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

基本信息

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
项目类别：
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

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

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项目摘要

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.

描述（由适用提供）：顺铂和卡铂（白金药物）是最成功的抗肿瘤药物之一，用于治疗有证明的，乳房，卵巢，膀胱，膀胱，颈部，颈部和肺癌。与许多抗肿瘤药物一样，治疗的有效性可能因患者而异，并且抗药性的发生是一个重大问题。铂的治疗效应是基于与在一或两个完整的DNA链上的两个购买碱基的反应中形成不同类型的DNA加合物（主要是链间交联）的形成。铂处理的耐药性是通过多种过程的上调或下调（包括金属输入和流入细胞），细胞内代谢以及通过DNA修复途径去除顺铂DNA加合物的。 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从治疗和保留可能具有抵抗力的严重副作用相关的治疗方法。我们建议使用该R21应用，以合成含有1,2GG-，1,2-AG-，1,3-GNG-Intrastrand和1,2-GC Interstrand Platinum Crosslinks的寡核苷酸，将它们与KLH载体蛋白相结合，并使用它们与klh carrier蛋白相比，并将其与klh型klh蛋白质相关，以识别单个抗元素的抗生素。然后将在具有特定DNA修复缺陷的细胞系中进一步验证这些单克隆抗体。我们预测，我们将测量NCI-60收集的乳腺癌和卵巢癌细胞系中的铂水平，这些水平的特征是药物敏感性或耐药性。我们将确定在铂反应性细胞系中特定的ADDCT或所有加合物的水平是否升高。相反，我们将测试在耐药细胞系中是否没有特定的加合物。我们预计，在这里提出的研究完成后，我们将开发一系列独特的试剂，这些试剂将对研究和医学界有很大的用途来确定哪些铂金合并在临床上是最相关的，并致力于开发一种可靠的方法来预测铂金处理的临床结果。