Crosslinking-based targeted therapy for triple-negative breast cancer

摘要 HER 1和/或HER 3受体在大多数三阴性乳腺癌（TNBC）中过表达， 一种缺乏雌激素受体、孕酮和HER 2表达的乳腺癌亚型， 预后差，肺、脑转移率高。化疗是TNBC治疗的主要手段。 我们的长期目标是为这种侵袭性乳腺癌亚型开发更有效的治疗方法。在这 我们的目标是开发一种新的基于交联的靶向治疗。为此，我们将创建一个 一系列含有多个拷贝的HER 1和/或HER 3的多功能DNA-抗体-药物纳米颗粒 特异性抗体分子与DNA纳米结构共价偶联，后者与数十个小的 分子药物如THZ 1。在这些纳米颗粒的存在下，HER 1+和/或HER 3 + TNBC细胞将被诱导。 紧密结合在一起，从而在其初始阶段防止转移。在交联的TNBC细胞簇中， 与纳米颗粒可逆结合的小分子抗癌剂THZ 1将缓慢释放，导致 杀死交联的TNBC细胞。这些纳米颗粒可以在原代培养中结合至HER 1+和/或HER 3 + TNBC细胞。 肿瘤、转移部位和循环。在我们的初步研究中，将培养的HER 1 + TNBC细胞交联， 一起形成具有96%交联效率、99%迁移抑制和90%侵袭的细胞簇 通过DNA-4 ZHER 1-THZ 1纳米颗粒的抑制（药物与货物比为50）。 为了提高效率并克服对HER 1抑制剂的抗性，设计了一种新的DNA-2 ZHER 1 - 2 ZHER 3-THZ 1， 靶向双重HER 1和HER 3的纳米颗粒将在本研究中合成和测试。我们的设计和 假设将用以下具体目标进行检验：在具体目标1中，我们将制备DNA-2 ZHER 1 - 2 ZHER 3 - 2 ZHER-2 Z 用于靶向过表达HER 1和/或HER 3的TNBC的药物纳米颗粒。在具体目标2中，我们将评估 通过DNA-2 ZHER 1 - 2 ZHER 3-抑制HER 1-和HER 3-过表达的TNBC细胞生长和转移， 药物纳米颗粒体外研究。在具体目标3中，我们将研究所选DNA的分布和药代动力学， 2 ZHER 1 - 2 ZHER 3-药物纳米颗粒，并评估其对乳腺肿瘤生长的抑制和对肿瘤细胞增殖的抑制。 在TNBC异种移植物模型中的转移。 如果成功，这种创新的方法将为开发新的治疗方法开辟一条新的途径。 TNBC。本试验研究将为进一步研究其效用和机制奠定基础。 用于靶向治疗的新型抗癌剂。

Abstract The HER1 and/or HER3 receptors are overexpressed in most triple-negative breast cancers (TNBCs), a subtype of breast cancer that lacks estrogen receptor, progesterone, and HER2 expression and is associated with poor prognosis and a high lung and brain metastasis rate. Chemotherapy is the mainstay of TNBC treatment. Our long-term goal is to develop more effective therapy for this aggressive subtype of breast cancer. In this proposal, we aim to develop a novel crosslinking-based targeted therapy. For this purpose, we will create a series of multi-functional DNA-affibody-drug nanoparticles containing multiple copies of HER1 and/or HER3 specific affibody molecules covalently coupled to a DNA nanostructure, of which the latter binds to tens of small molecule drugs such as THZ1. In the presence of these nanoparticles, HER1+ and/or HER3+ TNBC cells will be tightly bound together, thus preventing metastasis in its initial stage. In the cross-linked TNBC cell clusters, the small molecule anticancer agent THZ1 reversibly bound to the nanoparticles will be released slowly, resulting in killing of crosslinked TNBC cells. These nanoparticles can bind to HER1+ and/or HER3+ TNBC cells in primary tumors, metastatic sites, and circulation. In our preliminary study, cultured HER1+ TNBC cells were crosslinked together to form cell clusters with a 96% crosslinking efficiency, 99% migration inhibition, and 90% invasion inhibition by a DNA-4ZHER1-THZ1 nanoparticle (drug-to-cargo ratio is 50). To enhance the efficiency and overcome the resistance to HER1 inhibitors, a novel DNA-2ZHER1-2ZHER3-THZ1 nanoparticle that targets dual HER1 and HER3 will be synthesized and tested in this study. Our design and hypothesis will be tested with the following specific aims: In specific aim 1, we will prepare a DNA-2ZHER1-2ZHER3- drug nanoparticle for targeting HER1- and/or HER3-overexpressing TNBC. In specific aim 2, we will evaluate the inhibition of HER1- and HER3-overexpressing TNBC cell growth and metastasis by the DNA-2ZHER1-2ZHER3- drug nanoparticle in vitro. In specific aim 3, we will study distribution and pharmacokinetics of selected DNA- 2ZHER1-2ZHER3-drug nanoparticle in healthy mice, and to evaluate its suppression of mammary tumor growth and metastasis in TNBC xenograft models. If successful, this innovative approach will open a new avenue for developing new therapy for treatment of TNBC. The pilot study will lay a foundation for further studies to investigate the utilities and mechanisms of the new class of anticancer agents for targeted therapy.