Role of p21 in the toxicity of MC and DMC DNA Interstrand Crosslinks

p21 在 MC 和 DMC DNA 链间交联毒性中的作用

基本信息

批准号：
10377556
负责人：
Elise Champeil
金额：
$ 12.03万
依托单位：
JOHN JAY COLLEGE OF CRIMINAL JUSTICE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10377556
关键词：
Antineoplastic Agents Apoptotic Attention BRCA1 gene Biochemical Biological Biomimetics Cell Cycle Arrest Cell Death Cell Line Cells Co-Immunoprecipitations Complement Complex DNA Adducts DNA Interstrand Crosslinking DNA Structure DNA crosslink Data Goals Human K-562 Laboratories M cell MCF7 cell Malignant Neoplasms Malignant neoplasm of anus Malignant neoplasm of lung Mediating Methods Mitomycin C Mitomycins Modeling Molecular Monitor Mutate Outcome Pathway interactions Pharmaceutical Preparations Post-Translational Protein Processing Proteomics Protocols documentation Research Role Route Signal Pathway Signal Transduction Structure Structure-Activity Relationship TP53 gene Toxic effect Western Blotting Work adduct base cancer cell chemotherapeutic agent crosslink cyclin B1 cytotoxic functional status genetic makeup innovation malignant stomach neoplasm molecular targeted therapies novel personalized medicine protein expression response transcription factor tumor

项目摘要

Abstract The overarching goal of this proposal is to investigate the relationship between the structure of stereoisomeric DNA Interstrand Crosslinks (ICLs) formed by Mitomycin C and Decarbamoylmitomycin C and the molecular mechanisms of these drugs. Mitomycin C (MC) is an anticancer drug currently used to treat stomach, anal and lung cancers. The stereochemical configuration at C1’’ of MC major ICL is R (α ICL). In contrast, Decarbamoylmitomycin C (DMC), a derivative of MC lacking the O10 carbamoyl group, generates the S stereoisomeric ICL (β ICL). The scientific premise of the proposed research is that ICLs constitute the molecular basis for the cytotoxic effects of mitomycins. The central hypothesis is that differences in the local DNA structures of the α and β-ICLs are responsible for the distinct biochemical responses triggered by MC and DMC. In particular, contrary to MC, the DNA-adducts generated by DMC treatment (-ICL) rapidly activate a p53-independent cell death pathway. Thus, the study MC- DMC provides an ideal model for identifying structural features determining the cell signaling outcome in the presence or the absence of a functioning p53 pathway. The significance of this project lies in determining the structure-activity relationship for stereoisomeric DNA crosslink adducts. In addition, the proposed research will establish how the α and β-ICLs behave as biological signals to different cell death pathways. The key innovation of this project is to generate stereoisomeric ICLs on a scale which will allow the study of biochemical responses using our newly developed biomimetic method. Finally, since p53 tumor suppressor is frequently mutated in human cancers, the need to identify drugs and pathways that induce cell death or cell cycle arrest independently of p53 deserves substantial attention. In order to correlate MC and DMC-adducts structures with the toxicity of the α-ICL and β-ICL, the following three aims will be pursued: 1) Synthesis of MC and DMC DNA adducts. 2) Determination of p53-dependent and independent MC/DMC DNA adducts response mechanisms using proteomics. 3) Determination of the molecular signaling pathway involved the in G2/M cell cycle arrest triggered by MC/DMC DNA-adducts. Our study will reveal molecular and cellular networks up or downregulated by MC/DMC and the α/β-ICLs in p53-dependent and p53-independent cell lines. The proposed work will therefore identify novel critical molecular therapeutic targets of MC and DMC.

抽象的该提案的总体目标是研究结构之间的关系由丝裂霉素 C 和丝裂霉素形成的立体异构 DNA 链间交联 (ICL) 去氨甲酰丝裂霉素 C 和这些药物的分子机制是丝裂霉素 C (MC)。目前用于治疗胃癌、肛门癌和肺癌的抗癌药物。 MC 主要 ICL 的 C1'' 构型是 R (α ICL)，相比之下，去氨甲酰丝裂霉素 C (DMC)，缺少 O10 氨基甲酰基的 MC 衍生物，生成 S 立体异构体 ICL（β ICL）所提出的研究的科学前提是 ICL 构成了分子基础。对于丝裂霉素的细胞毒性作用的中心假设是局部差异。 α 和 β-ICL 的 DNA 结构负责不同的生化反应由 MC 和 DMC 触发，特别是与 MC 相反，DMC 产生的 DNA 加合物。治疗 (-ICL) 迅速激活 p53 独立的细胞死亡途径，因此，研究 MC-。 DMC 提供了一个理想的模型来识别决定细胞信号传导的结构特征存在或不存在功能性 p53 通路时的结果。该项目的意义在于确定了结构-活性关系此外，拟议的研究将确定如何实现立体异构 DNA 交联加合物。 α 和 β-ICL 充当不同细胞死亡途径的生物信号关键创新。该项目的目的是生成一定规模的立体异构 ICL，以便研究最后，使用我们新开发的仿生方法进行生化反应。抑制基因在人类癌症中经常发生突变，需要确定药物和途径独立于 p53 诱导细胞死亡或细胞周期停滞值得大量关注。为了将 MC 和 DMC 加合物结构与 α-ICL 和 β-ICL 的毒性关联起来，我们将追求以下三个目标：1）MC和DMC DNA加合物的合成。确定 p53 依赖性和独立的 MC/DMC DNA 加合物反应使用蛋白质组学的机制3）确定所涉及的分子信号通路。由 MC/DMC DNA 加合物触发的 G2/M 细胞周期停滞。我们的研究将揭示 MC/DMC 上调或下调的分子和细胞网络以及因此，拟议的工作将在 p53 依赖性和 p53 独立细胞系中进行。确定 MC 和 DMC 的新的关键分子治疗靶点。