Investigating cell-type-specific contribution to JNCL

研究细胞类型特异性对 JNCL 的贡献

• 批准号: 8729039
• 负责人: Beverly L. Davidson
• 金额: $ 16.58万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8729039
• 关键词: Adolescent; Age; Antibodies; Behavioral; Blindness; Blood; Blood - brain barrier anatomy; Blood Vessels; Brain; CLN3 gene; Cell Line; Cells; Ceroid; Cessation of life; Child; Childhood; Cognitive; Complex; Defect; Deterioration; Development; Disease; Disease Progression; Drug Efflux; Endocytosis; Endothelial Cells; Endothelium; Exons; Family; Functional disorder; Galactosidase; Gene Transfer; Genes; Health; Human; Impairment; In Vitro; Inherited; Injection of therapeutic agent; Integral Membrane Protein; LacZ Genes; Maintenance; Measures; Mediating; Molecular; Motor; Mus; Mutation; Nervous system structure; Neuraxis; Neurodegenerative Disorders; Neuroglia; Neurologic; Neuronal Ceroid-Lipofuscinosis; Neuronal Differentiation; Neurons; Nuclear; Nutrient; Outcome; Pathogenesis; Pathology; Pathway interactions; Patients; Pattern; Peptides; Play; Poison; Population; Quality of life; Reading; Reporter; Retina; Role; Route; Seizures; Spielmeyer-Vogt Disease; Symptoms; Technology; Terminator Codon; Testing; Transcript; Transgenic Mice; Translating; Translations; Treatment Efficacy; Tropism; Vascular Endothelial Cell; Visual; adeno-associated viral vector; base; cell type; cellular targeting; disease phenotype; effective therapy; efficacy testing; gene therapy; gene transfer vector; improved; in vivo; loss of function; mouse model; neuron loss; operation; postnatal; prevent; public health relevance; response; restoration; skills; success; therapeutic target; vector

ABSTRACT JNCL is a devastating childhood-onset neurodegenerative disease caused by deficiency in CLN3, a membrane-integral protein with unresolved function. Using a Cln3-reporter mouse, we previously discovered that in the postnatal mouse brain, expression is limited to a few subpopulations of neurons, but is widespread in endothelial cells (EC) that line the vasculature. Our continued studies show that brain EC from CLN3-deficient mice display impairments in drug efflux, regulatory volume response, and endocytosis. These functions are critical to normal operation of the blood-brain barrier (BBB), which governs selective passage of molecules between blood and brain, mediating import of nutrients and export of toxic substances away from proximal neurons. We postulate that brain EC dysfunction plays a dominant role in JNCL pathogenesis, and that restoring CLN3 to EC will alleviate disease progression. The current proposal seeks to test our hypothesis via two main Aims. For Aim 1 we plan to use a transgenic mouse approach to trigger EC-exclusive expression of CLN3, and assess whether this prevents the development of behavioral and pathological measures of JNCL. In Aim 2, we plan to generate an adeno-associated virus (AAV) vector with tropism for brain endothelium to test the efficacy of CLN3 gene transfer in the JNCL mouse model. Accomplishment of these aims will provide valuable information relevant to JNCL pathogenesis and potential treatment avenues. Should our hypothesis prove correct, this would distinguish central nervous system EC as a therapeutic target for JNCL. Moreover, therapeutic efficacy of our gene transfer approach, in which the vector is administered via intra-vascular injection, would be an exciting outcome with hope for translation to JNCL patients, and possibly more broadly to other neurodegenerative diseases.

抽象的 JNCL是由CLN3缺乏引起的毁灭性儿童期神经退行性疾病 具有未分辨功能的膜综合蛋白。使用Cln3-Reporter鼠标，我们以前 发现在产后小鼠大脑中，表达仅限于神经元的一些亚群，但 在脉管系统的内皮细胞（EC）中广泛存在。我们的持续研究表明大脑EC 来自CLN3缺乏的小鼠在药物外，调节体积反应和 内吞作用。这些功能对于血脑屏障（BBB）的正常运行至关重要， 控制分子在血液和大脑之间的选择性通过，介导营养的进口并出口 远离近端神经元的有毒物质。我们假设大脑EC功能障碍起着主导性 在JNCL发病机理中的作用以及将CLN3恢复为EC的作用将减轻疾病的进展。电流 提案旨在通过两个主要目的检验我们的假设。对于目标1，我们计划使用转基因鼠标 触发CLN3表达的EC的方法，并评估这是否阻止了 JNCL的行为和病理度量。在AIM 2中，我们计划生成与腺相关的病毒 （AAV）脑内皮的向量载体测试CLN3基因转移在JNCL中的功效 鼠标模型。这些目标的完成将提供与JNCL相关的有价值的信息 发病机理和潜在的治疗途径。如果我们的假设是正确的，这将区分 中枢神经系统EC作为JNCL的治疗靶标。此外，我们基因的治疗功效 通过血管内注射对矢量进行管理的转移方法将是令人兴奋的 希望转化为JNCL患者的希望，并可能更广泛地转换为其他神经退行性。 疾病。