Multifunctional Nanotherapeutics for Cancer Treatment and Imaging

描述(由申请人提供)： 原发卵巢癌，尤其是腹水中生长的转移细胞的化疗成功受到癌细胞固有和获得性耐药以及化疗副作用的限制。根据我们以前的研究结果和文献数据，我们假设，通过(1)至少两种不同的抗癌药物诱导癌细胞死亡，(2)针对在耐药中起关键作用的蛋白质的siRNA抑制癌细胞耐药，(3)针对原发肿瘤和转移瘤中的卵巢癌细胞的靶向药物、显像剂和siRNA，可以显著提高耐药卵巢癌和转移瘤的治疗和成像的有效性。只有将几种抗癌药物与发挥不同特定功能的其他活性成分结合在一起，才能实现这一目标，这些活性成分可增强细胞对主要药物的摄取和效率，特别是在癌细胞中，限制不良副作用，并防止和/或抑制现有耐药性的发展。在拟议的研究中，我们计划将纳米技术方法应用于此类多组分多功能纳米疗法的开发和评估。这项拟议研究的长期目标是验证这一假设，并开发一种新型多功能纳米技术药物递送系统(NDDS)的混合物(鸡尾酒)，它将显著提高原发卵巢癌和腹膜转移癌的化疗效率，同时将对健康器官的副作用降至最低。一种端羟基的PAMAM-OH和内部季胺化和表面乙酰化的聚(氨基胺)树状大分子(QPAMAM-NaAC)将分别作为纳米载体输送抗癌药物和siRNA。此外，每个NDDS将包含一个肿瘤特异性靶向部分(肽)和一个活性成分(抗癌药物或siRNA或荧光染料)。紫杉醇和顺铂/卡铂将被评估为抗癌药物-细胞死亡诱导剂。针对MDR1和CD44 mRNA的siRNA将作为泵阻力的抑制因子进行研究。靶向bcl2mRNA的siRNA将作为非泵阻力的抑制因子进行研究。促黄体生成素释放激素(LHRH)多肽将被用作卵巢癌特异性靶向部分。已建立的人多药耐药卵巢癌细胞系以及从晚期多药耐药卵巢癌患者的原发肿瘤和恶性腹水中分离出的细胞将被用于建立裸鼠异位皮下和原位腹膜内模型。我们将比较静脉注射、全身注射和腹膜内局部注射NDDS。拟议的研究结果将用于设计治疗不同癌症的新型多功能纳米技术方法。 公共卫生相关性： 我们提出了一种新的基于纳米技术的有效治疗卵巢癌的方法，通过靶向mdr1、CD44和bcl2基因的siRNA抑制细胞耐药，并用两种不同作用机制的抗癌药物(紫杉醇和顺铂)诱导细胞死亡。这项研究的目的是开发一种新型的多功能纳米载体给药系统的混合物(“鸡尾酒”)，它将显著提高原发性卵巢癌和腹膜转移癌的化疗效果，同时将治疗对健康器官的副作用降至最低。

DESCRIPTION (provided by applicant): The success of chemotherapeutic treatment of primary ovarian cancer, especially metastatic cells growing in ascitic fluid is limited by intrinsic and acquired resistance of cancer cells and adverse side effects of chemotherapy. Based on the results of our previous study and literature data, we hypothesized, that substantial enhancement in the effectiveness of treatment and imaging of drug resistant ovarian cancer and metastases can be achieved by the (1) induction of cancer cell death by at least two different anticancer drugs, (2) suppression of cancer cell resistance by siRNA targeted to proteins that play key roles in such resistance and (3) targeting drugs, imaging agents and siRNA specifically to ovarian cancer cells in primary tumor and metastases. Such an objective can only be achieved if several anticancer drugs are delivered to the ovarian tumor cells in combination with other active components that perform different specific functions for enhancing cellular uptake and efficiency of the main drugs specifically in cancer cells, limiting adverse side effects, and preventing the development and/or suppression of the existing drug resistance. In the proposed study, we plan to apply nanotechnology approaches to the development and evaluation of such multicomponent multifunctional nanotherapeutics. The long-term objective of the proposed research is to verify the hypothesis and develop a mixture (cocktail) of novel multifunctional Nanotechnology-based Drug Delivery Systems (NDDS) that will significantly increase the efficacy of the chemotherapy of primary ovarian cancer and intraperitoneal metastases while minimizing side effects on healthy organs. A hydroxyl terminated PAMAM-OH and internally quaternized and surface-acetylated Poly(amido amine) dendrimer (QPAMAM-NHAc) will be used as a nanocarrier to deliver anticancer drugs and siRNA, respectively. In addition, each NDDS will contain a tumor-specific targeting moiety (peptide) and one active component (anticancer drug or siRNA or fluorescent dye). Paclitaxel and cisplatin/carboplatin will be evaluated as anticancer drugs - cell death inducers. siRNA targeted to MDR1 and CD44 mRNA will be investigated as suppressors of pump resistance. siRNA targeted to BCL2 mRNA will be studied as a suppressor of nonpump resistance. Luteinizing Hormone-Releasing Hormone (LHRH) peptide will be used as ovarian cancer-specific targeting moiety. Established human multidrug resistant ovarian cancer cell lines as well as cells isolated from primary tumor and malignant ascites from patients with advanced multidrug resistant ovarian carcinoma will be used to create ectopic subcutaneous and orthotopic intraperitoneal models in nude mice. Intravenous systemic and intraperitoneal local administrations of NDDS will be compared. The results of the proposed research will be used to design novel multifunctional nanotechnology approaches for the treatment of different cancers. PUBLIC HEALTH RELEVANCE: We are proposing a novel nanotechnology-based approach for the effective treatment of ovarian cancer by simultaneous suppression of cellular resistance by siRNA targeted to MDR1, CD44 and BCL2 mRNA and cell death induction by two anticancer drugs (paclitaxel and cisplatin) with different mechanisms of action. The objective of the proposed research is to develop a novel mixture ("cocktail") of multifunctional nanocarrier- based drug delivery systems that will significantly increase the efficacy of the chemotherapy of primary ovarian cancer and intraperitoneal metastases while minimizing side effects of treatment on healthy organs.