Identification of critical cellular pathways triggered by mitomycins interstrand crosslinks

丝裂霉素链间交联触发的关键细胞途径的鉴定

基本信息

批准号：
10629504
负责人：
Elise Champeil
金额：
$ 15.9万
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：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10629504
关键词：
Antineoplastic Agents Apoptotic Attention Biochemical Bioinformatics Biological Biological Assay Biomimetics C10 CHEK1 gene Cell Cycle Cell Cycle Arrest Cell Death Cell Death Induction Cell Line Cells Clinic Clinical Complex DNA DNA Adduction DNA Adducts DNA Interstrand Crosslinking DNA Structure Development Drug usage G1 Phase Goals Human K-562 Knowledge Label Laboratories MCF7 cell Malignant Neoplasms Mammalian Cell Mediating Methods Minor Groove Mitomycin C Mitomycins Modeling Modernization Modification Molecular Molecular Conformation Mutate Mutation Outcome Pathway interactions Pharmaceutical Preparations Pharmacotherapy Phosphorylation Positioning Attribute Post-Translational Protein Processing Primary Lesion Process Proteins Proteomics Research Route Signal Transduction Stereoisomer Structure TP53 gene Transfection adduct analog cancer cell cancer therapy chemotherapeutic agent crosslink cytotoxic cytotoxicity experimental study genetic makeup individualized medicine interest mutant novel personalized medicine protein expression response transcription factor tumor

项目摘要

This proposal’s objective is to identify critical cellular pathways triggered by the presence of DNA interstrand crosslinks (ICLs) produced by mitomycins, and to identify how the structure of mitomycins ICLs determines the cellular signaling leading to cell death/cell cycle arrest in the absence of a functioning p53. This study includes mitomycin C (MC), an anti-cancer drug currently used in the clinics, decarbamoyl mitomycin C (DMC), a derivative of mitomycinc (MC) and a novel MC-derivative, MC-Lex, synthesized in our laboratory. These compounds form highly cytotoxic interstrand crosslinks (ICLs) that share common structural features. MC forms the α-ICL with DNA, DMC the β-ICL and MC-Lex the γ-ICL. Representative conformations of the three ICLs indicate that the level of DNA perturbation induced by each ICL increases in the order α<β<γ, with the α-ICL inducing minimal DNA perturbation and the γ-ICL shows significant DNA distortion and widening of the minor groove. Previous research has shown that the three drugs (MC, DMC, MC-Lex) also differ in their cytotoxicity and p53 signaling. Since ICLs are the lesions primarily responsible for the cytotoxicity of mitomycins, this indicates that the three ICLs could trigger very diverse biochemical cellular mechanisms, mediated by specific mutations present in cancer cells, a hallmark of modern cancer therapy. The proposed research will establish how the ICLs behave as biological signals to trigger different cell death/cell cycle arrest pathways. Knowledge of how differences in the modification of the local DNA structure by mitomycins correlates with their cytotoxicity is crucial for understanding the action of clinically used drugs. Therefore, the study of these three structurally different ICLs (α/β/γ ICLs) provides an ideal model for identifying structural features determining the cell-signaling outcome in the presence or the absence of a functioning p53 pathway. Since p53 tumor suppressor is mutated in more than 50% of human cancers, the need to identify cellular pathways that induce cell death or cell cycle arrest independently of p53 deserves substantial attention. This has the potential to identify treatments that could be tailored to cancer cells with mutations in the p53 gene for personalized treatment consideration based on the genetic makeup of a tumor. To correlate ICL structures with signaling outcome, we will be pursue the following aims: Specific aim #1: Synthesis of MC, DMC and MC-Lex ICLs (α,β,γ-ICLs) using biomimetic methods. Specific aim #2: Determination of p53-dependent and independent DNA adducts (ICLs) response mechanisms using quantitative label free proteomics and bioinformatics Specific aim #3: Mechanistic of CHK1 mediated cell cycle arrest triggered by MC/DMC and α/β-ICL Expected outcomes: This study will identify structural ICL features that determine the cell signaling outcome in the presence or the absence of a functioning p53 pathway, a feature of particular interest for the development of personalized anticancer therapy.

该提案的目标是确定由 DNA 链间存在触发的关键细胞途径丝裂霉素产生的交联 (ICL)，并确定丝裂霉素 ICL 的结构如何决定在缺乏功能性 p53 的情况下，细胞信号传导导致细胞死亡/细胞周期停滞。包括目前临床上使用的抗癌药物丝裂霉素C（MC）、去氨甲酰丝裂霉素C (DMC) 是丝裂霉素 (MC) 的衍生物，也是我们实验室合成的新型 MC 衍生物 MC-Lex。这些化合物形成具有共同结构特征的高细胞毒性链间交联 (ICL)。 MC 与 DNA 形成 α-ICL，DMC 形成 β-ICL，MC-Lex 形成 γ-ICL。三个 ICL 表明每个 ICL 引起的 DNA 扰动水平按 α<β<γ 的顺序增加，其中 α-ICL 引起最小的 DNA 扰动，而 γ-ICL 显示出显着的 DNA 扭曲和加宽先前的研究表明，这三种药物（MC、DMC、MC-Lex）的差异也在于。它们的细胞毒性和 p53 信号传导，因为 ICL 是主要负责细胞毒性的病变。丝裂霉素，这表明三种 ICL 可以触发非常不同的生化细胞机制，由癌细胞中存在的特定突变介导，这是现代癌症治疗的标志。研究将确定 ICL 如何作为生物信号触发不同的细胞死亡/细胞周期停滞了解丝裂霉素对局部 DNA 结构的修饰有何差异与其细胞毒性的相关性对于了解临床使用药物的作用至关重要。对这三种结构不同的 ICL（α/β/γ ICL）的研究为识别结构提供了理想的模型决定在存在或不存在功能性 p53 通路的情况下细胞信号转导结果的特征。由于 p53 肿瘤抑制基因在超过 50% 的人类癌症中发生突变，因此需要鉴定细胞独立于 p53 诱导细胞死亡或细胞周期停滞的途径值得大量关注。有潜力确定针对 p53 基因突变的癌细胞的治疗方法根据肿瘤的基因组成进行个性化治疗考虑，以关联 ICL 结构。随着取得的成果，我们将追求以下目标：具体目标#1：使用仿生方法合成 MC、DMC 和 MC-Lex ICL（α,β,γ-ICL）。具体目标#2：确定 p53 依赖性和独立 DNA 加合物 (ICL) 反应使用定量无标记蛋白质组学和生物信息学的机制具体目标#3：MC/DMC 和 α/β-ICL 触发 CHK1 介导的细胞周期停滞的机制预期结果：本研究将确定决定细胞信号转导结果的 ICL 结构特征在存在或不存在功能性 p53 通路的情况下，这是一个特别令人感兴趣的特征个性化抗癌治疗的发展。