Prevention of post-therapy breast cancer metastasis

尽管标准疗法取得了进展，但30-40%的早期乳腺癌患者将发展为晚期乳腺癌。 治疗转移。乳腺癌干细胞（BCSC）会引起转移，并对标准治疗有抗性。 化疗和放疗。此外，放射（IR）或化疗（chemo）可以重新编程 将非干细胞乳腺癌细胞转化为BCSC，即iBCSC。因此，有效的癌症治疗必须消除 治疗抗性BCSC和阻断治疗诱导的BCSC的形成。BCSC/iBCSC表达升高 乙醛脱氢酶（ALDH）。双硫仑（DSF）是FDA批准的ALDH抑制剂，用于治疗糖尿病。 治疗酒精中毒。其对乳腺癌细胞的毒性因与必需微量元素的结合而增强 元素铜（Cu）形成DSF/Cu络合物。DSF/Cu是一种有效的 蛋白酶体抑制剂 导致 抑制关键转录因子NF-κB，其与癌症和放射性、化学抗性相关， 与最近的流行病学报告一致，表明DSF显著降低 我们发现，DSF和IR有效靶向BSCS/iBCSC，并显著降低了患者的癌症死亡风险。 预防侵袭性小鼠乳腺肿瘤4 T1模型的肺转移，而单独IR 无效。我们还发现，DSF/Cu可以阻断体外和体内IR或化学诱导的BCSC，通过下调- 通过降低促存活ALDH活性调节NF-κ B-干性基因通路和靶向BCSC 和诱导内质网（ER）应激-免疫细胞死亡（ICD）。此外，DSF和IR 在4 T1小鼠模型中诱导针对原发性肿瘤和肺转移的稳健免疫应答。 ICD 干细胞性 这些 这些发现为我们的假设提供了理论基础，即DSF/Cu诱导了治疗抵抗性BCSC， 阻断IR或化疗诱导的BCSC的形成，有效预防乳腺癌转移， 结合最常用的标准治疗，即，手术IR和化疗 既然Cu是肿瘤 促进并发现在乳腺癌患者的肿瘤和血清中水平升高， 在所有目的中，体内DSF的Cu将基于肿瘤Cu水平。 在目标1中，我们将评估治疗性 DSF/Cu联合IR和/或化疗预防乳腺癌转移疗效观察 异种移植物（PDX）和MMTV-PyMT转基因乳腺肿瘤在佐剂环境中的生长。在目标2中，我们将评估 化疗联合DSF/Cu预防PDX和MMTV-PyMT转移的疗效观察 新辅助治疗中的肿瘤。在AIM 3中，我们将 系统分析了DSF/Cu复合材料的作用机理， 通过诱导ICD和调节IR诱导的免疫应答靶向BCSC .在这一结论 i）评估DSF/Cu作为手术背景下靶向BCSC/ iBCSC的新型药物， 化学和/或IR治疗; ii） 阐明了DSF/Cu和IR诱导强大免疫的机制， 对乳腺癌转移的反应;和iii）形成了设计临床治疗方案的临床前基础。 研究计划，以测试这种新的疗法，以防止治疗后乳腺癌转移。

Despite advances in standard therapies,30-40% of patients with early-stage breast cancer will develop post- therapy metastases. Breast cancer stem cells (BCSC) give rise to metastases and are resistant to standard chemotherapy and radiation therapy. Moreover, radiation (IR) or chemotherapy (chemo) can reprogram nonstem breast cancer cells into BCSC, namely iBCSC. Thus, effective cancer treatment must eliminate therapy-resistant BCSC and block formation of therapy-induced BCSC. Both BCSC/iBCSC express elevated levels of aldehyde dehydrogenase (ALDH). Disulfiram (DSF) is an FDA-approved inhibitor of ALDH for treatment of alcoholism. Its toxicity to breast cancer cells is enhanced by the binding of the essential trace element copper (Cu) to form DSF/Cu complexes. DSF/Cu is an effective proteasome inhibitor resulting in inhibition of the key transcriptional factor NF-κB, which is linked to cancer and radio-, chemo-resistance and Consistent with the most recent epidemiological report indicating that DSF significantly reduced patient risk of death from cancer, we found that DSF and IR effectively targets BSCS/iBCSC and significantly prevented lung metastasis in the aggressive mouse mammary tumor 4T1 model, while IR alone was ineffective. We also found that DSF/Cu can block in vitro and in vivo IR- or chemo-induced BCSC via down- regulation of the NF-κB-stemness gene pathway and target BCSC by reduction of pro-survival ALDH activity and induction of endoplasmic reticulum (ER) stress- immunological cell death (ICD). Moreover, DSF and IR induced a robust immune response against primary tumor and lung metastasis in 4T1 mouse models. ICD cell stemness. These findings provide the rationale for our hypothesis that DSF/Cu, which induces of therapy-resistant BCSC and blocks formation of IR- or chemo- induced BCSC, is effective in preventing breast cancer metastasis when combined with the most frequently used standard treatment, i.e., surgery, IR and chemo. Since Cu is tumor promoting and found at elevated levels in tumors and sera of breast cancer patents, tailored use of exogenous Cu with DSF in vivo will be based on tumor Cu level in all Aims. In Aim1, we will assess the therapeutic efficacy of IR and/or chemo combined with DSF/Cu on preventing metastasis of patient breast cancer-derived xenograft (PDX) and MMTV-PyMT transgenic mammary tumors in an adjuvant setting. In Aim2, we will assess the therapeutic efficacy of chemo combined with DSF/Cu on preventing metastasis of PDX and MMTV-PyMT tumors in a neoadjuvant setting. In Aim3, we will analyze the mechanisms by which DSF/Cu systemically targets BCSC via induction of ICD and modulation of IR-induced immune response . At the conclusion of this study, we will have: i) evaluated DSF/Cu as a novel agent targeting BCSC/ iBCSC in the context of surgery, chemo-and/or IR treatment; ii) elucidated mechanisms by which DSF/Cu and IR induce a robust immune response against breast cancer metastasis; and iii) formed the preclinical foundation for designing a clinical research program to test this novel therapy to prevent post-therapy breast cancer metastasis.