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

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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.