Determination of P21 downstream signaling in the toxicity of MC and DMC DNA interstrand crosslinks (Student: Kameza Harun)

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， Kameza Harun：1：合成含有α/β ICL的寡核苷酸; 2：用α/β ICL转染细胞 α/β ICLs并提取蛋白质; 3：纯化下游信号分子，如细胞周期蛋白 B1/Cdk 1，参与p21信号通路，由α或β- ICL。Kameza的长期目标是成为一名肿瘤学临床研究员。培训和 拟议的活动将通过培养 关键的技术和专业技能。随着卡米扎对这项研究有了新的了解 环境，她将变得更加舒适与其他科学家互动，分享她的经验。 工作她将为加入一个研究实验室做好充分的准备。为了进一步加强卡米扎的 竞争力，她将在会议上提出她的研究成果，并发表至少一个 手稿。她还将获得Edgardo Sanabria-Valentin博士的职业荣誉， PRISM副项目主任和健康前职业顾问。他将协助卡米扎 考虑长期职业目标和实现这些目标所需的技能。

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, Kameza Harun to: 1: Synthesize oligonucleotides containing α/β ICLs; 2: Transfect cells with the α/β ICLs and extract proteins; 3: Validate the downstream signaling molecules, such as cyclin B1/Cdk1, involved in the p21 signaling pathway triggered by the presence of either the α or β- ICL. Kameza’s long-term goal is to become a clinical researcher in oncology. The training and activities proposed will improve Kameza’s chances to access MD-PHD programs by fostering crucial technical and professional skills. As Kameza gains new exposure to the research environment, she will become more comfortable interacting with other scientists and sharing her work. She will be fully prepared to join a research laboratory. In order to further enhance Kameza’s competitiveness, 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 and Pre-Health Career Advisor. He will assist Kameza in considering long-term career goals and the skills needed to achieve them.