Canine Model of Late-Infantile Neuronal Ceroid Lipofuscinosis for Therapy Develop

描述（由申请方提供）：神经元蜡样脂褐质沉积症（NCL）是一种遗传性、致死性神经退行性疾病。这些疾病的特征在于进行性神经症状，包括癫痫发作、失明以及认知和运动退化。症状的发作几乎总是发生在儿童时期，通常在6个月至7岁之间，这取决于疾病亚型。在病理学上，NCL的标志是自体荧光溶酶体储存物质在整个中枢神经系统中的积累，伴随着广泛的神经元死亡。至少八种不同基因之一的突变可能导致人类NCL。目前还没有针对任何NCL的有效治疗方法。本提案中概述的研究目标是开发和表征NCL晚期婴儿形式（LINCL）的犬模型，也称为CLN2。一旦开发完成，该模型将用于进行临床前研究，以测试基因疗法、酶替代疗法和间充质干细胞疗法治疗LINCL的安全性和有效性。CLN2是一种隐性疾病，由三肽基肽酶1（TPP1）基因突变引起。TPP1是一种可溶性溶酶体酶，通过细胞内吞途径在细胞之间交换。我们发现腊肠犬在TPP1的犬直系同源基因中有移码突变，并发展出与人类CLN2非常相似的疾病。我们建议彻底描述的表型特征LINCL在一个研究群体的TPP1突变腊肠。这些研究将是临床前研究的前奏，这些研究旨在评估治疗干预的安全性和有效性，以向中枢神经系统提供缺失的酶。体外和体内实验都表明，外源性提供这种酶可以纠正TPP1突变的动物和人类细胞和组织中的生化缺陷。一旦犬模型得到了很好的表征，我们建议在NIH U01资助机制下寻求资金，在该模型中进行临床前研究，以确定是否可以通过直接酶输注、遗传修饰的自体间充质干细胞或基因替代疗法向大脑和视网膜提供功能性TPP1酶来改善疾病表型。 公共卫生相关性：神经元蜡样质脂褐质沉积症（NCL），也称为巴滕病，是一种遗传性进行性神经退行性疾病，几乎总是在儿童期发病，并且总是致命的。已知至少8种不同基因的突变会导致NCL，并且尚未针对这些疾病中的任何一种开发出有效的疗法。使用一种独特的NCL狗模型，我们计划进行临床前研究，以评估这些疾病的有希望的治疗方法。

DESCRIPTION (provided by applicant): Neuronal ceroid lipofuscinoses (NCLs) are inherited, fatal neurodegenerative disorders. These disorders are characterized by progressive neurological symptoms, including seizures, blindness, and cognitive and motor deterioration. Onset of symptoms almost always occurs in childhood, usually between the ages of 6 months and 7 years, depending on the disease subtype. Pathologically, the hallmarks of the NCLs are the accumulation of autofluorescent lysosomal storage material throughout the central nervous system, accompanied by widespread neuronal death. Mutations in one of at least eight different genes can lead to NCL in humans. No effective treatment has yet been developed for any of the NCLs. The goal of the research outlined in this proposal is to develop and characterize a canine model for the late-infantile form of NCL (LINCL), also known as CLN2. Once developed, this model will be used to perform pre- clinical studies to test the safety and efficacy of gene therapy, enzyme replacement therapy and mesenchymal stem cell therapy for treating LINCL. CLN2 is a recessive disorder that results from mutations in the tripeptidyl peptidase 1 (TPP1) gene. TPP1 is a soluble lysosomal enzyme that is exchanged between cells via the cellular endocytic pathway. We discovered Dachshunds that have a frameshift mutation in the canine ortholog of TPP1 and develop a disease very similar to human CLN2. We propose to thoroughly characterize the phenotypic features of LINCL in a research colony of TPP1 mutant Dachshunds. These studies will be a prelude to the conduct of preclinical studies of the safety and efficacy of therapeutic interventions to supply the missing enzyme to the central nervous system. Both in vitro and in vivo experiments have demonstrated that supplying the enzyme exogenously can correct the biochemical defect in cells and tissues of animals and humans with TPP1 mutations. Once the canine model has been well characterized, we propose to seek funding under the NIH U01 funding mechanism to conduct pre-clinical studies in this model to determine whether the disease phenotype can be ameliorated by supplying functional TPP1 enzyme to the brain and retina via direct enzyme infusion, delivery by genetically modified autologous mesenchymal stem cells, or gene replacement therapy. PUBLIC HEALTH RELEVANCE: Neuronal ceroid lipofuscinoses (NCLs), also known as Batten disease, are inherited progressive neurodegenerative disorders that almost always have a childhood onset and are invariably fatal. Mutations in at least 8 different genes are known to cause NCL, and no effective therapies have yet been developed for any of these disorders. Using a unique dog model of one form of NCL, we plan to conduct pre-clinical research to evaluate promising therapies for these diseases.