Signature-based chemical screening for DYT6 dystonia

基于特征的 DYT6 肌张力障碍化学筛查

• 批准号: 8112220
• 负责人: David Cristopher Bragg
• 金额: $ 26.55万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8112220
• 关键词: Acute Myelocytic Leukemia; Affect; Apoptosis; Biological Assay; Biological Models; Cells; Cephalic; Chemicals; Classification; Collection; DNA Binding; DNA Microarray Chip; DNA-Binding Proteins; DYT6 gene; Defect; Disease; Dystonia; Ewings sarcoma; Future; Gene Expression; Gene Expression Profile; Generic Drugs; Genes; Genotype; Hereditary Dystonia; Individual; Inherited; Larynx; Link; Malignant Neoplasms; Methods; Metric; Microspheres; Movement Disorders; Muscle Contraction; Mutation; Outcome; Pathogenesis; Patients; Performance; Phenotype; Population Heterogeneity; Proteins; Relative (related person); Reproducibility; Screening procedure; Smooth Muscle; Speech; Sum; System; Technology; Tertiary Protein Structure; Therapeutic; Training; Transcript; Upper Extremity; Variant; base; cost; drug candidate; drug discovery; genome-wide; lymphoblast; meetings; novel; small molecule; transcription factor

DESCRIPTION (provided by applicant): DYT6 dystonia is a hereditary movement disorder for which few treatment options exist. Affected individuals develop involuntary muscle contractions, primarily of upper limbs and cranial region with frequent speech defects due to laryngeal dystonia. The causative gene was recently identified as THAP1 which encodes a DNA binding protein, THAP1 (thanatos-associated protein [THAP] domain-containing apoptosis-associated protein- 1). Multiple groups have now independently linked 35 THAP1 mutations to dystonia in genetically diverse populations throughout the world, thereby revealing DYT6 as a substantial cause of familial dystonia. Most mutations impact residues known to be critical for THAP1's DNA binding activity, raising the hypothesis that DYT6 pathogenesis may involve aberrant transcriptional activity due to insufficient levels of functional THAP1 protein. Consistent with that hypothesis, we have detected a transcriptional phenotype in lymphoblasts bearing one of the DYT6 mutations, relative to control cells. In this project we propose to develop and pilot a novel assay for identifying new drug candidates for treating DYT6. The assay uses technology that has been largely applied to drug discovery in cancers similarly linked to aberrant transcription factor activity, such as acute myeloid leukemia (AML) and Ewing sarcoma. Using DNA microarrays, we will first define a gene expression signature distinguishing patient from control cells, using lines representing 18 DYT6 genotypes to find common markers representative of the DYT6 disease state. That signature will then be converted to a low cost assay based on the Luminex FlexMAP" system, which provides an automated method for capturing and quantifying target transcripts from cells in a conventional cell-based assay format. After validating that the Luminex assay recapitulates the microarray signature with sufficient reproducibility and Z' factor, we will pilot the screen in a mixed collection of 15,000 small molecules to seek compounds that normalize the DYT6 transcriptional phenotype. We expect potential outcomes of this project to be: (1) a validated assay that could be used for a large scale screening campaign to find novel DYT6 therapeutics (2) an established signature- based screening platform that could be generalized to other forms of hereditary dystonia; and (3) candidates from the pilot screen to be further characterized in DYT6 model systems.

描述（由申请人提供）：DYT6肌张力障碍是一种遗传运动障碍，几乎没有治疗选择。受影响的个体出现非自愿性肌肉收缩，主要是上肢和颅区域，由于喉肌肌张力障碍而经常出现语音缺陷。最近将致病基因鉴定为THAP1，它编码DNA结合蛋白THAP1（TASATOS相关蛋白[THAP]含有域的细胞凋亡相关蛋白-1）。现在，多个群体已独立地将35个THAP1突变与全世界种类多样的人群中的肌张力障碍联系起来，从而揭示了Dyt6是家族性肌张力障碍的实质性原因。大多数突变会影响已知的残基对THAP1的DNA结合活性至关重要，这提出了以下假设：Dyt6发病机理可能涉及由于功能性THAP1蛋白水平不足而导致的异常转录活性。与该假设一致，我们在淋巴细胞中检测到具有DYT6突变之一的淋巴细胞中的转录表型，相对于对照细胞。在这个项目中，我们建议开发和试行一种新的测定法，以鉴定用于治疗DYT6的新药物。该测定法使用与异常转录因子活性相似的癌症中的药物发现，例如急性髓性白血病（AML）和ewing肉瘤。使用DNA微阵列，我们首先使用代表18个DYT6基因型的线来定义将患者与对照细胞区分开的基因表达，以找到代表Dyt6疾病状态的公共标记。 That signature will then be converted to a low cost assay based on the Luminex FlexMAP" system, which provides an automated method for capturing and quantifying target transcripts from cells in a conventional cell-based assay format. After validating that the Luminex assay recapitulates the microarray signature with sufficient reproducibility and Z' factor, we will pilot the screen in a mixed collection of 15,000 small molecules to seek compounds that normalize the DYT6转录表型。