Low dose tubulin-binding drugs rescue peroxisome trafficking deficit in patient-derived stem cells in Hereditary Spastic Paraplegia.
Low dose tubulin-binding drugs rescue peroxisome trafficking deficit in patient-derived stem cells in Hereditary Spastic Paraplegia.
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DOI：
10.1242/bio.20147641
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发表时间：
2014-05-23
期刊：
影响因子：
2.4
通讯作者：
Mackay-Sim A
中科院分区：
文献类型：
--
作者：
Fan Y;Wali G;Sutharsan R;Bellette B;Crane DI;Sue CM;Mackay-Sim A
遗传性痉挛截瘫(HSP)是一组遗传异质性疾病，诊断为进行性步态障碍伴肌肉无力和痉挛，目前尚无针对潜在病理生理学的治疗方法。Spastin的突变是HSP的常见原因。Spastin是一种微管切断蛋白，其突变在小鼠体内会导致轴突运输缺陷。在人类患者来源的嗅神经球来源的细胞中，痉挛蛋白突变导致乙酰化α-微管蛋白水平较低，而乙酰化微管蛋白是稳定微管的标志，并导致过氧化物体运输速度放缓。在这里，我们筛选了四种不同浓度的微管蛋白结合药物，以观察它们在患者来源的神经节细胞中拯救乙酰化α-微管蛋白水平的能力。然后选择将乙酰化的α-微管蛋白恢复到对照来源的ONS细胞中水平的药物剂量，以评估它们拯救过氧化物酶运输缺陷的能力。自动显微镜筛选发现，四种药物(0.5 NM紫杉醇、0.5 NM长春花碱、2 NM Epothilone D、10 µM noskine)可以挽救患者来源的ONS细胞中的乙酰化α-微管蛋白，剂量非常低。这些相同的剂量挽救了过氧化物体运输缺陷，将过氧化物体的速度恢复到未经处理的对照细胞水平。这些结果证明了一种新的药物筛选方法，该方法基于高通量自动显微镜对乙酰化α-微管蛋白进行药物筛选，然后对基于微管的过氧化物体运输进行功能验证。从临床角度来看，所有测试的药物都用于临床，但剂量要高得多。重要的是，Epothilone D和noscape ine可以进入中枢神经系统，使它们成为未来临床试验的潜在候选者。
Hereditary Spastic Paraplegia (HSP) is a genetically heterogeneous group of disorders, diagnosed by progressive gait disturbances with muscle weakness and spasticity, for which there are no treatments targeted at the underlying pathophysiology. Mutations in spastin are a common cause of HSP. Spastin is a microtubule-severing protein whose mutation in mouse causes defective axonal transport. In human patient-derived olfactory neurosphere-derived (ONS) cells, spastin mutations lead to lower levels of acetylated α-tubulin, a marker of stabilised microtubules, and to slower speed of peroxisome trafficking. Here we screened multiple concentrations of four tubulin-binding drugs for their ability to rescue levels of acetylated α-tubulin in patient-derived ONS cells. Drug doses that restored acetylated α-tubulin to levels in control-derived ONS cells were then selected for their ability to rescue peroxisome trafficking deficits. Automated microscopic screening identified very low doses of the four drugs (0.5 nM taxol, 0.5 nM vinblastine, 2 nM epothilone D, 10 µM noscapine) that rescued acetylated α-tubulin in patient-derived ONS cells. These same doses rescued peroxisome trafficking deficits, restoring peroxisome speeds to untreated control cell levels. These results demonstrate a novel approach for drug screening based on high throughput automated microscopy for acetylated α-tubulin followed by functional validation of microtubule-based peroxisome transport. From a clinical perspective, all the drugs tested are used clinically, but at much higher doses. Importantly, epothilone D and noscapine can enter the central nervous system, making them potential candidates for future clinical trials.
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影响因子：
4.3
作者：
Abrahamsen G;Fan Y;Matigian N;Wali G;Bellette B;Sutharsan R;Raju J;Wood SA;Veivers D;Sue CM;Mackay-Sim A
通讯作者：
Mackay-Sim A
影响因子：
3
作者：
Aneja, Ritu;Dhiman, Neerupma;Joshi, Harish C.
通讯作者：
Joshi, Harish C.
影响因子：
5.2
作者：
Denton, Kyle R.;Lei, Ling;Grenier, Jeremy;Rodionov, Vladimir;Blackstone, Craig;Li, Xue-Jun
通讯作者：
Li, Xue-Jun
影响因子：
4
作者：
Nguyen, T;Bjorkman, J;Crane, DI
通讯作者：
Crane, DI
影响因子：
8.6
作者：
Duning, Thomas;Warnecke, Tobias;Deppe, Michael
通讯作者：
Deppe, Michael