Dissecting Signalling Pathways with PROTAC Chemical Probes.

使用 PROTAC 化学探针剖析信号通路。

• 批准号: 2441588
• 金额: --
• 依托单位: University of Southampton
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2441588
• 关键词: Dissecting; Signalling; Pathways; PROTAC; Chemical

Proteolysis targeting chimeras (PROTACs) are hetero-bifunctional molecules where one fragment interacts with the protein of interest (POI) and the other binds to a component of an ubiquitin ligase. This results in the poly-ubiquitination of the POI, which then undergoes irreversible proteasomal degradation, in a catalytic manner. This strategy to reduce cellular protein levels has generated huge interest and excitement in biology, and at present, there is significant research aimed at developing PROTACs to target proteins and signalling pathways previously thought undruggable using classical protein-protein interaction inhibitors. This project aims to develop/characterise new classes of cell permeable PROTACs, as novel probes to interrogate important signalling pathways and their key protein effectors. Examples include the Wnt/Beta-catenin signalling pathway and chromatin remodelling complexes, which play multiple pivotal functions in human physiology. These critical roles are underscored by the observation that their dysregulation leads to multiple cancer types. First, the candidate will focus on the development of PROTAC probes targeting key POIs at regulatory nodes. This will involve the molecular design, chemical synthesis and in vitro biophysical characterisation (NMR, ITC, MS) of the PROTACs. A second objective will focus on elucidating the cellular mode of action of the new PROTACs. Their selectivity and ability to deplete the POIs will be quantified by WB and live imaging using GFP constructs; and their downstream effects determined by monitoring mRNA levels of downstream genes known to respond to POI inhibition across a panel of cancer cell lines. This project builds on i)recent proof-of-concept results in the Baud and Linclau laboratories showing that it is possible to enhance PROTAC properties (e.g. solubility/lipophilicity) and selectivity through synthetic alteration, circumventing an important historical bottleneck in the field; and ii)expertise in cancer cell biology and signalling in the Giamas laboratory (e.g. identification/deciphering of new genes/proteins implicated in important biological processes).

蛋白水解靶向嵌合体(PROTAC)是一种异质双功能分子，其中一个片段与目的蛋白(POI)相互作用，另一个片段与泛素连接酶的一个组分结合。这导致POI的多泛素化，然后以催化的方式经历不可逆转的蛋白酶体降解。这种降低细胞蛋白质水平的策略在生物学领域引起了巨大的兴趣和兴奋，目前，有一些重要的研究旨在开发PROTAC来靶向蛋白质和信号通路，以前人们认为使用经典的蛋白质-蛋白质相互作用抑制剂是无法治愈的。该项目旨在开发/表征新型的细胞通透性PROTAC，作为询问重要信号通路及其关键蛋白效应器的新探针。例如Wnt/Beta-catenin信号通路和染色质重塑复合体，它们在人体生理学中发挥着多种关键功能。观察到它们的失调会导致多种癌症类型，这突显了这些关键作用。首先，候选人将专注于开发针对监管节点关键POI的PROTAC探测器。这将涉及到PROTAC的分子设计、化学合成和体外生物物理特性(核磁共振、ITC、MS)。第二个目标将集中在阐明新的PROTAC的细胞作用模式。它们消耗POI的选择性和能力将通过WB和使用GFP结构的实时成像来量化；它们的下游效应将通过监测一组癌细胞系中已知对POI抑制做出反应的下游基因的mRNA水平来确定。该项目的基础是：i)波德和林克劳实验室最近的概念验证结果表明，有可能通过合成改变提高PROTAC的性质(例如，溶解性/亲脂性)和选择性，从而绕过该领域的一个重要历史瓶颈；以及ii)Giamas实验室在癌细胞生物学和信号传递方面的专业知识(例如，识别/破译与重要生物过程有关的新基因/蛋白质)。