In Vivo Pharmacodynamics of RNAi-based Cancer Therapies

由于许多靶点不能用传统的药物化学方法下药，快速开发有效的靶向疗法的困难阻碍了从癌症有效的分子靶标快速转换到使患者受益的治疗。我们在这里提出了一种新的方法，通过使用基于RNA干扰的疗法(这里使用的是基于靶向纳米颗粒的系统)和基于多时间点血液/血清的miRNA图谱，来加速将对癌症细胞生物学中的分子事件的理解转化为新的治疗方法。这一策略将使用以前无法药物治疗的靶点--黑色素瘤中致癌的N-RAS突变进行演示。我们建议评估这样一种假设，即将基于RNA干扰的治疗药物的靶向传递与癌症特异性miRNA生物标记物的多时间点血液/血清监测相结合，可以快速导致通过血液/血清生物标记物进行机械验证的癌症治疗，从而为患者提供微创的功能证据。 具体目标： 1.展示携带siRNA的系统递送纳米粒的肿瘤靶向性、N-RAS基因敲除和抗肿瘤作用。 2.证明了在抑制N-RAS基因产物的RNAi启动后，微创、多时间点血液/血清miRNA测量与肿瘤中多时间点miRNA谱相关。 3.证明系统递送的携带抗N-RAS siRNA的纳米粒可以提供抗肿瘤作用，并且可以通过血液/血清miRNA图谱来监测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