CHARACTERIZATION OF FELINE NIEMANN-PICK TYPE C

描述(改编自申请人的摘要)：最初的目标 这个项目是为了开发、表征和维护人类的猫科动物模型 向其他调查人员提供鼻咽癌组织和小猫，并 评估各种治疗方式。本次大赛的总体目标是 更新本质上是相同的。这个项目的具体目标是：1) 继续维持这个猫科鼻咽癌群体，继续做这个模式 可供其他研究人员使用，2)进一步描述猫科动物模型 通过使用针对猫NPC1蛋白产生的抗体，以及3) 继续评估一种神经节苷脂合成抑制剂作为治疗鼻咽癌的方法， 因为一项初步研究表明，治疗后猫的神经节苷脂储存减少 使用这种抑制剂。鼻咽癌在人类和猫的模型中都是 常染色体隐性遗传性溶酶体储存障碍 可能的特征包括器官肿大、进行性神经异常 与神经节苷脂储存和过早死亡有关。初级阶段 细胞学缺陷与细胞内转运缺陷有关 未酯化的胆固醇。NPC1蛋白现在被认为也起到了作用 在神经节苷脂运输中。导致最常见类型的 人鼻咽癌(NPC1)已被鉴定。利用CAT和AND进行互补研究 人鼻咽癌成纤维细胞支持猫鼻咽癌相关基因 该模型与人类NPC1基因同源。猫科动物特异的聚合酶链式反应 生成并设计为放大六个重叠的频段，这些频段跨越整个 猫科鼻咽癌开放阅读框架。预测的猫科动物氨基酸序列显示 与人类NPC1蛋白有91%的同源性。单个碱基替换，结果 在保守的半胱氨酸到丝氨酸的替代中，已经在所有鼻咽癌中发现 受影响的猫进行了评估，并在同一区域内的几个点 在人类中报告的突变。携带者是相同的杂合子 等位基因。已经开发出一种基于聚合酶链式反应的快速准确的检测方法 识别携带者以及受影响的小猫。该模型将提供一种 独特而有价值的资源，用于评估治疗方式 全国人大。现在，受鼻咽癌影响的小猫数量将会增加，因为 携带者检测消除了耗时的育种试验。大约12个 CSU将保留女性和三名男性携带者，目标是 每年生产15到20只受影响的小猫。额外的饲养员和 受影响的小猫将提供给其他调查人员。

DESCRIPTION (Adapted from the applicant's abstract): The original goal of this project was to develop, characterize and maintain a feline model of human NPC, to provide NPC tissues and kittens to other investigators, and to evaluate various treatment modalities. The overall goal of this Competitive Renewal is essentially the same. The specific aims of this project are 1) to continue to maintain this colony of feline NPC and continue to make this model available to other investigators, 2) to further characterize the feline model by using antibodies generated against the feline NPC1 protein, and 3) to continue to evaluate a ganglioside synthesis inhibitor as therapy for NPC, because a pilot study has shown decreased ganglioside storage in cats treated with this inhibitor. NPC in human beings as well as in the feline model, is an autosomal recessive lysosomal storage disorder with a variety of clinical features including organomegaly, progressive neurologic abnormalities possibly related to neuronal ganglioside storage, and premature death. The primary cytologic defect is associated with defective intracellular transport of unesterified cholesterol. The NPC1 protein is now thought to also play a role in ganglioside transport. The gene responsible for the most common type of human NPC (NPC1) has been identified. Complementation studies using cat and human NPC fibroblasts support that the gene responsible for NPC in the feline model is orthologus to human NPC1. Feline specific PCR primers were generated and designed to amplify six overlapping bands which span the entire feline NPC open reading frame. The predicted feline amino acid sequence shows 91% homology to the human NPC1 protein. A single base substitution, resulting in a conserved cysteine to serine substitution, has been identified in all NPC affected cats evaluated, and is in the same area as several of the point mutations reported in human beings. Carriers are heterozygous for the same allele. A PCR-based assay has been developed that quickly and accurately identifies carriers, as well as affected kittens. This model will provide a unique and valuable resource with which to evaluate treatment modalities for NPC. Increased numbers of NPC-affected kittens will now be available, since carrier detection eliminates time-consuming breeding trials. Approximately 12 female and three male carriers will be maintained at CSU, with the goal of producing 15 to 20 affected kittens per year. Additional breeders and affected kittens will be made available to other investigators.