Harnessing the splicing code for targeted control of gene expression (UNLEASH)

mRNA前体的选择性剪接（AS）可以从根本上改变蛋白质表达、细胞表型和生理反应，在组织特异性基因调控和生物调控机制中起着重要作用。剪接的改变也有助于疾病机制，从神经变性到癌症。调节AS的药物最近为脊髓性肌萎缩症（一种常见的遗传性疾病）提供了第一种治疗方法，说明了治疗许多其他未满足需求的疾病的巨大潜力，只要我们了解控制剪接位点选择的机制以及如何用小分子调节它们。不幸的是，尽管经过几十年的研究，对控制AS特异性的机制缺乏全面的了解。这种基础知识上的差距阻碍了利用剪接调节剂作为研究基因功能、新疗法或其他生物技术应用的工具的机会。该项目直接解决了限制AS领域发展的主要技术挑战。基于广泛的初步数据，我们将使用多学科方法，结合化学、结构、细胞、系统生物学和机器学习来表征剪接位点选择的机制，并确定使用工具化合物调节这些机制的目标。这些结果将定义关键的调控序列、剪接因子和所涉及的分子相互作用，从而阐明剪接机制如何有效地适应，同时也区分广泛的剪接位点序列。这将使未来的应用程序能够利用剪接位点选择。我们的主要目标是回答核心问题，“是否通常有可能使用小分子以高特异性调节剪接？”成功将改变我们对人类基因表达的基本理解，并为制药公司开发新疗法带来重大机遇。

Alternative splicing (AS) of mRNA precursors plays important roles in tissue-specific gene regulation and biological regulatorymechanisms, as it can radically alter protein expression, cell phenotypes and physiological responses. Altered splicing also contributesto disease mechanisms, ranging from neurodegeneration to cancer. Drugs modulating AS have recently provided the first therapy forSpinal Muscular Atrophy, a common genetic disorder, illustrating the huge potential for treating many other diseases of unmet need,if only we understood the mechanisms controlling splice site selection and how to regulate them with small molecules.Unfortunately, despite decades of research, a comprehensive understanding of the mechanisms that control specificity of AS islacking. This gap in basic knowledge prevents opportunities to harness splicing modulators as tools to study gene function, noveltherapeutics or other biotech applications. This Project addresses head-on the major technical challenges that have limited progressin the AS field. Building on extensive preliminary data, we will use a multidisciplinary approach that combines chemical, structural,cellular, systems biology and machine learning to characterize mechanisms of splice site selection and identify targets for modulatingthese mechanisms using tool compounds. The outcomes will define key regulatory sequences, splicing factors and molecularinteractions involved, thereby illuminating how the splicing machinery efficiently accommodates, yet also discriminates between, awide range of splice site sequences. This will enable future applications harnessing splice site selection. Our primary goal is to answerthe central question, 'Is it generally possible to modulate splicing with high specificity using small molecules?' Success will transformour basic understanding of human gene expression and unleash major opportunities for Pharma to develop new therapeutics.