Discovery of Small Molecules that Block Supt4h1-Supt5h Dimerization for Potential C9FTD/ALS Therapeutics

发现阻止 Supt4h1-Supt5h 二聚化的小分子，用于潜在的 C9FTD/ALS 治疗

基本信息

批准号：
9809219
负责人：
Keith Thomas Gagnon
金额：
$ 8.79万
依托单位：
SOUTHERN ILLINOIS UNIVERSITY CARBONDALE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9809219
关键词：
Adult Affect Affinity Animal Model Base Sequence Binding Biological Biological Assay Biological Sciences Bypass C9ORF72 Cells Chemical Structure Collaborations Complex Crystallization Defect Development Dimerization Disease Disease Marker Disease Progression Docking Drug Delivery Systems Drug Kinetics Drug Screening Enzymes Event Future Gene Expression Genes Genetic Genetic Transcription Genome Goals Huntington Disease In Vitro Inherited Lactamase Lactams Lead Libraries Measures Mediating Modeling Molecular Molecular Disease Mus Mutagenesis Mutation N-terminal Nature Neurologic Nucleic Acids Orthologous Gene Pathology Patients Pharmaceutical Preparations Pharmacodynamics Production Property Proteins RNA RNA Polymerase II Repetitive Sequence Reporter Reporting Route Specificity Structural Protein Structure Structure-Activity Relationship Tandem Repeat Sequences Testing Therapeutic Titrations Toxic effect Toxicity Tests Translations Treatment Efficacy Universities Yeasts base beta-Lactamase c9FTD/ALS cost effective curative treatments design dimer drug candidate drug development effective therapy fly frontotemporal lobar dementia-amyotrophic lateral sclerosis gain of function high throughput screening improved inhibitor/antagonist innovation knock-down molecular marker mouse model mutant nervous system disorder protein structure reconstitution screening simulation small molecule small molecule libraries transcription factor

项目摘要

PROJECT SUMMARY C9FTD/ALS is a rapidly progressive and debilitating neurological disease caused by expansion of a simple tandem repeat sequence in the gene C9ORF72. C9FTD/ALS is the number one inherited cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). There are no effective treatments for C9FTD/ALS, nor the other neurological repeat expansion disorders In C9FTD/ALS and most other repeat expansion disorders, of which over two dozen exist, the repeat expansion is transcribed into expanded tandem repeat-containing RNA, or xtrRNA, which mediates the molecular mechanisms of disease. Although the molecular chain of events that lead to C9FTD/ALS pathology are still unclear, it is widely accepted that reducing or blocking production of the xtrRNA will lead to effective therapeutic treatments that can halt disease progression. The key will lie in identifying treatments that can selectively inhibit transcription of large repeat expansions without affecting normal gene expression. A protein called Supt4h1 (Spt4 in yeast) acts as a processivity factor to the core RNA polymerase II enzyme to improve transcription across repetitive, structurally complex, or large regions of the genome. Previous studies have demonstrated that Supt4h1 is largely dispensable in yeast and its knock-down in C9FTD/ALS model organisms and patient-derived cells significantly mitigates disease pathology. Supt4h1 interacts with RNA polymerase II through dimerization with Sup5h, a core transcription factor. A crystal structure of this dimer reveals precise molecular contacts. Thus, we will develop an Supt4h1- Supt5h dimerization assay that can rapidly report dimerization status and is amenable to high throughput drug screening. The assay is colorimetric and quick, making high throughput chemical library screening accessible and cost-effective. We will search for small molecules that inhibit dimerization. After counter- screening and titration of the best inhibitors, we will further characterize the ability of top molecules to block C9ORF72 xtrRNA transcription and downstream molecular markers of disease pathology in patient- derived cells. This project will potentially deliver lead small molecules for further development as drug candidates for C9FTD/ALS. This project may also help fill an urgent gap in therapeutics for a number of diseases comprising an entire class of neurological disorders. The Supt4h1-Supt5h dimerization assay developed here will be useful for screening other libraries of small molecules or active biologics.

项目摘要 C9FTD/ALS是一种迅速进行性和使人衰弱的神经系统疾病。基因C9orf72中的简单串联重复序列。 c9ftd/als是第一的继承原因额颞痴呆（FTD）和肌萎缩性侧索硬化症（ALS）。没有有效的治疗对于C9FTD/ALS或其他神经系统重复扩张障碍在C9FTD/ALS和大多数其他重复膨胀障碍中，其中有二十多个，重复膨胀被转录为扩展的串联重复的RNA或XTRRNA，该RNA介导疾病的分子机制。虽然导致C9FTD/ALS的事件的分子链病理尚不清楚，已广泛认为，减少或阻止XTRRNA的产生将导致有效的治疗性治疗可以阻止疾病进展。关键在于识别可以选择性地抑制大重复扩张的转录而不会影响正常的治疗方法基因表达。一种称为SUPT4H1（酵母中SPT4）的蛋白质是核心RNA聚合酶II的加工性因子酶改善基因组的重复，结构复杂或大区域的转录。先前的研究表明，Supt4H1在酵母中基本上是可赋入的，并且其倒下 C9FTD/ALS模拟生物和患者衍生细胞可显着减轻疾病病理。 SUPT4H1通过与核心转录因子SUP5H二聚化与RNA聚合酶II相互作用。该二聚体的晶体结构揭示了精确的分子接触。因此，我们将开发一个Supt4H1- SUPT5H二聚化测定可以快速报告二聚化状态，并且可以适应高吞吐量药物筛查。该测定是比色的，快速的，可以使高吞吐量化学库筛选可访问且具有成本效益。我们将搜索抑制二聚化的小分子。反击之后 - 筛选和滴定最佳抑制剂，我们将进一步表征顶部分子的能力 C9orf72 XTRRNA转录和疾病病理学病理学的下游分子标记衍生细胞。该项目将有可能作为候选药物的进一步发展提供铅小分子 C9FTD/ALS。该项目还可能有助于填补许多疾病的紧急治疗差异包括整个神经系统疾病。 SUPT4H1-SUPT5H二聚化测定开发了这里将有助于筛选其他小分子或活性生物制剂的文库。