p53-Independent Cell Death Signaling by Mitomycin DNA Adducts

丝裂霉素 DNA 加合物的 p53 独立细胞死亡信号传导

• 批准号: 7427239
• 负责人: Jill E. Bargonetti
• 金额: $ 32.9万
• 依托单位: HUNTER COLLEGE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-06 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7427239
• 关键词: Address; Alkylating Agents; Antibodies; Antineoplastic Agents; Apoptotic; Attention; Autophagocytosis; Biological Models; Caenorhabditis elegans; Cancer cell line; Cell Death; Cell Death Signaling Process; Cell Line; Cells; Cessation of life; Classification; Complex; Cultured Cells; DNA; DNA Adducts; DNA Damage; DNA Repair; DNA repair protein; Dependency; Development; Drug Delivery Systems; Drug Design; Funding; Genes; Genetic; Genotype; Germ Cells; Goals; Hereditary Disease; Homologous Gene; Human; Human Cell Line; In Vitro; Knock-out; Knowledge; Laboratories; Laboratory Research; Lesion; Longevity; Malignant Neoplasms; Mediating; Meiosis; Methods; Mitomycin; Mitomycins; Mitotic; Modeling; Mutation; Necrosis; Nematoda; Nuclear Protein; Nuclear Proteins; Nucleosides; Oligonucleotides; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Phosphorylation; Phosphotransferases; Play; Positioning Attribute; Pre-Clinical Model; Protein Binding; Protein p53; Proteins; Public Health; Publishing; RNA Interference; Recruitment Activity; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site; Staging; System; TP53 gene; Tissue Model; Work; adduct; cancer cell; cancer therapy; cell killing; chromatin immunoprecipitation; colon cancer cell line; comparative; cytotoxicity; design; drug development; in vivo; independency; innovation; model development; mutant; plasmid DNA; protein expression; research study; sensor; stereochemistry; tumor

DESCRIPTION (provided by applicant): The need to identify drugs and pathways that induce cell death independently of p53 deserves substantial attention. Induction of cancer cell death is challenging when p53 and other apoptotic pathways are inactivated, and requires chemotherapeutics that induce cell death pathways that do not require the tumor suppressor p53, as at least 50 percent of cancers lack a functional p53 pathway. We published that different mitomycins, which are alkylating agents, activate p53-dependent and p53-independent cell death. We see the mitomycin derivative 10-decarbamyl mitomycin C (DMC), as opposed to mitomycin C (MC), can rapidly activate a p53-independent cell death pathway which lacks markers of apoptotic death. DMC-induced cell death is a good model for identifying targets for the induction of p53-independent cell death pathways, such as necrosis and autophagy. Our central hypothesis is that DMC provokes p53-independent cell death due to specific DNA adducts formed and that elucidating the pathway(s) will enable efficient drug design parameters. Identifying the mechanisms by which DMC induces cell death is the focus of this proposal and will be a first step toward developing drugs to strategically induce cell death for the many cancers lacking functional p53 pathways. Despite the importance of p53-independent cell death, little is known about the activation of such pathways. Four aims are proposed that will use a human cancer cell line with inducible p53 and its isogenic pair (without p53) to examine p53 dependency and independency in cell death signaling, and C. elegans to serve as a preclinical model system. (1) Identify the human signal transduction pathways required for cell death in the presence and absence of p53 that are activated and/or repressed by DMC and MC. (2) Analyze the ability of DMC and MC DNA adducts in vitro and in vivo to recruit DNA repair proteins in the presence and absence of p53. (3) Perform comparative analysis of the influence of the p53 pathway and the autophagy pathway on the chemotherapeutic DNA damage-induced cell death in C. elegans and human cell systems. (4) Obtain R01 funding for my laboratory research. There is a gap in knowledge as to how to target for p53-independent cell death. Knowledge of the signaling mechanisms for p53-independent cell death pathways will help in targeting drugs to patients who have cancers that lack functional p53.

描述（由申请人提供）：需要鉴定独立于p53诱导细胞死亡的药物和途径，值得高度重视。当p53和其他凋亡通路失活时，诱导癌细胞死亡是具有挑战性的，并且需要诱导不需要肿瘤抑制因子p53的细胞死亡通路的化疗药物，因为至少50%的癌症缺乏功能性p53通路。我们发表了不同的丝裂霉素，它们是烷基化剂，激活p53依赖性和p53非依赖性细胞死亡。我们发现丝裂霉素衍生物10-decarbamyl丝裂霉素C (DMC)，与丝裂霉素C （MC）相反，可以快速激活缺乏凋亡死亡标志物的p53非依赖性细胞死亡途径。dmc诱导的细胞死亡是一个很好的模型，用于识别诱导p53非依赖性细胞死亡途径（如坏死和自噬）的靶点。我们的中心假设是，由于形成了特定的DNA加合物，DMC引发了p53非依赖性细胞死亡，阐明这一途径将使有效的药物设计参数成为可能。确定DMC诱导细胞死亡的机制是本提案的重点，并且将是开发药物以战略性地诱导许多缺乏功能性p53通路的癌症细胞死亡的第一步。尽管p53非依赖性细胞死亡很重要，但对这些途径的激活知之甚少。研究人员提出了四个目标，将使用具有诱导型p53及其等基因对（不含p53）的人类癌细胞系来检测p53在细胞死亡信号传导中的依赖性和独立性，并将秀丽隐杆线虫作为临床前模型系统。(1)确定在p53存在和缺失的情况下，细胞死亡所需的人类信号转导通路被DMC和MC激活和/或抑制。(2)分析在p53存在和缺失的情况下，DMC和MC DNA加合物在体外和体内募集DNA修复蛋白的能力。(3)比较分析p53通路和自噬通路对秀丽隐杆线虫和人类细胞系统化疗性DNA损伤诱导的细胞死亡的影响。(4)获得实验室研究R01经费。