靶向泛素E3连接酶的抗肿瘤免疫调节剂的新底物和新机理研究

Targeting disease specific protein degradation via ubiquitin-proteasome system (UPS) is an important approach for cancer therapy. Recently, the molecular mechanism of immunomodulatory imide drugs (IMiDs), such as thalidomide, in multiple myeloma has been demonstrated to be its direct binding to CRBN, part of an ubiquitin E3 ligase complex, for specific substrate degradation, which is an important conceptual advancement in anti-cancer drug development. Nevertheless, in another important indication of IMiDs, non-Hodgkin's lymphoma, their specific degradation substrates and the underlying molecular mechanisms of action and drug resistance still remain unclear. In this proposal, by taking the unique advantage of mass spectrometry-based proteomics technologies in protein degradation and mechanism study, we primarily focus on the systematic characterization of novel degradation substrates of three important IMiDs, lenalidomide, CC-122 and CC-885, in non-Hodgkin's lymphoma. We will carry out the quantitive proteomics, ubiquitinome and drug interactome analyses to comprehensively identify degradation substrates of these drugs. Together with bioinformatic analysis, and biochemical and pharmacological methods, we will further validate the degradation substrates identified in our proteomics study, elucidate their downstream molecular functions and drug resistance mechanism, and identify potential biomarkers indicative of their therapeutic efficacy. This study will provide a systematic view and a basis on the molecular mechanism of targeting the CRBN E3 ligase and overcoming drug resistance for more effective therapy in non-Hodgkin's lymphoma.

靶向泛素-蛋白酶体系统调控疾病特异性蛋白降解，是肿瘤治疗的重要策略。近期研究证明，沙利度胺类免疫调节剂，在治疗多发性骨髓瘤等疾病中的机制，是通过靶向CRBN E3泛素连接酶，调控特异性蛋白底物降解而起效，是近年来抗肿瘤药物研究的重要概念性突破。本项目将针对此类药物在其重要适应症非霍奇金淋巴瘤中降解底物特异性仍不明确、分子机制不清以及耐药问题，利用基于质谱的蛋白质组学技术在研究蛋白降解底物和机制中的独特优势，运用定量蛋白质组学、泛素化修饰组学、基于分子探针的药物相互作用蛋白组分析，结合生物信息学，以及生化和分子药理学等技术手段，系统鉴定和确证此类药物(包括来那度胺、CC-122和CC-885)在非霍奇金淋巴瘤中的特异性降解底物，阐述其分子调控新机制和耐药新机理，鉴定此类药物疗效的潜在生物标志物，为靶向CRBN泛素连接酶系统类药物的分子机理阐明，克服耐药和精准用药研究，提供全局性的视角和依据。

靶向泛素-蛋白酶体系统调控疾病特异性蛋白质降解，是肿瘤治疗的重要策略。本项目利用基于质谱的蛋白质组学技术、泛素化修饰组学、基于分子探针的药物相互作用蛋白组分析等，系统鉴定和确证靶向泛素-蛋白酶体系统类药物的分子机理，为疾病精准用药研究提供全局性的视角和依据。在本项目的支持下，我们1）揭示了度胺类靶向蛋白降解剂CC-885诱导蛋白降解的新底物及其作用机制；2）揭示靶向蛋白降解剂E7070诱导蛋白降解的作用底物及其潜在抗肿瘤机制；3）实现人类肺癌多维分子全景图谱解析、分子分型及潜在生物标志物发现；4）建立KRAS突变肿瘤分子分型和靶向磷酸化的个性化治疗新策略；5）揭示AMPK介导磷酸化在DNA损伤反应中的新型分子机制；6）解析了组蛋白去乙酰化酶抑制剂肿瘤耐药新机制并发现了克服组蛋白去乙酰化酶抑制剂肿瘤耐药的潜在药物联用新策略。此外，在我们的拓展工作中，我们进行了蛋白质组学技术方法学的开发及其它类型药物的作用机制研究。我们开发了蛋白质翻译后修饰大规模数据分析程序PTMiner，以及质谱数据采集新方法BoxCar，并对干扰素等药物作用机制进行了深度解析。整体来讲，我们的研究成果不仅促进了对泛素-蛋白酶体系统与疾病调控的机制认知，而且为疾病的精准治疗提供了重要资源和线索。