Prevention of post-therapy breast cancer metastasis

尽管在标准治疗方面取得了进展，但30%-40%的早期乳腺癌患者会发展为 治疗转移。乳腺癌干细胞(BCSC)可引起转移并对标准耐药 化疗和放射治疗。此外，放射治疗(IR)或化疗(化疗)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 耐药BCSC和治疗诱导BCSC的阻断形成。BCSC/iBCSC Express均已提升 乙醛脱氢酶(ALDH)水平。双硫兰(DSF)是FDA批准的ALDH抑制剂，用于 酒精中毒的治疗。它对乳腺癌细胞的毒性通过与必需的痕量结合而增强 元素铜形成DSF/铜络合物。DSF/Cu是一种有效的 蛋白酶体抑制物 导致 抑制关键转录因子NF-κB，它与癌症和放射、化疗耐药和 与最新的流行病学报告一致，表明DSF显著减少 患者死于癌症的风险，我们发现DSF和IR有效地靶向BSCs/iBCSC，并显著 在侵袭性小鼠乳腺肿瘤4T1模型中阻止肺转移，而IR单独治疗 效果不佳。我们还发现，DSF/Cu在体外和体内都能通过向下阻断IR或化疗诱导的BCSC。 降低促生存κ活性对核因子-乳酸脱氢酶B干性基因通路和靶向bcsc的调节 诱导内质网(ER)应激-免疫细胞死亡(ICD)。此外，DSF和IR 在4T1小鼠模型中诱导了对原发肿瘤和肺转移的强大免疫反应。 ICD 细胞干性。 这些 研究结果为我们的假设提供了理论基础，DSF/Cu导致耐药的BCSC和 阻断IR或化疗诱导的BCSC的形成，在以下情况下有效地防止乳腺癌转移 结合最常用的标准治疗，即手术、IR和化疗。 既然铜是肿瘤 促进和发现乳腺癌患者肿瘤和血清中水平升高，量身定做使用外源性 体内含DSF的铜将以所有靶点的肿瘤铜水平为基础。 在Aim1中，我们将评估治疗 IR和/或化疗联合DSF/Cu预防乳腺癌转移的疗效观察 佐剂环境下的异种移植(PDX)和MMTV-PYMT转基因乳腺肿瘤。在AIM2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 新佐剂环境中的肿瘤。在Aim3中，我们将 系统地分析了DSF/Cu的作用机制 诱导ICD和调节IR诱导的免疫应答靶向BCSC 。在此结束时 在这项研究中，我们将有：i)评估DSF/铜作为一种在外科环境中靶向BCSC/iBCSC的新型药物， 化疗和/或IR治疗；二) DSF/Cu和IR诱导强健免疫的机制研究 对乳腺癌转移的反应；以及iii)形成了设计临床之前的基础 研究计划，以测试这一新的治疗方法，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.