体少年芭蕾疾病（JNCL）的疾病负担。我们建议研究新型机制 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率缺陷引起。首先，我们将测试海藻糖 - 诱导的溶酶体增强校正JNCL小鼠中溶酶体酶的成熟有缺陷（AIM 1）。 我们将通过进行蛋白质成熟和无偏蛋白质组学的实验来检验这一假设 根据使用敲入lamp1flag小鼠系的使用分析，我们已经生成有效隔离 小鼠组织的溶酶体。其次，我们将检验以下假设，即Akt（激酶）的降低或抑制 我们已经确定的TFEB抑制剂将减少JNCL小鼠的神经病理学（AIM 2）。我们将减少AKT 通过使用两种互补方法的活动：从遗传上，通过使用Akt1 - / - 小鼠，并通过药理学，通过 使用目前正在临床发育中的AKT药物抑制剂。第三，我们将检验以下假设 通过调节两种正交途径的TFEB协同药理激活将导致 自噬溶质体系统的更大增强和JNCL病理标志的更好降低 比单独的策略（目标3）。该假设是基于我们的发现，即非受体酪氨酸 激酶SRC是激活MTORC1的重要因素，MTORC1是TFEB的另一种激酶抑制剂。这些研究 在神经退行性疾病模型中，TFEB的药理学会激活。如果成功，这 研究将提供强大的TFEB激活范式，这可能为临床治疗奠定基础 棕褐色疾病，并有可能由损害损害引起的其他神经退行性储存障碍 自噬溶质体途径。