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

摘要 我们研究的总体目标是开发一种可靠的策略， 癌细胞并在其中产生特定类型的DNA损伤，导致其靶向破坏。 这些分子将帮助我们了解癌细胞中DNA损伤介导的生物学过程， 为疾病治疗的进步奠定基础。本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， 形成致命的DNA双链断裂（DSB）。该项目解决了当前的关键问题， 需要在PCa治疗中开发具有更高选择性和效力的新化疗剂， 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细胞中过表达的，通过AR作用被护送到这些细胞的细胞核， 主要产生N3-甲基腺嘌呤（3 MeA）DNA加合物。细胞加工 这些紧密间隔的DNA损伤将导致DNA DSB的形成。这种方法部分基于 在我们成功地用与雌激素结合的分子靶向破坏乳腺癌细胞后， 受体（ER）在许多乳腺癌中过表达。我们假设我们的策略 由于DNA DSB的致命性， AR和ER属于相同的核受体超家族并且功能相似。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的：（1） 合成可与雄激素受体结合并可形成紧密间隔的3 MeA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 观察到的毒性中的AR，和（4）检查细胞毒性由AR诱导的机制。 PCa细胞中的分子。我们项目的主要成果将是创造分子， 靶向PCa细胞并在其中形成致命的DNA损伤。这些分子将用于探测AR- 介导的毒素传递到DNA，研究3 MeA和成簇加合物形成的后果， PCa细胞，并为未来的研究在PCa的动物模型。该项目将有一个 对一类新的位点特异性DNA损伤剂的开发产生了显著的积极影响， 靶向特异性细胞并形成DNA双链断裂，导致靶向癌症化疗的发展 药物副作用少。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，并进行分析和生物测定，从而为他们提供 生物医学研究的高级培训和服务，以提高生物医学研究的劳动力。

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.