An analysis of molecular mechanism underlying spastin-induced spastic paraplegia and a strategy for the development of targeted therapies for the disease

spastin诱发痉挛性截瘫的分子机制分析及靶向治疗策略

基本信息

批准号：
18590954
负责人：
TAKIYAMA Yoshihisa
金额：
$ 2.48万
依托单位：
Jichi Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590954/
关键词：
Hereditary Spastic Paraplegia Spastin anti-spastin antibody siRNA Vinblastine SPG4 SPG7 Motor protein

项目摘要

The most common form of autosomal dominant hereditary spastic paraplegias (HSPs) is caused by mutations in the SPG4/SPAST gene, encoding spastin, a member of the AAA family of ATPases. Although spastin are suggested to be involved in microtubule dynamics, we have no due of the mechanism by which spastin mutations may lead to degeneration of corticospinal axons. Therefore, we investigated the function of spastin and the molecular mechanism underlying neurodegeneration due to spastin mutations. In addition, we attempted to find treatment agents for SPG4.(1) Subcellular localization of endogenous spastin :We investigated the subcellular localization of endogenous spastin in HeLa and NT2 cells using a spastin-specific antibody. In Hela cells, Spastin is predominantly localized in the nucleus during the interphase, with enrichment toward the centrosome and the spindle in the metaphase. In the telophase, spastin is concentrated in the nucleus and midzone region, with intense staining in the … More midbody. The results indicate that spastin plays an important role in cell division with microtubule severing process. In NT2 cells, spastin is enriched in the growth cone and in the branching regions. siRNA knock-down of spastin expression showed abnormal elongation and swelling of neurite, suggesting that spawn is essential far axon outgrouth.(2) Screening of possible treatment agents for SPG4We established a screening system (siRNA knock-down of spastin expression in NT2 and SHSY5Y cells) of possible treatment agents for SPG4. Then screening of possible treatment agents for SPG4 was performed using the system. Although vinblastin rescued the abnormal expansion of neurite, it also induced neuronal cell death.(3) Gene expression profiling on sacsinWe performed gene-profiling experiments to identify genes that are expressed at low levels together with siRNA knock-down of spastin expression. We found some candidate genes that could be associated with spastin. Among them, a gene was associated with the elongation of neurite and stability of microtubules, and was co-localized with spastin at the top of the neurite. To elucidate the net-work system associated with spastin will shed light on the establishment of treatment for SPG4. Less

常染色体显性遗传痉挛性截瘫（HSP）的最常见形式是由SPG4/SPAST基因突变引起的，该突变编码AAA AAA ATPase家族的spastin。尽管建议Spastin参与微管动力学，但我们没有造成痉挛突变可能导致皮质脊髓轴突变性的机制。因此，我们研究了Spastin的功能以及由于氨纶突变引起的神经退行性的分子机制。此外，我们试图找到用于SPG4的治疗剂。（1）内源性痉挛的亚细胞定位：我们使用翼盆特异性抗体研究了内源性痉挛在HELA和NT2细胞中内源性痉挛的亚细胞定位。在HeLa细胞中，Spastin在相间期间主要位于核中，并富集中心体和中期的纺锤体。在末期中，翼丁蛋白集中在细胞核和中区域，在…中部更强烈的染色。结果表明，用氨翼蛋白酶在微管几个过程中在细胞分裂中起重要作用。在NT2细胞中，翼丁蛋白富含生长锥和分支区域。 Spastin表达的siRNA敲除了神经蛋白的异常伸长和肿胀，这表明Spawn是必不可少的远轴突。然后使用系统对SPG4进行筛查。尽管vinblastin挽救了神经蛋白的异常膨胀，但它也诱导了神经元细胞死亡。（3）sacsinwe上的基因表达分析进行了基因促进实验，以鉴定与siRNA敲除spastin表达的siRNA敲低水平表达的基因。我们发现一些可能与Spastin相关的候选基因。其中，一个基因与神经蛋白的伸长和微管的稳定性有关，并与神经蛋白的顶部共定位。为了阐明与Spastin相关的网络系统，将阐明建立SPG4的治疗方法。较少的

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Autosomal recessive spastic ataxia of Chairlevoix-Saguenay(ARSACS)

常染色体隐性遗传的Chairlevoix-Saguenay 痉挛性共济失调（ARSACS）

DOI：
发表时间：
2006
期刊：
Neuropathology 26
影响因子：
0
作者：
Ito M;Suzuki Y;Okada T;Fukudome T;Masuda A;Yoshimura T;Takeda S;Krejci E;Ohno K;Takiyama Y
通讯作者：
Takiyama Y

McLeod syndrome mimicking muscular dystrophy

麦克劳德综合征模仿肌营养不良症