Best Hannah L, Neverman Nicole J, Wicky Hollie E, Mitchell Nadia L, Leitch Beulah, Hughes Stephanie M
Department of Biochemistry, School of Biomedical Sciences, Brain Health Research Centre, University of Otago, Dunedin, New Zealand; Batten Animal Research Network (BARN), New Zealand.
Faculty of Agriculture and Life Sciences, Lincoln University, Canterbury, New Zealand; Batten Animal Research Network (BARN), New Zealand.
Neurobiol Dis. 2017 Apr;100:62-74. doi: 10.1016/j.nbd.2017.01.001. Epub 2017 Jan 5.
Batten disease (neuronal ceroid lipofuscinosis) refers to a group of neurodegenerative lysosomal storage diseases predominantly affecting children. There are currently no effective treatments, and the functions of many of the associated gene products are unknown. Here we characterise fetal neural cultures from two genetically distinct sheep forms of Batten disease, with mutations in the lysosomal protein encoding gene CLN5 and endoplasmic reticulum membrane protein encoding gene CLN6, respectively. We found similar reductions in autophagy, acidic organelles and synaptic recycling in both forms compared to unaffected cells. We then developed a high-throughput screen and tested for correction of deficient cells with lentiviral-mediated CLN5 or CLN6 gene transfer and fibrate drugs, gemfibrozil and fenofibrate in CLN6 deficient neural cultures. These assays provide a simple system to rapidly screen candidate therapies or libraries of drugs prior to in vivo testing.
巴顿病（神经元蜡样脂褐质沉积症）是指一组主要影响儿童的神经退行性溶酶体贮积病。目前尚无有效治疗方法，许多相关基因产物的功能也未知。在此，我们对来自两种基因不同的巴顿病绵羊模型的胎儿神经培养物进行了表征，这两种模型分别在溶酶体蛋白编码基因CLN5和内质网内膜蛋白编码基因CLN6中发生了突变。我们发现，与未受影响的细胞相比，两种模型中的自噬、酸性细胞器和突触循环均有类似程度的减少。然后，我们开发了一种高通量筛选方法，并用慢病毒介导的CLN5或CLN6基因转移以及贝特类药物（吉非罗齐和非诺贝特）对CLN6缺陷的神经培养物中缺陷细胞的校正进行了测试。这些试验提供了一个简单的系统，可在体内测试之前快速筛选候选疗法或药物库。
