Enhanced delivery of site-specific DNA damaging toxins to prostate cancercells

增强向前列腺癌细胞输送特定位点 DNA 损伤毒素

基本信息

批准号：
10654187
负责人：
Arthur R Frampton
金额：
$ 44.7万
依托单位：
UNIVERSITY OF NORTH CAROLINA WILMINGTON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-08 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10654187
关键词：
Address Adenine Affinity African American Androgen Receptor Androgens Animal Model Apoptosis Artificial Membranes Binding Biological Assay Biological Process Biomedical Research Breast Cancer Cell Cancer Etiology Cell Death Cell Membrane Permeability Cell Nucleus Cells Chemotherapy-Oncologic Procedure DNA DNA Adduction DNA Adducts DNA Binding DNA Damage DNA Double Strand Break DNA Methylation DNA lesion Detection Development Disease Epithelial Cells Estrogen Receptors Foundations Future Genes Goals High Pressure Liquid Chromatography Homidium Bromide LNCaP Lesion Ligands Malignant neoplasm of prostate Measures Mediating Methylation Minor Groove Molecular Biology Techniques Nature Necrosis Nuclear Nuclear Receptors Organic Synthesis Outcome PC3 cell line Property Prostate Prostate Cancer therapy Reaction Research Risk Role Second Primary Cancers Site Techniques Testing Thymine Toxic effect Toxin Training Underrepresented Minority adduct analog calf thymus DNA cancer cell chemical standard chemotherapy malignant breast neoplasm overexpression prostate cancer cell prostate cancer cell line prostate cancer model racial disparity receptor binding side effect targeted agent tissue culture treatment response undergraduate student

项目摘要

ABSTRACT The overall goal of our research is to develop a dependable strategy for selectively targeting different cancer cells and producing specific types of DNA-damage in them, leading to their targeted destruction. These molecules will help us understand DNA-damage mediated biological processes in cancer cells and lay the foundation for advances in disease treatment. The objective of this R15 proposal is to develop molecules that target prostate cancer (PCa) cells and cause clustered DNA damage within them leading to the formation of lethal DNA double strand breaks (DSBs). This project addresses the current critical need in PCa treatment to develop new chemotherapy agents with higher selectivity and potency but without toxic side-effects. Our strategy is to synthesize molecules that bind to the androgen receptor (AR), which is overexpressed in PCa cells, get escorted to the nucleus of these cells by AR action, and generate predominantly N3-methyladenine (3MeA) DNA adducts in close proximity. Cellular processing of these closely spaced DNA lesions will lead to the formation DNA DSBs. This approach is partially based upon our successful targeted destruction of breast cancer cells using molecules that bind to estrogen receptors (ERs) which are overexpressed in many breast cancers. We hypothesize that our strategy will lead to the targeted, potent destruction of PCa cells due to the lethal nature of DNA DSBs and because the AR and ER belong to the same superfamily of nuclear receptors and function similarly. We will test our hypothesis and attain the objective of this application by pursuing the following specific aims: (1) Synthesize molecules that can bind to androgen receptors and can form closely spaced 3MeA adducts, (2) Characterize the DNA-binding, DNA-methylating, and membrane permeability properties of the molecules, (3) Investigate the selectivity of the molecules for PCa cells overexpressing the AR and determine the role of AR in the observed toxicity, and (4) Examine the mechanism by which cellular toxicity is induced by the molecules in the PCa cells. The major outcome of our project will be the creation of molecules that can target PCa cells and form lethal DNA damage in them. These molecules will be used to probe AR- mediated delivery of toxins to DNA, investigate consequences of 3MeA and clustered adduct formation in PCa cells, and pave the way for future studies in animal models of PCa. This project will have a significant positive impact on the development of a new class of site-specific DNA-damaging agents that target specific cells and form DNA DSBs and lead to the development of targeted cancer chemotherapy agents with fewer side-effects. Another important outcome will be training of multiple undergraduate students, including underrepresented minorities, in organic synthesis, bioorganic and molecular biology techniques, tissue culture, and performing analytical and biological assays, thus providing them with advanced training in biomedical research and serving to enhance the biomedical research workforce.

抽象的我们研究的总体目标是制定可靠的策略，选择性地针对不同的目标癌细胞并在其中产生特定类型的 DNA 损伤，从而导致其有针对性的破坏。这些分子将帮助我们了解癌细胞中 DNA 损伤介导的生物过程，为疾病治疗的进步奠定基础。该 R15 提案的目标是开发靶向前列腺癌 (PCa) 细胞并在其中引起聚集性 DNA 损伤的分子，从而导致致命的 DNA 双链断裂 (DSB) 的形成。该项目解决了当前的关键问题 PCa 治疗需要开发具有更高选择性和效力的新化疗药物，但无毒副作用。我们的策略是合成与雄激素受体结合的分子 (AR) 在 PCa 细胞中过度表达，通过 AR 作用护送至这些细胞的细胞核，并且主要生成靠近的 N3-甲基腺嘌呤 (3MeA) DNA 加合物。细胞处理这些紧密排列的DNA损伤将导致DNA DSB的形成。该方法部分基于我们使用与雌激素结合的分子成功靶向破坏乳腺癌细胞受体（ER）在许多乳腺癌中过度表达。我们假设我们的策略将由于 DNA DSB 的致命性，导致对 PCa 细胞进行有针对性的、有效的破坏，并且因为 AR和ER属于同一核受体超家族，功能相似。我们将测试我们的假设并通过追求以下具体目标来实现本应用的目标：（1）合成可与雄激素受体结合并可形成紧密排列的 3MeA 加合物的分子，(2) 表征分子的 DNA 结合、DNA 甲基化和膜渗透性特性， (3) 研究分子对过表达AR的PCa细胞的选择性并确定其作用 AR 在观察到的毒性中的作用，以及 (4) 检查 AR 诱导细胞毒性的机制 PCa 细胞中的分子。我们项目的主要成果将是创造能够靶向 PCa 细胞并在其中形成致命的 DNA 损伤。这些分子将被用来探测 AR- 介导毒素向 DNA 的传递，研究 3MeA 和簇状加合物形成的后果 PCa细胞，为未来PCa动物模型的研究铺平道路。该项目将有一个对新型位点特异性 DNA 损伤剂的开发产生了重大积极影响靶向特定细胞并形成 DNA DSB，从而导致靶向癌症化疗的发展副作用较少的药物。另一个重要成果将是培养多名本科生有机合成、生物有机和分子生物学专业的学生，包括代表性不足的少数群体技术、组织培养以及进行分析和生物测定，从而为他们提供生物医学研究高级培训并服务于增强生物医学研究队伍。