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，存在前角细胞死亡， 肌肉无力运动神经元存活基因SMN的缺失或突变， 是导致这种疾病的原因有两个SMN基因。但只有 端粒复制（SMNt或SMNI）导致疾病。由于单个核苷酸 差异，T在第二基因SMN 2中，C在SMNI中，大部分SMN 2 mRNA 或蛋白质跳过外显子7，导致不稳定的SMNA 7蛋白和减少 它的低聚能力。因此，SMA中SMN 2基因的存在 患者不能弥补SMNI基因的缺失。了解 SMA的发病机制，这项建议的第一个目标是使用酵母 双杂交筛选以鉴定SMN相互作用蛋白，特别是那些 从运动神经元。这些相互作用将进一步由其他 互补方法，包括哺乳动物双杂交测定、体外结合 测定和体内免疫共沉淀测定。的生物学意义 SMN和它的相互作用者之间的相互作用将在细胞中被研究 线，并作为长期目标，在动物模型。第二个目标是 建议是开发基于细胞的系统，用于基于SMA的治疗研究。 假设从SMN 2中增加总的或全长SMN蛋白 可以降低SMA的严重程度。稳定的细胞系和转基因小鼠 表达具有报告基因如GFP、荧光素酶或 将建立β-内酰胺酶。高通量筛选和低通量筛选（HTS， LTS）将用于鉴定小分子以促进外显子7的包含， SMN 2 mRNA和蛋白。这些化合物将在SMA小鼠模型中进行测试。 调节SMN基因RNA剪接的信号通路和其他机制 将被调查。 1 ZNS 1 SRB R（01） 3 1 R01 NS41665-01 二○年十二月十三日至十四日 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