Multifunctional Nanotherapeutics for Cancer Treatment and Imaging

描述（由申请人提供）： 原发性卵巢癌，特别是腹水中生长的转移性细胞的化疗治疗的成功受到癌细胞的内在和获得性抗性以及化疗的不良副作用的限制。基于我们先前研究的结果和文献数据，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）通过靶向在这种抗性中起关键作用的蛋白质的siRNA抑制癌细胞抗性和（3）靶向药物，特异性针对原发肿瘤和转移灶中卵巢癌细胞的成像剂和siRNA。只有将几种抗癌药物与其他活性成分组合递送至卵巢肿瘤细胞，才能实现这样的目的，所述其他活性成分执行不同的特定功能，以增强细胞摄取和主要药物在癌细胞中的效率，限制不良副作用，并防止现有耐药性的发展和/或抑制。在拟议的研究中，我们计划将纳米技术方法应用于这种多组分多功能纳米治疗药物的开发和评估。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），这将显着提高原发性卵巢癌和腹膜内转移化疗的疗效，同时最大限度地减少对健康器官的副作用。羟基封端的PAMAM-OH和内部季铵化和表面乙酰化的聚（酰胺基胺）树枝状聚合物（QPAMAM-NHAc）将分别用作递送抗癌药物和siRNA的纳米载体。此外，每个NDDS将含有肿瘤特异性靶向部分（肽）和一种活性组分（抗癌药物或siRNA或荧光染料）。紫杉醇和顺铂/卡铂将作为抗癌药物-细胞死亡诱导剂进行评价。将研究靶向MDR 1和CD 44 mRNA的siRNA作为泵抗性的抑制剂。靶向BCL 2 mRNA的siRNA将作为非泵抗性的抑制剂进行研究。促黄体生成激素释放激素（LHRH）肽将用作卵巢癌特异性靶向部分。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的细胞将用于在裸鼠中建立异位皮下和原位腹膜内模型。将比较NDDS的静脉全身和腹膜内局部给药。拟议研究的结果将用于设计用于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纳米技术为基础的方法，有效治疗卵巢癌的同时抑制细胞耐药的siRNA靶向MDR 1，CD 44和BCL 2 mRNA和细胞死亡诱导两种抗癌药物（紫杉醇和顺铂）具有不同的作用机制。拟议研究的目的是开发一种基于多功能纳米载体的药物递送系统的新型混合物（“鸡尾酒”），其将显著提高原发性卵巢癌和腹膜内转移的化疗的功效，同时最大限度地减少治疗对健康器官的副作用。

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.