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

• 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
• 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

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.

摘要 我们研究的总体目标是开发一种可靠的策略，有选择地针对不同的 并在癌细胞中产生特定类型的DNA损伤，导致它们的靶向破坏。 这些分子将帮助我们了解DNA损伤介导的癌细胞和 为疾病治疗的进步奠定基础。此R15提案的目标是开发 靶向前列腺癌(PCA)细胞并导致其内部聚集的DNA损伤的分子导致 形成致命性DNA双链断裂(DSB)。该项目解决了当前的关键问题 前列腺癌治疗需要开发具有更高选择性和效力的新化疗药物，但 没有毒副作用。我们的策略是合成与雄激素受体结合的分子 (AR)在PCa细胞中过表达，通过AR作用被护送到这些细胞的细胞核，并且 主要在附近产生N3-甲基腺嘌呤(3MeA)DNA加合物。单元化处理 这些紧密相连的DNA损伤会导致DNA双链断裂的形成。这种方法部分地基于 在我们使用与雌激素结合的分子成功地靶向摧毁乳腺癌细胞之后 受体(ER)在许多乳腺癌中过度表达。我们假设我们的战略将 由于DNADSB的致命性，导致对PCA细胞的定向、有效的破坏 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申请的目标：(1) 合成能与雄激素受体结合并能形成紧密排列的3MeA加合物的分子，(2) 表征分子的DNA结合、DNA甲基化和膜通透性， (3)研究这些分子对高表达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)检查细胞毒性是由 前列腺癌细胞中的分子。我们项目的主要成果将是创造出能够 靶向前列腺癌细胞，并在其中形成致命的DNA损伤。这些分子将被用来探测AR- 介导毒素到DNA的传递，研究3MeA和簇状加合物形成的后果 为今后在前列腺癌动物模型中的研究奠定基础。这个项目将有一个 对开发一类新的位点特异性DNA损伤剂产生重大积极影响 靶向特定细胞并形成DNA双链构象并导致靶向肿瘤化疗的发展 副作用较少的药物。另一个重要成果将是培养多个本科生 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，并进行分析和生物分析，从而为他们提供 生物医学研究方面的高级培训，并为加强生物医学研究队伍服务。