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

摘要 基于纳米颗粒的技术，包括治疗诊断纳米医学，是生物医学的快速发展领域。 为癌症治疗提供前所未有的潜力。该应用程序旨在将此 潜在的翻译水平，并研究我们的新的成像能力纳米药物的治疗应用 转移性乳腺癌在一个大的动物模型。乳腺癌转移是导致死亡的主要原因 在乳腺癌患者中，并且由于该原因，本申请的特定目的响应于未满足的 临床需要这一策略的提出是基于我们的发现，即转移性细胞的存活至关重要， 依赖于microRNA-10 b（miR-10 b）的高表达，miR-10 b负责转移性细胞 生存能力。随着这一发现，通过抑制miR-10 b在转移部位杀死这些细胞成为可能。这 使用由与葡聚糖包被的磁性载体缀合的抗miR-10 b寡核苷酸组成的纳米药物来实现 纳米颗粒（称为MN-抗-miR 10 b）。这些纳米颗粒作为递送载体用于Escheromir。还有， 这些纳米颗粒的磁性允许使用磁性纳米颗粒监测它们在体内的递送 磁共振成像（MRI），这是该治疗方法的临床实施的附加值。在 在小鼠的临床前研究中，我们显示MN-抗-miR-10 b递送至淋巴中已建立的转移灶， 淋巴结，肺，骨骼和大脑在伴侣猫中的初步研究表明，将纳米药物递送到 静脉注射后转移病灶。人MDA-MB-231鼠模型中的治疗研究 TNBC显示MN-抗miR 10 b递送至具有已确定转移的淋巴结导致miR 10 b的抑制。 通过停止肿瘤细胞增殖和触发凋亡来转移生长。当与低剂量的 化疗后，MN-抗miR 10 b引起淋巴结转移完全消除，没有证据表明 在四周治疗后出现全身毒性。接下来，我们展示了纳米药物与 低剂量化疗用于治疗已确定的肺转移，对应于人类IV期 免疫活性鼠模型（4 T1）中的疾病。我们发现每周六次的治疗足以引起 预先存在的肺转移完全消退，没有进一步扩散到其他器官， 剩下的动物。 展望临床翻译我们的研究，在这个应用程序中，我们建议测试我们的纳米药物策略 在患有自发性转移性乳腺癌的大型动物（伴侣猫）中。这里提出的研究将有助于 这是迈向首次人体试验的必要一步，因为它们将证明纳米药物的成功递送， 大型动物，其在解剖学和生理学上不同于迄今为止测试的鼠模型。 此外，这些研究将为我们提供关于大型动物PK/PD参数的额外信息 模型最后，在自发模型中的治疗研究将模仿人类中的那些， 转移在时间和位置上很大程度上是不可预测的。我们希望这些研究将作为 本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.