Fluorescent IRE Sensor for Synucleinopathy Drug Discovery

用于突触核蛋白病药物发现的荧光 IRE 传感器

• 批准号: 10608833
• 负责人: Wen Shen
• 金额: $ 85.38万
• 依托单位: LUCERNA, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-22 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10608833
• 关键词: 5' Untranslated Regions; Affinity; Age; Alzheimer' s Disease; Amygdaloid structure; Amyloid beta-Protein Precursor; Antisense Oligonucleotides; Binding; Biological; Biological Assay; Buffers; Cells; Chemicals; Custom; Data; Dementia; Dementia with Lewy Bodies; Development; Disease; Disease Progression; Diversity Library; Elderly; Elements; Ensure; Fee-for-Service Plans; Future; Gene Duplication; Goals; Guidelines; In Vitro; Iron; Lewy Bodies; Lewy body pathology; Libraries; Life; Luciferases; Measurement; Messenger RNA; Multiple System Atrophy; National Center for Advancing Translational Sciences; Neurodegenerative Disorders; Neuroglia; Neurons; Onset of illness; Parkinson Disease; Parkinsonian Disorders; Pathogenicity; Pathologic; Performance; Phase; Population; Procedures; Production; Program Development; Protocols documentation; RNA; RNA-targeting therapy; Reagent; Reproducibility; Response Elements; Robotics; SNCA gene; Services; Severity of illness; Signal Transduction; Source; Specificity; Standardization; Structure; Symptoms; Testing; Therapeutic; Time; Translations; Validation; alpha synuclein; assay development; base; commercialization; dosage; drug discovery; experimental study; high throughput screening; improved; inhibitor; lead optimization; neuroprotection; prevent; programs; protein aggregation; sensor; small molecule; small molecule libraries; stem; success; synucleinopathy

ABSTRACT The goal of this Phase II proposal is to advance synucleinopathy disease drug discovery by validating a high- throughput screening (HTS)-ready assay and establish a RNA structure sensor platform for RNA-targeted drug discovery. Dementia with Lewy bodies is the second most common form of degenerative dementia in the elderly population after Alzheimer’s disease; and it is characterized by abnormal accumulation of alpha-synuclein (SNCA) aggregates. Diseases featuring pathogenic SNCA proteins are collectively known as synucleinopathies, which also include Parkinson’s disease, multiple system atrophy, and Alzheimer’s Disease with Amygdala restricted Lewy bodies. There is currently no disease-modifying cure available for any of the synucleinopathies. It is known that SNCA gene duplication increases SNCA levels and is correlated with disease progression and severity, leading to early parkinsonism and dementia. Studies showed reductions in SNCA levels can reduce aggregation, prevent Lewy body formation, and confer neuroprotection. Thus, inhibiting SNCA expression during disease prodromal phase has the potential to slow disease progression or halt disease onset. SNCA translation is controlled by an iron-response element (IRE) in the 5’UTR of the mRNA. To demonstrate feasibility, we developed proof-of-concept RNA structure sensors that were responsive to the binding of small molecules and antisense oligonucleotides, and demonstrated feasibility for HTS use. To accomplish the goal of this proposal, we will complete the following specific aims: 1) Finalize HTS optimization of the SNCA-specific RNA sensor and perform a pilot screen, 2) Establish the generalizability of the RNA structure sensor platform by developing HTS- compatible sensors targeting another pathogenic RNA structure, 3) Develop a standard operating procedure for the commercialization of custom RNA sensor services, 4) Perform a primary screen to identify inhibitors of SNCA protein translation. If successful, we will have a validated HTS assay for synucleinopathy drug discovery and a RNA structure sensor platform that aimed to accelerate the current pace in RNA structure-based drug discovery and to enable more RNA-targeted drug development programs targeting disease-causing RNA structures.

抽象的 该 II 期提案的目标是通过验证高通量来推进突触核蛋白病疾病药物的发现。 通量筛选 (HTS) 就绪分析并建立 RNA 靶向药物的 RNA 结构传感器平台 发现路易体痴呆是老年人退行性痴呆的第二常见形式。 阿尔茨海默病后的人群；其特征是α-突触核蛋白（SNCA）异常积累 以致病性 SNCA 蛋白为特征的疾病统称为突触核蛋白病。 还包括帕金森病、多系统萎缩症和杏仁核受限的阿尔茨海默病 目前已知对于任何突触核蛋白病都没有缓解疾病的方法。 SNCA 基因重复会增加 SNCA 水平并与疾病进展和严重程度相关， 研究表明，SNCA 水平的降低可以减少聚集， 防止路易体形成，并提供神经保护，从而在疾病期间抑制 SNCA 表达。 前驱期有可能减缓疾病进展或阻止疾病发作。 由 mRNA 5’UTR 中的铁反应元件 (IRE) 控制。 开发了概念验证 RNA 结构传感器，该传感器对小分子的结合做出响应 反义寡核苷酸，并证明了 HTS 使用的可行性。 我们将完成以下具体目标：1）完成SNCA特异性RNA传感器的HTS优化； 进行试点筛选，2) 通过开发 HTS- 建立 RNA 结构传感器平台的通用性 针对另一种致病性 RNA 结构的兼容传感器，3) 制定标准操作程序 定制 RNA 传感器服务的商业化，4) 进行初步筛选以识别 SNCA 抑制剂 如果成功，我们将拥有用于突触核蛋白病药物发现的经过验证的 HTS 测定和 RNA结构传感器平台旨在加快目前基于RNA结构的药物发现的步伐 并启用更多针对引起疾病的 RNA 结构的 RNA 靶向药物开发计划。