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.

摘要： 这份《促进多样性的研究补充文件》的主要目标是 与健康相关的研究“是考察立体异构体的结构之间的关系 丝裂霉素C和脱氨酰丝裂霉素C形成的DNA链间交联物 这些药物的分子机制。丝裂霉素C(MC)是一种抗癌药物，目前用于 治疗胃癌、肛门癌和肺癌。MC大分子ICL的C_1‘’立体构型 为R(α-icl)。相比之下，脱氨甲酰丝裂霉素C(DMC)是MC的衍生物，缺乏O10 氨甲酰基，生成S立体异构体ICl(β-ICl)。科学的前提是 建议的研究是ICL构成了细胞毒作用的分子基础 丝裂霉素类。中心假设是α和β局部dna结构的差异- ICL负责MC和DMC触发的不同生化反应。在……里面 特别是，与MC相反，由DMC处理产生的dna加合物(-icl)迅速激活 一条不依赖于p53的细胞死亡途径。因此，MC-DMC的研究为下一步的研究提供了理想的模型 确定在存在或存在的情况下决定细胞信号结果的结构特征 缺乏功能正常的P53途径。由于P53肿瘤抑制基因经常在 人类癌症，需要识别导致细胞死亡或细胞周期停滞的药物和途径 与P53无关的问题值得大力关注。在父项目的范围内，这 用于促进健康相关研究多样性的补充将用于培训候选人， 克里斯蒂娜·冈萨雷斯：1：合成含有α/βICL的寡核苷酸；2：用 这些寡核苷酸；3：进行RNA提取，以确定基因表达的变化 转染α或β-ICL后细胞反应的特征；4：确定改变 微RNA(MiRNA)在细胞对MC/DMC暴露的反应中的分布；5：确定这些 药物影响功能性miRNAs以及是否形成MC/DMC-miRNAs加合物；以及6：获得技能 在转录组数据的生物信息学分析中。克里斯蒂娜的长期目标是成为一名医生 研究科学家。拟议的培训和活动将增加克里斯蒂娜获得 通过培养关键的技术和专业技能，开展医学博士项目。以进一步增强 克里斯蒂娜的竞争力，她将与我们的合作者桑切斯-维加博士密切合作，负责人 纪念斯隆·凯特琳癌症中心结直肠癌计算生物部 癌症科。她还将接受Edgardo Sanabria-Valentin博士的职业建议 PRISM副项目主任兼健康前职业顾问。