In Vivo Pharmacodynamics of RNAi-based Cancer Therapies

从癌症中经过验证的分子靶标到使患者受益的治疗的快速转化受到快速开发有效的靶向治疗方法的困难的阻碍，因为许多靶标不能通过传统的药物化学方法进行药物化。我们在这里提出了一种新的方法，通过使用RNA干扰为基础的治疗（靶向纳米粒子为基础的系统在这里使用）结合多时间点血液/血清为基础的miRNA分析，加快翻译的理解在癌细胞生物学的分子事件到新的疗法。这一策略将证明使用以前undruggable目标，致癌N-Ras突变在黑色素瘤。我们建议评估这样的假设，即基于RNA干扰的治疗的靶向递送的整合，允许从概念到临床的快速转化与癌症特异性miRNA生物标志物的多时间点血液/血清监测，可以快速导致使用血液/血清生物标志物的机械验证的癌症治疗，可以提供患者的微创功能证明。 具体目标： 1.展示携带siRNA的全身递送纳米颗粒的肿瘤靶向、N-Ras敲低和抗肿瘤作用。 2.证明在启动抑制N-Ras基因产物的RNAi后，微创、多时间点血液/血清miRNA测量与肿瘤中的多时间点miRNA谱相关。 3.证明携带抗N-Ras siRNA的全身递送的纳米颗粒可以提供抗肿瘤作用，并且可以通过血液/血清miRNA谱监测N-Ras基因抑制的时间过程。

Rapid translation from validated molecular targets in cancer to treatments that benefit patients has been hampered by difficulties in rapidly developing effective, targeted therapeutics, as many targets are not druggable by traditional medicinal chemistry approaches. We propose here to invesfigate a new methodology to accelerate the translation of understandings of molecular events in cancer cell biology into new therapies by using RNA interference-based therapeutics (targeted nanoparticle based systems are used here) combined with multi-time point blood/serum-based miRNA profiling. This strategy will be demonstrated using a previously undruggable target, the oncogenic N-Ras mutation in melanoma. We propose to evaluate the hypothesis that the integration of targeted delivery of RNA interference-based therapeutics that allow quick translation from concept to clinic with multi-time point blood/serum monitoring of cancer-specific miRNA biomarkers can rapidly lead to mechanistically verified cancer treatments with blood/serum biomarkers that can provide minimally invasive proof of function in patients. Specific Aims: 1. Demonstrate tumor targeting, N-Ras knock-down, and anti-tumor effects with systemically delivered nanoparticles carrying siRNA. 2. Demonstrate that minimally invasive, multi-time point blood/serum miRNA measurements correlate with multi-time point miRNA profiles in tumors after initiation of RNAi that inhibits the N-Ras gene product. 3. Demonstrate that systemically delivered nanoparticles carrying anti-N-Ras siRNA can provide antitumor effects and that the time course of the N-Ras gene inhibition can be monitored via blood/serum miRNA profiles