Pathogenesis and possible therapies modelled in ovine Batten disease

神经元蜡样质脂褐质沉积症（NCLs，Batten病）是一种毁灭性的遗传性疾病 儿童神经退行性疾病目前还没有有效的治疗方法，姑息治疗是 难对新西兰绵羊中CLN6形式的研究已经证明在理解 生物化学和临床前病理学。这些羊有着类似人类的大脑解剖学，生理学和 遗传组织，并显示类似的神经病理学为主的皮质萎缩，以及类似的 临床疾病的发展。这个项目的总体目标是完成对一个点的理解 其中病毒载体基因疗法的体内试验可以开始。分子生物学、生物化学和 免疫组织学实验将确定校正所需的基因产物及其亚细胞位点， 住所将用慢病毒和腺相关病毒衍生的载体测试神经元培养物， 确定将该基因转导入绵羊神经元和神经胶质细胞的适合性。 产前和临床前神经病理学研究表明，神经胶质细胞活化在脑缺血中起主要作用。 本病的发病机制。进一步的研究将确定神经胶质细胞活化和炎症的级联反应， 以确定关键步骤，从而确定最有效的干预点。有效性 将测试用抗炎药物慢性治疗对绵羊疾病发展的影响 in vivo.脑室下区神经发生和细胞向受累区迁移的延长 在病理学研究中也指出了受影响绵羊的大脑。这一程度将是 通过BrdU标记体内新细胞和抗体检测建立。最后， 将病毒载体中的基因携带到需要它的区域的迁移将由靶向的 将携带报告基因的优选载体注射到受影响的羊脑中， 基因表达的扩散。 这些研究旨在提供直接在绵羊上测试基因治疗的所有要求， 与抑制炎症相结合。目标细胞将已被沿着 CLN6基因产物。治疗的时间窗将与角色一起沿着定义 抑制炎症以减缓疾病的进程。最有效的病毒载体 已确定并建立了SVZ注射技术。 REVELANCE：该项目旨在获得知识，以定义大型动物形式的现实疗法， 巴滕病可以测试可能的人类使用。获得的大部分知识将与以下方面有关： 其他疾病和其他治疗策略，如阿尔茨海默病和干细胞移植。

The neuronal ceroid lipofuscinoses (NCLs, Batten disease) are devastating inherited fatal neurodegenerative diseases of children. At present there are no effective therapies and palliative care is difficult. Studies on a CLN6 form in New Zealand sheep have proved invaluable in understanding the biochemistry and preclinical pathology. These sheep have a human-like brain anatomy, physiology and genetic organization, and display similar neuropathology dominated by cortical atrophy, and a similar development of clinical disease. The overall aim of this project is to complete understanding to a point where in vivo trials of viral vector gene therapies can begin. Molecular biology, biochemistry and immunohistological experiments will define the gene product required for correction and its subcellular site of residence. Neuron cultures will be be tested with lenti- and adeno-associated virally derived vectors to determine suitability for transduction of the gene into sheep neurons and glial cells. Prenatal and preclinical neuropathology studies indicated that glial activation has a primary role in the pathogenesis of this disease. Further studies will define the cascade of glial cell activation and inflammation, to determine the critical steps and thus determine the most effective point for intervention. The effectiveness of chronic treatment with anti-inflammatory drugs on the development of the disease in sheep will be tested in vivo. Prolonged neurogenesis and migration of cells from the subventricular zone to the affected areas in the brains of affected sheep was also indicated in the pathology studies. The extent of this will be established by BrdU labeling of new cells in vivo and antibody detection. Finally the usefulness of this migration for carrying a gene in viral vectors to the areas where it is required will be determined by targeted injection of the preferred vector carrying a reporter gene into affected sheep brains and detection of the dispersal of gene expression. These studies are aimed at providing all the requirements for directly testing gene therapy on the sheep, in combination with suppression of inflammation. The target cells will have been identified along with the CLN6 gene product relevant to them. The time window for therapy will have been defined along with the role of suppression of inflammation in slowing the course of the disease. The most effective viral vector will have been identified and the techniques for injection into the SVZ established. REVELANCE: This project intends to gain knowledge to define realistic therapies in a large animal form of Batten disease that can be tested for possible human use. Much of the knowledge gained will be relevant to other diseases and other treatment strategies, such as Alzheimer's disease and stem cell transplantation.