Harnessing the power of exosomes for non-coding RNA delivery

项目摘要： 尽管RNA干扰（RNAi）方法有望靶向不可治疗的靶标，但主要挑战是 仍然存在包括siRNA在体内特异性递送到感兴趣的细胞类型中、差的稳定性和脱靶效应。 我们一直处于解决这些问题的最前沿，我的实验室率先进行了许多研究， RNAi方法用于癌症治疗，并取得了与RNAi生物学相关的关键发现。我们 他们是最早证明RNAi加工机制在高比例的卵巢癌中失调的人之一。 和其它癌症（Merritt等人，New Engl J Med 2008）。为了实现RNAi治疗剂的全身递送， 我们系统地鉴定了用于siRNA递送的安全和有效的方法。经过广泛的测试，我们的第一个 成功的平台利用了中性DOPC纳米脂质体递送系统（Landen等人，癌症研究2005; Ahmed等人，Cancer Cell 2010），其随后在多种肿瘤模型系统中进行了测试（Liu et 例如，Nature 2015; Kim等人，Cell 2013）。凭借强大的临床前研究组合和所有必要的 基于FDA指南的安全性研究，EPHARNA（EphA 2靶向）的首次人体I期临床试验 DOPC中的siRNA）用于实体瘤患者的研究已接近完成。我们在应用方面取得了很大的进步 这项技术用于癌症治疗。然而，尽管合成的输送系统，我们和 其他人已经开发了新的和生物相容的递送策略， 血管渗漏是非常需要的。在这个项目中，我们提出了一个仿生外泌体系统， 这将使癌症治疗剂能够主动递送到肿瘤微环境。这些天然存在的 颗粒代表了用于递送RNAi治疗剂的有希望的和安全的替代方法。我们将 将RNAi货物包装到这些颗粒中，并设计它们的表面膜以主动靶向不同的细胞 类型最后，我们将开发这种方法来增强卵巢和其他肿瘤中的抗肿瘤免疫应答。 癌的如果成功，这种仿生外泌体系统可以迅速更广泛地应用于其他生物。 “无法抗拒”的目标。通过R35机制提供的支持将大大促进这项工作， 否则不可能。

Project Abstract: Despite the promise of RNA interference (RNAi) approach for targeting undruggable targets, major challenges remain including specific delivery of siRNA into cell types of interest in vivo, poor stability and off-target effects. We have been at the forefront of addressing these issues and my laboratory has pioneered many studies using RNAi approaches for cancer treatment and has made key discoveries related to RNAi biology. We were among the first to demonstrate that RNAi processing machinery is deregulated in a high proportion of ovarian and other cancers (Merritt et al., New Engl J Med 2008). To achieve systemic delivery of RNAi therapeutics, we systematically identified safe and effective methods for siRNA delivery. After extensive testing, our first successful platform utilized the neutral DOPC nanoliposomal delivery system (Landen et al., Cancer Res 2005; Ahmed et al., Cancer Cell 2010), which has subsequently been tested in multiple tumor model systems (Liu et al., Nature 2015; Kim et al., Cell 2013). With a robust portfolio of preclinical studies and all of the requisite safety studies based on FDA guidance, a first-in-human phase I clinical trial with EPHARNA (EphA2 targeted siRNA in DOPC) is nearing completion for patients with solid tumors. We have made great strides in applying this technology for cancer therapy. However, despite the promise of synthetic delivery systems that we and others have developed, novel and biocompatible delivery strategies that are independent of reliance on vascular leakiness are highly desirable. In this project, we propose to develop a biomimetic exosomal system that will enable active delivery of cancer therapeutics to the tumor microenvironment. These naturally occurring particles represent a promising, and safe alternative approach for delivering RNAi therapeutics. We will package RNAi cargos into these particles and engineer their surface membrane to actively target distinct cell types. Finally, we will develop this approach for enhancing anti-tumor immune response in ovarian and other cancers. If successful, this biomimetic exosomal system can be rapidly applied more broadly to other “undruggable” targets. Support through the R35 mechanism will greatly facilitate this undertaking, which would not otherwise be possible.