Generation and characterization of adduct-specific anti cisplatin DNA antibodies

加合物特异性抗顺铂 DNA 抗体的生成和表征

• 批准号: 8951743
• 负责人: Orlando D. Scharer
• 金额: $ 20.41万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8951743
• 关键词: Address; Adverse effects; Affect; Antibodies; Antibody Specificity; Antigens; BRCA2 gene; Basic Science; Binding; Biological Assay; Biopsy; Breast; Cancer cell line; Carboplatin; Carrier Proteins; Cell Line; Cells; Chemotherapy-Oncologic Procedure; Cisplatin; Clinic; Clinical; Collection; Communities; Cytotoxic Chemotherapy; Cytotoxic agent; DNA; DNA Adducts; DNA Damage; DNA Repair; DNA Repair Pathway; Defect; Down-Regulation; Drug Combinations; Drug Targeting; Drug resistance; Drug toxicity; Drug usage; ERCC1 gene; Excision; Future; Generations; Genome; Goals; Hybridomas; Individual; Keyhole Limpet Hemocyanin; Link; Lung; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Malignant neoplasm of lung; Malignant neoplasm of ovary; Malignant neoplasm of testis; Malignant neoplasm of urinary bladder; Measurement; Measures; Medical Research; Metabolism; Metals; Methods; Monitor; Monoclonal Antibodies; Mus; Neck Cancer; Nucleotide Excision Repair; Oligonucleotides; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Platinum; Platinum adduct; Point Mutation; Process; Property; Purines; Reaction; Reagent; Resistance; Sampling; Site; Specificity; Testing; Therapeutic; Therapeutic Effect; Toxic effect; Treatment Efficacy; Tumor Cell Line; Up-Regulation; Validation; Work; XRCC3 gene; Xeroderma Pigmentosum Complementation Group A; Xeroderma Pigmentosum Complementation Group G; adduct; antitumor agent; antitumor drug; base; cisplatin-DNA adduct; crosslink; cytotoxic; drug metabolism; drug sensitivity; endonuclease; inhibitor/antagonist; insight; interest; malignant breast neoplasm; oncology; public health relevance; purine; repaired; resistance mechanism; response; targeted treatment; tumor; tumor DNA; ultraviolet irradiation

 DESCRIPTION (provided by applicant): Cisplatin and carboplatin (platinum drugs) are among the most successful antitumor drugs and are used to treat testicular, breast, ovarian, bladder, neck and lung cancer. As with many antitumor agents, the efficacy of treatment can vary greatly from patient to patient and the occurrence of resistance is a significant problem. The therapeutic effect of the platinums is based on the formation of different types of DNA adducts, primarily intra- and interstrand crosslinks, by the reaction with two purine bases on one or two complementary strands of DNA. The resistance to platinum treatment occurs by a the up- or down-regulation of several processes, including metal transport in and out of the cell, intracellular metabolism and the removal of the cisplatin DNA adducts by DNA repair pathways. The primary hypothesis of the work proposed in this application is that by developing a monoclonal antibodies with specificity for the individual platinum DNA adducts in the genomes of cells we will be able to 1) determine which platinum adduct is the most therapeutically relevant one, 2) understand the repair mechanisms responsible for resistance, 3) test the sensitivity or resistance to platinum in tumor biopsies and 4) to select patients that are most likely to benefit from therapy and sparing those that are likely to be resistant the severe side effect associated with this type of treatment. In this R21 application, we propose to synthesize oligonucleotides containing the 1,2-GG-, 1,2-AG-, 1,3-GNG-intrastrand and 1,2-GC-interstrand platinum crosslinks, couple them to the KLH carrier protein and use them to immunize mice to generate monoclonal antibodies that recognize the individual adducts with high specificity. These monoclonal antibodies will be then further validated in cell lines with specific DNA repair defects. We predict that these reagents will enable us to show that platinum intrastrand crosslinks persist in cells with defects in the nucleotide excision repair (NER) pathway, while interstrand crosslinks will persist in cells with defects interstrand crosslink (ICL) repair pathwa. We will then measure platinum levels in breast and ovarian cancer cell lines of the NCI-60 collection, which have been characterized for drug sensitivity or resistance. We will determine whether the levels of a specific adduct or all adduct are elevated in platinum-responsive cell lines. Conversely, we will test whether a specific adduct is absent in resistant cell lines. We expect that upon completion of the studies proposed here we will have developed a set of unique reagents that will be of tremendous use for the research and medical community to determine which platinum adducts are clinically most relevant and to work toward developing a robust method to predict clinical outcomes of platinum treatments.