Discovery of small molecule-mediated protein degradation pathways in human cancer

人类癌症中小分子介导的蛋白质降解途径的发现

基本信息

批准号：
10576976
负责人：
Xiaoyu Zhang
金额：
$ 24.9万
依托单位：
NORTHWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10576976
关键词：
Advisory Committees Antineoplastic Agents Binding Binding Proteins Biological Biological Process Biology CRISPR/Cas technology Cancer Cell Growth Cancer cell line Catalysis Cell Nucleus Cell physiology Cells Cellular biology ChIP-seq Chemicals Chromatin Clinical Trials Communication Cytosol Degradation Pathway Development Drug Design Educational process of instructing Educational workshop Environment Enzymes Gene Expression Genetic Transcription Glues Goals Grant Human Institution Knowledge Laboratories Ligands Luciferases MDA MB 231 Malignant Neoplasms Maps Marketing Mass Spectrum Analysis Mediating Membrane Mentors Methods Molecular Molecular Biology Nuclear Pharmaceutical Preparations Phase Phenotype Process Property Protein Degradation Induction Proteins Proteome Proteomics Receptor Signaling Research Research Institute Role Scientist Site-Directed Mutagenesis Substrate Specificity System Tacrolimus Binding Protein 1A Technology Training Training Programs Ubiquitin Ubiquitination cancer type career development cell growth chemoproteomics cytotoxicity drug action experience gain of function human disease multicatalytic endopeptidase complex new therapeutic target next generation sequencing pharmacologic protein degradation protein expression protein function screening skills small molecule small molecule inhibitor small molecule libraries stable cell line tool transcriptome sequencing ubiquitin-protein ligase

项目摘要

Ligand-dependent protein degradation has emerged as a compelling strategy to pharmacologically control the protein content of cells. This strategy has advantages that include converting silent ‘protein-binding’ small molecules to ‘protein degraders’, which has the potential to substantially increase the number of druggable proteins in human disease such as cancer, and to operate in a catalytic manner that may lower the drug concentrations required to produce a pharmacological effect. Nonetheless, until recently, only a handful of the 600+ human E3 ligases have been found to support this process. Importantly, these E3 ligases have been found to show distinct and restricted substrate specificities, underscoring the need to discover additional ligandable E3 ligases with differentiated properties to realize the full scope of targeted protein degradation as a pharmacological strategy. We recently leveraged chemoproteomic platforms and molecular biology approaches to identify DCAF16, a poorly characterized E3 ligase, as a target of electrophilic probes that promotes the nuclear-restricted degradation of proteins. Importantly, DCAF16 is capable of supporting ligand- induced protein degradation at low fractional engagement (10-40%), which may enable the degradation of target proteins while minimally perturbing its endogenous substrates. In this proposal, by integrating chemoproteomic platforms with cell biology and molecular biology approaches, I plan to 1) identify endogenous substrates and biological functions of DCAF16, 2) screen and identify small molecules that disrupt cancer cell growth by engaging DCAF16 and inducing neo-substrate degradation, and 3) discover additional E3 ligases with distinct and restricted expressions in different cancer types that support ligand-induced targeted protein degradation. My long-term goals are to develop potent and selective chemical probes for DCAF16 and other druggable E3 ligases and to develop pharmacological tools to intervene hard-to-drug or even ‘undruggable’ cancer targets. The proposed studies will be carried out at The Scripps Research Institute, a top-ranking research institution which combines cutting edge biological and chemical research to offer a unique cross- disciplinary scientific environment. To build a successful career development and training program, I will attend an array of TSRI- or UCSD-hosted courses, workshops and seminars that will comprehensively advance my skills in mentoring, teaching, scientific communication, critical assessment and laboratory management. I will benefit tremendously from the mentored phase training with Dr. Benjamin Cravatt who has over 20 years of experiences in developing small molecule inhibitors of protein targets as well as chemical proteomic technologies, Dr. Michael Erb who has expertise in transcriptional biology in cancer, and my advisory committee, which includes prominent scientists – Dr. Jeffery Kelly and Dr. Dale Boger. The proposed studies will not only push the boundaries of our knowledge of targeted protein degradation, but may also be the starting point for the development of novel therapeutics targeting important yet undruggable proteins in cancer.

配体依赖性蛋白质降解已成为一种令人信服的药理学控制策略该策略的优点包括将沉默的“蛋白质结合”转化为小分子。分子到“蛋白质降解剂”，这有可能大幅增加可成药的数量人类疾病（如癌症）中的蛋白质，并以催化方式发挥作用，可能会降低药物浓度然而，直到最近，产生药理作用所需的浓度也只有少数。已发现 600 多种人类 E3 连接酶支持此过程。重要的是，这些 E3 连接酶已被证实。发现显示出明显且受限的底物特异性，强调需要发现更多具有差异化特性的可配体 E3 连接酶可实现全方位的靶向蛋白质降解我们最近利用了化学蛋白质组学平台和分子生物学。鉴定 DCAF16（一种特征较差的 E3 连接酶）作为亲电探针靶标的方法促进蛋白质的核限制性降解，重要的是，DCAF16 能够支持配体- 蛋白质在低参与度（10-40%）下诱导降解，这可能使在该提案中，通过整合来靶向蛋白质，同时最小化其内源底物的干扰。具有细胞生物学和分子生物学方法的化学蛋白质组学平台，我计划 1) 识别内源性 DCAF16的底物和生物学功能，2) 筛选和鉴定破坏癌细胞的小分子通过参与 DCAF16 并诱导新底物降解来促进生长，3) 发现其他 E3 连接酶在不同癌症类型中具有独特且受限的表达，支持配体诱导的靶向蛋白我的长期目标是开发针对 DCAF16 和其他物质的有效且选择性的化学探针。可药物化的 E3 连接酶并开发药理学工具来干预难以药物化甚至“不可药物化”的药物拟议的研究将在顶级的斯克里普斯研究所进行。研究机构结合了尖端的生物和化学研究，提供独特的跨学科研究为了建立成功的职业发展和培训计划，我将参加一系列 TSRI 或 UCSD 主办的课程、讲习班和研讨会，将全面提升我的能力我将掌握指导、教学、科学交流、批判性评估和实验室管理方面的技能。从 Benjamin Cravatt 博士指导的阶段培训中受益匪浅，他拥有 20 多年的临床经验在蛋白质靶点小分子抑制剂以及化学蛋白质组学方面的开发经验技术、拥有癌症转录生物学专业知识的 Michael Erb 博士以及我的建议委员会，其中包括著名科学家杰弗里·凯利博士和戴尔·博格博士拟议的研究。不仅会突破我们对靶向蛋白质降解的认识界限，而且可能是开发针对癌症中重要但不可成药蛋白质的新型疗法的起点。