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.

摘要 这项II期提案的目标是通过验证一种高水平的药物来推进突触核蛋白病药物的发现， 建立了一种用于RNA靶向药物的RNA结构传感器平台 的发现路易体痴呆是老年人中第二种最常见的退行性痴呆 阿尔茨海默病后的人群;其特征是α-突触核蛋白（SNCA）的异常积累 集料.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、多系统萎缩和杏仁核受限的阿尔茨海默病 路易体目前还没有针对任何突触核蛋白病的疾病修饰疗法。已知 SNCA基因重复会增加SNCA水平，并与疾病进展和严重程度相关， 导致早期帕金森症和痴呆症研究表明，SNCA水平的降低可以减少聚集， 防止路易体形成，并赋予神经保护作用。因此，在疾病期间抑制SNCA表达， 前驱期具有减缓疾病进展或停止疾病发作的潜力。SNCA翻译 由mRNA的5 'UTR中的铁反应元件（IRE）控制。为了证明可行性，我们 开发了概念验证的RNA结构传感器，该传感器对小分子的结合有反应， 反义寡核苷酸，并证明了HTS使用的可行性。为了实现本提案的目标， 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， 进行中试筛选，2）通过开发HTS- 针对另一种病原性RNA结构的兼容传感器，3）开发标准操作程序， 定制RNA传感器服务的商业化，4）进行初步筛选以鉴定SNCA的抑制剂 蛋白质翻译如果成功，我们将有一个有效的HTS检测突触核蛋白病药物发现和 RNA结构传感器平台，旨在加快基于RNA结构的药物发现的当前步伐 并使更多的RNA靶向药物开发计划靶向致病RNA结构。