SMN associated proteins and compounds for SMA therapy

用于 SMA 治疗的 SMN 相关蛋白和化合物

• 批准号: 6540457
• 负责人: JIANHUA ZHOU
• 金额: $ 23.26万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6540457
• 关键词: DNA binding protein; RNA splicing; beta lactamase; biological signal transduction; degenerative motor system disease; disease /disorder model; gene deletion mutation; gene expression; genetically modified animals; green fluorescent proteins; immunoprecipitation; laboratory mouse; laboratory rabbit; messenger RNA; microarray technology; motor neurons; neuropharmacology; pathologic process; phosphatase inhibitor; progressive spinal muscular atrophy; protein structure function; small nuclear ribonucleoproteins; telomere; tissue /cell culture; yeast two hybrid system

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

描述（由申请人提供）： 常染色体隐性遗传脊髓性肌萎缩症（SMA）是最常见的疾病之一 婴儿死亡的遗传原因。 SMA 中存在前角细胞死亡和 肌肉无力。运动神经元存活基因 SMN 缺失或突变 都对疾病负有责任。有两个SMN基因。然而，仅 端粒拷贝（SMNt 或 SMNI）会导致疾病。由于单个核苷酸 差异，第二个基因SMN2中的T与SMNI中的C，大多数SMN2 mRNA 或蛋白质跳过外显子 7，导致 SMNA7 蛋白质不稳定并减少 其低聚能力。因此，SMA 中存在 SMN2 基因 患者无法弥补 SMNI 基因的缺失。要了解 SMA 的发病机制，该提案的首要目标是使用酵母 两种杂交筛选以鉴定 SMN 相互作用蛋白，特别是那些 来自运动神经元。相互作用将进一步通过其他特征来表征 互补方法，包括哺乳动物两种杂交测定、体外结合 测定和体内免疫共沉淀测定。生物学意义 SMN 及其相互作用因子之间的相互作用将在细胞中进行研究 线，并作为动物模型的长期目标。本次的第二个目标 提案是开发基于细胞的系统，用于基于 SMA 的治疗研究 假设来自 SMN2 的总或全长 SMN 蛋白增加 会降低 SMA 的严重程度。稳定细胞系和转基因小鼠 使用 GFP、荧光素酶或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）将用于鉴定小分子以促进外显子 7 包含在 SMN2 mRNA 和蛋白质。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 ZNS1 SRB R(01) 3 1 R01 NS41665-01 2000 年 12 月 13-14 日 周博士建华