Investigating cell-type-specific contribution to JNCL

摘要 JNCL是一种毁灭性的儿童期起病的神经退行性疾病，由CLN3缺乏引起。 膜整合蛋白，功能未解。使用Cln3报告鼠标，我们之前 发现在出生后的小鼠大脑中，表达仅限于少数神经元亚群，但 广泛存在于血管内皮细胞(EC)中。我们的持续研究表明，大脑EC CLN3缺陷小鼠表现出药物外排、调节性容量反应和 内吞作用。这些功能对血脑屏障(BBB)的正常运行至关重要，血脑屏障 控制血液和大脑之间分子的选择性通道，调节营养物质的输入和输出 远离近端神经元的有毒物质。我们假设脑内皮细胞功能障碍起主导作用 在JNCL的发病机制中发挥作用，将CLN3恢复为EC将减缓疾病的进展。海流 该提案试图通过两个主要目标来检验我们的假设。对于目标1，我们计划使用转基因小鼠 触发CLN3的EC独占表达的方法，并评估这是否阻止了 JNCL的行为学和病理学检查。在目标2中，我们计划产生一种腺相关病毒 脑内皮细胞趋向性(AAV)载体检测CLN3基因转移在JNCL中的作用 老鼠模型。这些目标的实现将为JNCL提供有价值的信息 发病机制和潜在的治疗途径。如果我们的假设被证明是正确的，这将区别于 中枢神经系统EC作为JNCL的治疗靶点。此外，我们的基因的治疗效果 通过血管内注射给予载体的转移方法将是一种令人兴奋的方法 结果有望转化为JNCL患者，并可能更广泛地应用于其他神经退行性变患者 疾病。

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.