Nanoscale Coordination Polymers Co-deliver Chemotherapeutics and siRNAs for Efficacious Treatment of Resistant Ovarian Cancer

项目摘要： 卵巢癌（OCa）是美国女性癌症死亡的第五大原因。本质 耐药和复发性卵巢癌是用现有疗法不能治愈的晚期疾病。 考虑到肿瘤的分子和遗传异质性，我们假设有效的OCa治疗 可以通过确定在个体OCa患者的肿瘤中发现的遗传异常来开发， 设计能够克服患者特异性多药耐药（MDR）的个性化治疗。我们建议 开发稳定的纳米级配位聚合物（NCP），用于一线和二线OCa的共同输送 化疗剂（顺铂或顺铂加吉西他滨）和靶向MDR基因的siRNA混合物。一个独特 内体逃逸机制将得到阐明，并有望普遍适用于合理的设计 用于生物制剂在体内的有效递送的纳米载体。该项目不仅将提供新的基本见解 纳米医学研究，但也有很大的希望，为临床翻译的个性化治疗， 抗OCa。 目的1：构建NCP/siRNA并评价其对卵巢癌耐药细胞系的体外作用。两个NCP/siRNA 将开发在核中具有化疗剂并且在壳中具有靶向MDR的siRNA的制剂， 表征了将在耐药OCa细胞中评价体外基因沉默和细胞毒性。 目的2：评价NCP/siRNA在耐药OCa原位小鼠肿瘤模型中的抗癌效果。的 将在小鼠模型中评估NCP/siRNA的一般毒性和抗癌功效。 目的3：通过将Her 2/neu抗体片段整合到NCP/siRNA外壳中来主动靶向NCP/siRNA。 小蛋白质Her 2/neu抗体片段将与脂质缀合并掺入脂质的外壳中。 将在Her 2/neu高表达和Her 2/neu低表达中测试NCP/siRNA和所得纳米颗粒。 表达耐药OCa肿瘤模型以评估药代动力学、肿瘤沉积和功效的变化。 目的4：评价NCP/siRNA和NCP/siRNA/H2 A在患者来源的异种移植物（PDX）中的抗癌功效 耐OCa的小鼠模型。将在OCa肿瘤PDX中分析MDR相关基因的表达 马约诊所收集的样本。将设计需要化疗药物的个性化治疗 以及靶向MDR基因的siRNA的选择。确认的铂耐药肿瘤细胞将用于评估 NCP/siRNA在原位PDX OCa小鼠模型中的抗癌功效。Her-2/neu受体肿瘤 阳性和阴性患者将通过NCP/siRNA/H2 A进一步评估抗癌功效。 通过这些目标，我们寻求建立一个新的范式，为耐药OCa的治疗。NCP/siRNA 可为OCa患者提供个性化治疗，并在耐药患者中实现大大增强的抗癌疗效， OCa。作为宫颈癌和其他癌症的护理标准，基于顺铂的NCP/siRNA可以具有广泛的应用前景。 对治疗其他耐药癌症的影响。

Project Summary: Ovarian cancer (OCa) is the fifth leading cause of cancer death for women in the United States. Intrinsically resistant and recurring ovarian cancers are a terminal disease that cannot be cured with existing therapeutics. Considering the molecular and genetic heterogeneity of tumors, we hypothesize that efficacious OCa therapy can be developed by determining the genetic abnormalities found in tumors of individual OCa patients and designing personalized therapy that can overcome patient-specific multi-drug resistance (MDR). We propose to develop robust nanoscale coordination polymers (NCPs) for the co-delivery of front-line and second-line OCa chemotherapeutics (cisplatin or cisplatin plus gemcitabine) and siRNA cocktails targeting MDR genes. A unique endosomal escape mechanism will be elucidated and is expected to be generally applicable to the rational design of nanocarriers for efficient delivery of biologics in vivo. This project will not only provide new fundamental insights nanomedicine research, but also holds great promise for clinical translation for the personalized treatment of resistant OCa. Aim 1: Develop NCP/siRNAs and evaluate their in vitro effects on resistant OCa cell lines. Two NCP/siRNAs formulations with chemotherapeutics in the core and siRNAs targeting MDR in the shell will be developed and characterized. The in vitro gene silencing and cytotoxicity will be evaluated in resistant OCa cells. Aim 2: Evaluate the anticancer efficacy of NCP/siRNAs in orthotopic mouse tumor models of resistant OCa. The general toxicity and anticancer efficacy of NCP/siRNAs will be assessed in mouse models. Aim 3: Actively target NCP/siRNAs by incorporating Her2/neu antibody fragments into the NCP/siRNAs shell. Small protein Her2/neu antibody fragments will be conjugated to a lipid and incorporated into the outer shell of NCP/siRNAs and the resulting nanoparticles will be tested in Her2/neu high expressing and Her2/neu low expressing resistant OCa tumor models to evaluate changes in pharmacokinetics, tumor deposition, and efficacy. Aim 4: Evaluate the anticancer efficacy of NCP/siRNAs and NCP/siRNAs/H2A in patient-derived xenograft (PDX) mouse models of resistant OCa. The expression of MDR-associated genes will be analyzed in OCa tumor PDX samples collected by the Mayo clinic. Personalized therapy will be designed entailing chemotherapeutic agent(s) and the choice of siRNAs targeting MDR genes. Confirmed platinum-resistant tumor cells will be used to evaluate the anticancer efficacy of NCP/siRNAs in orthotopic PDX OCa mouse models. Tumors from Her-2/neu receptor positive and negative patients will be further evaluated for anticancer efficacy by NCP/siRNAs/H2A. Through these aims we seek to establish a new paradigm for the treatment of resistant OCa. NCP/siRNAs can provide personalized therapy for OCa patients and achieve greatly enhanced anticancer efficacy in resistant OCa. As the standards of care for cervical and other cancers, cisplatin-based NCP/siRNAs can have broad impact on treating other resistant cancers.