National Centre for Adult Stem Cell Research, Eskitis Institute for Cell and Molecular Therapies, Griffith University, Brisbane, Queensland 4111, Australia.
Dis Model Mech. 2013 Mar;6(2):489-502. doi: 10.1242/dmm.010884. Epub 2012 Dec 20.
Hereditary spastic paraplegia (HSP) leads to progressive gait disturbances with lower limb muscle weakness and spasticity. Mutations in SPAST are a major cause of adult-onset, autosomal-dominant HSP. Spastin, the protein encoded by SPAST, is a microtubule-severing protein that is enriched in the distal axon of corticospinal motor neurons, which degenerate in HSP patients. Animal and cell models have identified functions of spastin and mutated spastin but these models lack the gene dosage, mutation variability and genetic background that characterize patients with the disease. In this study, this genetic variability is encompassed by comparing neural progenitor cells derived from biopsies of the olfactory mucosa from healthy controls with similar cells from HSP patients with SPAST mutations, in order to identify cell functions altered in HSP. Patient-derived cells were similar to control-derived cells in proliferation and multiple metabolic functions but had major dysregulation of gene expression, with 57% of all mRNA transcripts affected, including many associated with microtubule dynamics. Compared to control cells, patient-derived cells had 50% spastin, 50% acetylated α-tubulin and 150% stathmin, a microtubule-destabilizing enzyme. Patient-derived cells were smaller than control cells. They had altered intracellular distributions of peroxisomes and mitochondria and they had slower moving peroxisomes. These results suggest that patient-derived cells might compensate for reduced spastin, but their increased stathmin expression reduced stabilized microtubules and altered organelle trafficking. Sub-nanomolar concentrations of the microtubule-binding drugs, paclitaxel and vinblastine, increased acetylated α-tubulin levels in patient cells to control levels, indicating the utility of this cell model for screening other candidate compounds for drug therapies.
遗传性痉挛性截瘫(HSP)导致进行性步态障碍,下肢肌肉无力和痉挛。 SPAST 的突变是成人发病、常染色体显性遗传性 HSP 的主要原因。 Spastin 是由 SPAST 编码的蛋白质,是一种微管切割蛋白,在皮质脊髓运动神经元的远端轴突中丰富,在 HSP 患者中会退化。动物和细胞模型已经确定了 spastin 和突变 spastin 的功能,但这些模型缺乏疾病患者所具有的基因剂量、突变可变性和遗传背景。在这项研究中,通过比较来自健康对照者嗅粘膜活检的神经祖细胞与具有 SPAST 突变的 HSP 患者的类似细胞,涵盖了这种遗传变异性,以确定 HSP 中改变的细胞功能。与对照细胞相比,患者衍生的细胞在增殖和多种代谢功能方面与对照细胞相似,但基因表达存在主要失调,影响了 57%的所有 mRNA 转录物,包括许多与微管动力学相关的转录物。与对照细胞相比,患者衍生的细胞具有 50%的 spastin、50%的乙酰化α-微管蛋白和 150%的 stathmin,这是一种微管不稳定酶。患者衍生的细胞比对照细胞小。它们具有改变的过氧化物酶体和线粒体的细胞内分布,并且过氧化物酶体的运动速度较慢。这些结果表明,患者衍生的细胞可能会补偿减少的 spastin,但增加的 stathmin 表达减少了稳定的微管并改变了细胞器运输。亚纳摩尔浓度的微管结合药物紫杉醇和长春碱增加了患者细胞中乙酰化α-微管蛋白的水平,使其达到对照水平,表明该细胞模型可用于筛选其他候选化合物进行药物治疗。