Differences in RNA and miRNA expression in response to MC and DMC stereoisomeric interstrand crosslinks (Student: Christina Gonzalez)

MC 和 DMC 立体异构体链间交联反应中 RNA 和 miRNA 表达的差异（学生：Christina Gonzalez）

• 批准号: 10544084
• 负责人: Elise Champeil
• 金额: $ 9.61万
• 依托单位: JOHN JAY COLLEGE OF CRIMINAL JUSTICE
• 依托单位国家: 美国
• 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/10544084
• 关键词: Antineoplastic Agents; Apoptotic; Attention; Biochemical; Bioinformatics; Cell Cycle Arrest; Cell Death; Cells; Colorectal Cancer; Computational Biology; DNA Adducts; DNA Interstrand Crosslinking; DNA Structure; Data; Fostering; Gene Expression; Goals; Head; Health; Human; Malignant Neoplasms; Malignant neoplasm of anus; Malignant neoplasm of lung; Medical Research; Memorial Sloan-Kettering Cancer Center; MicroRNAs; Mitomycin C; Mitomycins; Modeling; Molecular; Mutate; Oligonucleotides; Outcome; Parents; Pathway interactions; Pharmaceutical Preparations; RNA; Research; Research Proposals; Scientist; Signal Transduction; Structure; Students; TP53 gene; Training; Training Activity; Transfection; Work; adduct; base; career; chemotherapeutic agent; crosslink; cytotoxic; genetic makeup; improved; malignant stomach neoplasm; parent project; personalized medicine; programs; response; skills; transcription factor; transcriptomics; tumor

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, Christina Gonzalez to: 1: Synthesize oligonucleotides containing α/β ICLs; 2: Transfect cells with these oligonucleotides; 3: Perform RNA extraction to determine the changes to gene expression profiles in the cellular response after transfection of the α or β-ICL; 4: Determine changes to microRNA (miRNA) profiles in the cellular response to MC/DMC exposure; 5: Identify how these drugs impact functional miRNAs and if MC/DMC-miRNAs adducts are formed; and 6: Gain skills in bioinformatic analysis of transcriptomics data. Christina’s long-term goal is to become a medical research scientist. The training and activities proposed will improve Christina’s chances to access MD-PHD programs by fostering crucial technical and professional skills. To further enhance Christina’s competitiveness, she will work closely with our collaborator, Dr Sanchez-Vega, head of the Computational Biology Division at Memorial Sloan Kettering Cancer Center Colorectal Cancer Unit. She will also receive career advisement from Dr Edgardo Sanabria-Valentin, the PRISM Associate Program Director and Pre-Health Career Advisor.

概括： 家长提案的总体目标是“促进多样性的研究补充” 健康相关研究”是研究立体异构体结构之间的关系 由丝裂霉素 C 和去氨甲酰丝裂霉素 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药物。 治疗胃癌、肛门癌和肺癌 MC 主要 ICL 的 C1'' 立体化学构型。 是 R (α-ICL)，相反，去氨甲酰丝裂霉素 C (DMC)，是缺少 O10 的 MC 衍生物。 氨基甲酰基，生成 S 立体异构 ICL (β-ICL) 的科学前提。 拟议的研究是 ICL 构成了细胞毒性作用的分子基础 丝裂霉素的中心假设是α和β-的局部DNA结构的差异。 ICL 负责 MC 和 DMC 触发的不同生化反应。 特别是，与 MC 相反，DMC 处理（-ICL）产生的 DNA 加合物迅速激活 因此，MC-DMC 研究提供了一个理想的模型。 结构特征决定细胞信号转导结果的存在或 由于 p53 肿瘤抑制因子经常发生突变，因此缺乏有效的 p53 通路。 人类癌症，需要识别诱导细胞死亡或细胞周期停滞的药物和途径 在父项目的范围内，独立于 p53 值得大量关注。 促进健康相关研究多样性的补充品将用于培训候选人， Christina Gonzalez：1：合成含有 α/β ICL 的寡核苷酸；2：用 3：进行RNA提取以确定基因表达的变化 转染 α 或 β-ICL 后的细胞反应概况 4：确定变化； MC/DMC 暴露的细胞反应中的 microRNA (miRNA) 概况 5：确定这些是如何发生的； 如果形成 MC/DMC-miRNA 加合物，则会影响功能性 miRNA 和药物；6：获得技能； 克里斯蒂娜的长期目标是成为一名医学博士。 拟议的培训和活动将增加克里斯蒂娜获得研究科学家的机会。 MD-PHD 课程通过培养关键的技术和专业技能来进一步提高。 克里斯蒂娜的竞争力，她将与我们的合作者，负责人桑切斯-维加博士密切合作 纪念斯隆凯特琳癌症中心结直肠癌计算生物学部 她还将获得癌症中心埃德加多·萨纳布里亚·瓦伦丁 (Edgardo Sanabria-Valentin) 博士的职业建议。 PRISM 副项目总监和健康前职业顾问。