anti-miR-10b Nanodrug for Treatment of Breast Cancer Metastasis: Study in Companion Animals

摘要 以纳米颗粒为基础的技术是生物医学中发展最快的领域 研究为癌症治疗提供了前所未有的潜力。此应用程序旨在实现以下功能 潜在的翻译水平，并研究我们的新型可成像纳米片在治疗中的应用 在一个大型动物模型中发现了转移性乳腺癌。乳腺癌转移是导致死亡的主要原因 在乳腺癌患者中，出于这个原因，这项应用的特定目标回应了一个未实现的 临床需要。这一提出的策略是基于我们的发现，转移细胞的生存至关重要 依赖于microRNA-10b(miR-10b)的高表达以及miR-10b对转移细胞的作用 生存能力。有了这一发现，就有可能通过抑制miR-10b在转移部位杀死这些细胞。这 是使用由抗miR-10b对映体与葡聚糖包被的磁性结合而成的纳米地毯来实现的 纳米粒(称为MN-抗miR10b)。这些纳米颗粒充当了异形虫的运输工具。另外， 这些纳米颗粒的磁性允许使用磁学技术监测它们在体内的传递 磁共振成像(MRI)，这是临床实施这一治疗方法的附加值。在……里面 在小鼠的临床前研究中，我们显示了MN-抗miR-10b对已确定的淋巴转移瘤的输送 结节、肺、骨和脑。在伴侣猫身上的初步研究表明，纳米地毯被送到了 静脉注射后转移灶。人MDA-MB-231小鼠模型的治疗研究 TNBC显示，将MN-抗miR10b转移到已确定转移的淋巴结可导致抑制 通过阻止肿瘤细胞增殖和引发细胞凋亡而发生转移生长。当与低剂量的 化疗后，MN-抗miR10b导致的淋巴转移完全消除，没有证据表明 仅在每周四次治疗后出现全身毒性。接下来，我们展示了纳米地毯的组合用途 小剂量化疗治疗已确诊的IV期人肺转移瘤 免疫活性小鼠模型中的疾病(4T1)。我们发现每周六次的治疗足以引起 原存在的肺转移完全消退，不会进一步扩散到其他器官 剩下的动物。 展望我们研究的临床翻译，在这个应用中，我们建议测试我们的纳米芯片策略 在患有自发性转移性乳腺癌的大型动物(伴猫)中。这里提出的研究将有助于 作为第一次人体试验的必要步骤，因为他们将证明成功地在 大型动物，它们在解剖学和生理学上与迄今测试的小鼠模型截然不同。 此外，这些研究将为我们提供更多关于大型动物PK/PD参数的信息 模特。最后，在自发模型中的治疗性研究将模仿人类的治疗研究，其中 转移在时间和位置上很大程度上是不可预测的。我们预期这些研究将会成为 该项目临床前阶段的最后一步，下一步将进入IND前阶段。

ABSTRACT Nanoparticle-based technologies including theranostic nanomedicine are fast developing areas of biomedical research offering unprecedented potential for cancer treatment. This application is designed to bring this potential to translational level and investigate the application of our novel image-capable nanodrug for treatment of metastatic breast cancer in a large animal model. Breast cancer metastasis is the main cause of mortality among breast cancer patients, and for that reason the specific aims of this application respond to an unmet clinical need. This proposed strategy is based on our discovery that the survival of metastatic cells crucially depends on the high expression of microRNA-10b (miR-10b) and that miR-10b is responsible for metastatic cell viability. With this discovery it became possible to kill these cells at metastatic sites by inhibiting miR-10b. This is achieved using a nanodrug composed of anti-miR-10b antagomirs conjugated to dextran-coated magnetic nanoparticles (termed MN-anti-miR10b). These nanoparticles serve as delivery vehicles for the antagomirs. Also, the magnetic properties of these nanoparticles allow for monitoring of their delivery in vivo using magnetic resonance imaging (MRI), which is an added value for clinical implementation of this therapeutic approach. In pre-clinical studies in mice, we showed delivery of MN-anti-miR-10b to established metastases in the lymph nodes, lungs, bone and brain. Pilot studies in companion cats showed the delivery of the nanodrug to the metastatic lesions after intravenous injection. Therapeutic studies in the murine model of human MDA-MB-231 TNBC showed that delivery of MN-anti-miR10b to lymph nodes with established metastases resulted in arrest of metastatic growth by halting tumor cell proliferation and triggering apoptosis. When combined with a low-dose chemotherapy, MN-anti-miR10b caused complete elimination of lymph node metastases with no evidence of systemic toxicity after just four weekly treatments. Next, we showed the utility of the nanodrug in combination with low-dose chemotherapy for treatment of established lung metastases corresponding to Stage IV of human disease in immunocompetent murine model (4T1). We found that six weekly treatments were sufficient to cause complete regression of pre-existing lung metastases with no further dissemination to other organs in the remaining animals. With an outlook to clinical translation of our studies, in this application we propose to test our nanodrug strategy in large animals with spontaneous metastatic breast cancer (companion cats). Studies proposed here will serve as a necessary step towards first-in-human trials, because they will prove successful delivery of the nanodrug in large animals, which are anatomically and physiologically distinct from the murine models tested to date. Furthermore, these studies will provide us with additional information on PK/PD parameters in a large animal model. Finally, therapeutic studies in spontaneous models will mimic those in humans where development of metastases is largely unpredicted in terms of time and location. We expect that these studies will serve as the final step in the pre-clinical stage of this project, which will next move to the pre-IND stage.