Design, Synthesis and Efficacy of New Small Molecule Therapeutics to Impede Myotonic Dystrophy

预防强直性肌营养不良的新型小分子疗法的设计、合成和功效

• 批准号: 10841887
• 负责人: Andrew Berglund
• 金额: $ 4.58万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10841887
• 关键词: 3' Untranslated Regions; Adult; Alternative Splicing; Biochemical; Biological Assay; Biology; Cell Culture Techniques; Cell Line; Cell physiology; Complex; Data; Disease; Engineering; Evaluation; Family; Generations; Genes; Genetic Transcription; Grant; In Vitro; Molecular; Muscular Dystrophies; Myotonic Dystrophy; Myotonic dystrophy type 1; Patients; Play; Polycyclic Compounds; Postbaccalaureate; Protein Family; Protein Kinase; Proteins; RNA; RNA Splicing; RNA-Binding Proteins; Regulation; Research Project Grants; Role; Series; Specificity; Structure; Systemic disease; Therapeutic; career development; design; effective therapy; genome-wide; improved; in vivo; member; research and development; skill acquisition; small molecule; small molecule therapeutics; therapeutic protein; tool; transcriptomics; treatment strategy

Project Abstract Myotonic dystrophy type 1 (DM1) is the leading cause of adult-onset muscular dystrophy, yet no effective treatment strategies are available to DM1 patients. The parental grant to this supplement project is aimed at understanding the disease mechanisms of myotonic dystrophy type 1 (DM1) and identifying potential therapeutics for this multisystemic disease. DM1 is caused by a CTG expansion repeat in the 3’ untranslated region (UTR) of the dystrophia myotonica protein kinase (DMPK) gene. When transcribed, the expanded repeats sequester muscleblind-like (MBNL) proteins, a family of alternative splicing regulators, disrupting their normal cellular splicing function. Our group has previously identified existing small molecules that rescue the molecular hallmarks of DM1, such as correction of aberrant splicing and reduction of ribonuclear foci and have now designed a series of new modified polycyclic compounds (MPCs) for evaluation. While we are currently investigating the therapeutic potential and mechanism of action of these new MPCs, preliminary data suggests that they operate through either reduction of the transcription of the toxic RNA and/or increases the expression of MBNL proteins. The project for this supplement is focused on complementary studies that seek to better understand the interaction between MBNL proteins, CUG expansion RNAs, and genome-wide splicing targets through the generation, analysis, and evaluation of various synthetic MBNL proteins. The Berglund lab has previous generated several synthetic MBNL proteins and demonstrated that these proteins show improved or altered splicing activity. These data suggested that synthetic MBNL may have the potential to be used as tools for investigating splicing regulation as well as for protein therapeutics for DM. Brianna’s project will extend and leverage these studies by generating additional synthetic MBNL proteins and probing their structure and function through in vitro biochemical assays and their impact on the RNA GOF in DM1 through transcriptomic studies in cell culture. This one-year post-baccalaureate project will provide research and career development skills around a central research project aim at: (1) identifying and characterizing in vitro the target engagement specificity of engineered synthetic MBNL in the context of myotonic dystrophy type 1 expansion RNAs; and (2) determining the in vivo transcriptomic specificity and therapeutic potential of engineered synthetic MBNL in DM1 patient- derived cell lines. Determining the functional and global transcriptomic impacts of synthetic MBNL proteins will enhance our understanding of the disease biology of DM while provide critical information to improve their therapeutic potential for the treatment of DM1 and potentially other splicing-related repeat expansion diseases.

项目摘要 肌发育症1型（DM1）是成​​年肌肉营养不良的主要原因，但无效 DM1患者可以使用治疗策略。该补充项目的父母赠款针对 了解1型肌发育症（DM1）的疾病机制并识别潜在的治疗 对于这种多系统疾病。 DM1是由3'未翻译区域（UTR）重复的CTG扩展引起的 营养不良的肌动蛋白蛋白激酶（DMPK）基因。转录时，扩展的重复测序仪 肌肉闪光（MBNL）蛋白质，一个替代剪接调节剂的家族，破坏了其正常的细胞 剪接功能。我们的小组以前已经确定了拯救分子的现有小分子 DM1的标志，例如校正异常剪接和核糖核心焦点的还原 设计了一系列新的修饰多环化合物（MPC）进行评估。当我们目前 初步数据研究了这些新MPC的治疗潜力和作用机理，表明 它们通过还原有毒RNA的转录和/或增加表达来运行 MBNL蛋白。该补充项目的项目集中于完整的研究，以寻求更好 了解MBNL蛋白，CUG膨胀RNA和全基因组剪接靶标之间的相互作用 通过各种合成MBNL蛋白的生成，分析和评估。 Berglund实验室有 以前产生了几种合成MBNL蛋白，并证明这些蛋白显示出改善或 改变剪接活动。这些数据表明合成MBNL可能有可能用作工具 研究剪接调节以及DM的蛋白质治疗。 Brianna的项目将扩展， 通过产生其他合成MBNL蛋白并探测其结构和功能来利用这些研究 通过体外生化测定及其对DM1中RNA GOF的影响 细胞培养。这项为期一年的学后项目将提供研究和职业发展技能 中央研究项目的目的是：（1）在体外识别和表征目标参与特异性的特殊性 在Myotonic营养不良1型膨胀RNA的背景下，设计了合成的MBNL； （2）确定 DM1患者中工程合成MBNL的体内转录组特异性和治疗潜力 派生的细胞系。确定合成MBNL蛋白的功能和全局转录组影响将 增强我们对DM疾病生物学的理解，同时提供关键信息以改善其 治疗DM1以及可能与剪接相关的重复扩张疾病的治疗潜力。