Determination of P21 upstream signaling in the toxicity of MC and DMC DNA interstrand crosslinks (Student: Melissa Rosas)

总结： 这项“研究补充，以促进多样性， 健康相关的研究”是调查立体异构体的结构之间的关系， 由丝裂霉素C和十氨甲酰丝裂霉素C形成的DNA链间交联（ICL）以及 这些药物的分子机制。丝裂霉素C（MC）是目前用于治疗癌症的抗癌药物。 治疗胃癌、肛门癌和肺癌。MC major ICL的C1“位立体化学构型 是R（α-ICL）。相反，十氨甲酰丝裂霉素C（DMC），一种缺乏O 10的MC衍生物， 氨基甲酰基，生成S立体异构体ICL（β-ICL）。科学的前提是 ICL构成了细胞毒性作用的分子基础， 丝裂霉素核心假设是α和β-DNA局部结构的差异， ICL负责由MC和DMC触发的不同生化反应。在 特别地，与MC相反，DMC处理产生的DNA加合物（DMS-ICL）迅速活化 p53非依赖性细胞死亡途径。因此，研究MC-DMC提供了一个理想的模型， 鉴定在存在或不存在所述抗体的情况下决定细胞信号传导结果的结构特征， 缺乏功能性p53通路。由于p53肿瘤抑制基因在肿瘤细胞中经常发生突变， 人类癌症，需要确定诱导细胞死亡或细胞周期停滞的药物和途径 P53的独立性值得关注。在父项目的范围内， 将利用促进健康相关研究多样性的补充材料来培训候选人， Melissa罗萨斯，以：1：合成含有α/β ICL的寡核苷酸; 2：用 α/β ICL和提取蛋白; 3：上游信号分子，如ATR/BRCA 1， 参与由α或β ICL的存在触发的p21信号通路。梅丽莎的 长期目标是成为临床病理学家。拟议的培训和活动将得到改善 梅丽莎的机会，通过培养关键的技术和专业技能进入医学院。 随着梅丽莎获得新的研究环境，她会变得更加舒适 与其他科学家交流并分享她的工作。这种增强的信心将是一种资产 在医学院的面试和演讲中。为了进一步提高梅丽莎的 竞争力获得医疗计划，她将提出她的研究成果， 会议并发表至少一份手稿。她还将获得职业发展， Edgardo Sanabria-Valentin博士，PRISM项目副主任。作为健康前的职业 顾问，他将协助梅丽莎考虑长期职业目标和所需的技能， 在准备医学院的申请材料时也要做到这一点。

Summary: The overarching goal of the parent proposal for this “research supplement to promote diversity in health-related research” 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. 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. Within the scope of the parent project, this supplement to promote diversity in health-related research will be used to train the candidate, Melissa Rosas, to: 1: Synthesize oligonucleotides containing α/β ICLs; 2: Transfect cells with the α/β ICLs and extract proteins; 3: Validate upstream signaling molecules, such as ATR/BRCA1, involved in the p21 signaling pathway triggered by the presence of either the α or β ICL. Melissa’s long-term goal is to become a clinical pathologist. The training and activities proposed will improve Melissa’s chances to access medical schools by fostering crucial technical and professional skills. As Melissa gains new exposure to the research environment, she will become more comfortable interacting with other scientists and sharing her work. This increased confidence will be an asset during medical school interviews and presentations. In order to further enhance Melissa’s competitiveness to access medical programs, she will present her research findings at conferences and publish at least one manuscript. She will also receive career advisement from Dr Edgardo Sanabria-Valentin, the PRISM Associate Program Director. As the Pre-Health Career Advisor, he will assist Melissa in considering long-term career goals and the skills needed to achieve them as well as in preparing her medical school application material.