SMN associated proteins and compounds for SMA therapy

描述(由申请人提供)： 常染色体隐性遗传性脊髓性肌萎缩症(SMA)是最常见的 婴儿死亡的遗传原因。在SMA中，有前角细胞死亡和 肌肉无力。存活运动神经元基因SMN的缺失或突变， 是疾病的罪魁祸首。有两个SMN基因。然而，只有 端粒复制(SMNt或SMNI)导致疾病。由于单核苷酸 不同的是，在SMNI中，第二个基因SMN_2中的T与C中的SMN_2m RNA居多 或蛋白质跳过外显子7，导致不稳定的SMNA7蛋白质和减少 它的齐聚能力。因此，SMN2基因在SMA中的存在 患者无法补偿SMNI基因的丢失。要了解 SMA的发病机制，这个建议的第一个目标是使用酵母 双杂交筛选鉴定SMN相互作用蛋白，特别是那些 来自运动神经元。这些相互作用将进一步由其他 补充方法，包括哺乳动物双杂交分析，体外结合 免疫共沉淀法和体内共沉淀法。它的生物学意义 将在细胞中研究SMN与其相互作用因子之间的相互作用 在动物模型中，作为长期目标。这方面的第二个目标 建议为以SMA为基础的治疗研究开发基于细胞的系统 关于SMN的全长或全长SMN蛋白增加的假设 会减轻SMA的严重程度。稳定的细胞系和转基因小鼠 用GFP、荧光素酶或其他报告基因表达外显子7剪接盒 将建立β-内酰胺酶。高通量和低通量筛选(HTS， LTS)将被用来鉴定小分子以促进外显子7在 SMN2mRNA和蛋白的表达。这些化合物将在SMA小鼠模型中进行测试。 调控SMN基因RNA剪接的信号通路和其他机制 将会被调查。 1 ZNS1 SRB R(01)3 1 R01 NS41665-01 2000年12月13日至14日，周建华博士

DESCRIPTION (provided by applicant): The autosomal recessive spinal muscular atrophy (SMA) is one of the most common genetic causes of infant death. In SMA, there is anterior horn cell death and muscle weakness. Deletions or mutations in the survival motor neuron gene, SMN, are responsible for the disease. There are two SMN genes. However, only telomeric copy (SMNt or SMNI) causes disease. Due to a single nucleotide difference, T in the second gene SMN2 from C in SMNI, the majority of SMN2 mRNA or protein skips exon7, resulting in an unstable SMNA7 protein and reduction of its oligomerization ability. Therefore, the presence of the SMN2 gene in SMA patients can not compensate for the loss of the SMNI gene. To understand the pathogenesis of SMA, the first goal of this proposal is to use the yeast two-hybrid screens to identify SMN interacting proteins, particularly those from motor neurons. The interactions will be further characterized by other complementary methods including mammalian two hybrid assays, in vitro binding assays and in vivo co-immunoprecipitation assays. The biological significance of interactions between SMN and its interactors will be investigated in cell lines, and as long-term goals, in animal models. The second goal of this proposal is to develop cell-based systems for therapeutic studies of SMA based on the hypothesis that increasing of total or full-length SMN protein from SMN2 would reduce the severity of SMA. Stable cell lines and transgenic mice expressing exon 7 splicing cassettes with reporters such as GFP, luciferase or P-lactamase will be established. Both high and low throughput screening (HTS, LTS) will be used to identify small molecules to promote inclusion of exon 7 in SMN2 mRNA and protein. These compounds will be tested in SMA mouse models. Signal pathways and other mechanisms that regulate RNA splicing of SMN genes will be investigated. 1 ZNS1 SRB R(01) 3 1 R01 NS41665-01 DECEMBER 13-14, 2000 ZHOU, DR. JIANHUA