Design and Application of Cationic Nanocarriers to Inhibit Breast Cancer Progression in Primary and Metastatic Sites

项目摘要 三阴性乳腺癌（TNBC）的特征在于缺乏雌激素/孕激素受体， 人表皮生长因子受体2（HER 2）表达及其高复发率， 转移化疗仍然是乳腺癌治疗的主要支柱之一，特别是对于 三阴性乳腺癌虽然化疗有利于杀死恶性肿瘤细胞，但它会导致 到损伤相关分子模式（DAMP）的释放。DAMP是一个促成因素， 癌症相关炎症，可通过几种机制增强未来的转移性扩散 例如转移部位的肿瘤微环境（TMEM）的发展。这些DAMP包括 核酸、细胞因子和蛋白质如HMGB 1。聚酰胺胺（PAMAM）是一种可生物降解的， 水溶性树枝状聚合物具有根据其不同的电荷和尺寸的能力， 末端分支和分支度（即世代）。胺封端的PAMAM是 带正电荷（即阳离子）并且可以结合DNA和RNA。在这个树枝状聚合物的基础上， 合成的具有降低的毒性的改性阳离子PAMAM-G 3衍生物 并且可以将化学药物包封为纳米颗粒并保持核酸结合性质。我们 初步试验表明，这些材料可以与游离DNA和RNA结合， 这是用化疗如阿霉素和紫杉醇治疗三阴性乳腺癌细胞的结果。 在这项研究计划中，我们的目标是探索我们的材料可以与哪些其他化疗诱导的DAMP结合 和压制。将使用体外和体内模型研究材料的抗转移作用 以及患者血清样本。小鼠转移性乳腺癌模型将作为以下研究的基础： 评估传统化疗输送与化疗输送相比的效果， PAMAM-G3纳米颗粒在原发性肿瘤生长、转移程度和炎性方面的作用 小鼠血清中的材料。总之，我们建议追求以下具体目标：（1）表征 从化疗处理的TNBC细胞释放的损伤相关分子模式（DAMP）; (2)测定PAMAM-G3清除聚合物和纳米颗粒对人结肠癌的治疗功效。 由化疗诱导的DAMP引起的免疫系统激活和侵袭潜力;和（3） 了解PAMAM-G3介导DAMP清除的机制。这个实验 该提案将有助于对化疗如何影响循环前体的新知识， 转移性DAMP。此外，一种新的双重化疗递送和DAMP清除方法， 将研究修饰的PAMAM-G3纳米颗粒在减少原发性肿瘤和转移性肿瘤中的效用。 负担在哥伦比亚大学完成本计划书将为申请人提供癌症培训 医学生物学和工程学，为成为一名独立研究者做准备。

PROJECT SUMMARY Triple-negative breast cancer (TNBC) is characterized by the lack of estrogen/progesterone receptors and human epidermal growth factor receptor 2 (HER2) expression as well as its high rates of recurrence and metastasis. Chemotherapy persists as one of the mainstays of breast cancer treatment, particularly for triple-negative breast cancer. While chemotherapy is beneficial for killing the malignant tumor cells, it leads to the release of damage-associated molecular patterns (DAMPs). DAMPs are a contributing factor to cancer-related inflammation which can potentiate future metastatic spread through several mechanisms such as the development of tumor microenvironments of metastasis (TMEM) sites. These DAMPs include nucleic acids, cytokines, and proteins such as HMGB1. Polyamidoamine (PAMAM) is a biodegradable, water-soluble dendrimer polymer with the ability to possess different charges and sizes depending on its terminal branches and degree of branching (i.e. generation), respectively. Amine-terminated PAMAM is positively charged (i.e. cationic) and can bind DNA and RNA. Building on this dendrimer, we have synthesized modified cationic PAMAM-generation 3 (PAMAM-G3) derivatives that have decreased toxicity and can encapsulate chemodrugs as nanoparticles and maintain the nucleic acid-binding property. Our preliminary tests have shown that these materials can bind to both cell-free DNA and RNA released as a result of treating triple-negative breast cancer cells with chemotherapy such as doxorubicin and paclitaxel. In this research plan we aim to explore what other chemotherapy-induced DAMPs our materials can bind to and suppress. The anti-metastatic effects of the materials will be studied using in-vitro and in-vivo models as well as patient serum samples. A murine metastatic breast cancer model will serve as the basis for assessing the effects of traditional chemotherapy delivery compared with chemotherapy delivery using PAMAM-G3 nanoparticles with respect to primary tumor growth, degree of metastasis, and inflammatory materials in mouse serum. In summary, we propose to pursue the specific aims of (1) Characterize damage-associated molecular patterns (DAMPs) released from chemotherapy-treated TNBC cells; (2) Determine the therapeutic efficacy of PAMAM-G3 scavenging polymers and nanoparticles on immune system activation and invasive-potential caused by chemotherapy-induced DAMPs; and (3) Understand the mechanisms behind PAMAM-G3 mediated DAMP scavenging. The experiments in this proposal will contribute new knowledge on how chemotherapy influences the profile of circulating pro- metastatic DAMPs. In addition, a novel method of dual chemotherapy delivery and DAMP scavenging via modified PAMAM-G3 nanoparticles will be studied for its utility in reducing primary tumor and metastatic burden. Completion of this proposal at Columbia University will provide the applicant with training in cancer biology and engineering in medicine in preparation to becoming an independent investigator.