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 该第二阶段提案的目的是通过验证高 - 吞吐量筛选（HTS） - 就绪测定并建立靶向RNA靶向药物的RNA结构传感器平台 发现。痴呆症患有路易的身体是较早的第二常见退化性痴呆形式 阿尔茨海默氏病后的人口；它的特征是α-突触核蛋白（SNCA）异常积累 聚合。带有致病性SNCA蛋白的疾病统称为突触核酸，该疾病，该疾病 还包括帕金森氏病，多系统萎缩和杏仁核的阿尔茨海默氏病 路易的身体。目前尚无任何疾病改良治疗的任何突触核酸治疗。我们都知道 SNCA基因的重复增加了SNCA水平，并且与疾病的进展和严重程度相关， 导致早期帕金森氏症和痴呆症。研究表明，SNCA水平的降低可以减少聚集， 防止路易的身体形成和会议神经保护。那抑制疾病中SNCA的表达 前代具有减慢疾病进展或停止疾病发作的潜力。 SNCA翻译是 由mRNA 5'UTR中的铁反应元件（IRE）控制。为了证明可行性，我们 开发的概念验证RNA结构传感器对小分子的结合有响应 反义寡核苷酸，并证明了HTS使用的可行性。为了实现该提议的目标， 我们将完成以下特定目的：1）最终确定SNCA特异性RNA传感器的HTS优化和 执行试点屏幕，2）通过开发HTS-建立RNA结构传感器平台的普遍性 针对另一种致病RNA结构的兼容传感器，3） 定制RNA传感器服务的商业化，4）执行主要屏幕以识别SNCA的抑制剂 蛋白质翻译。如果成功的话，我们将有经过验证的HTS分析，用于发现突触核酸药物和A RNA结构传感器平台，旨在加速基于RNA结构的药物发现中的当前空间 并启用针对引起疾病的RNA结构的更多靶向RNA的药物开发计划。