RNA Nanoparticles-Displaying Exosomes to enhance targeted siRNA delivery for Prostate Cancer treatment.

项目概要 前列腺癌是男性中最常见的癌症类型，也是第六大癌症原因 每年都有死亡。虽然临床管理可采用不同的治疗策略 疾病（手术、放射治疗、化疗、激素治疗）研究正在转向创新 基于 RNA 干扰剂递送的方法，包括小干扰 RNA (siRNA)。 尽管基于 siRNA 的策略能够有效促进肿瘤抑制，但它们仍受到以下因素的负面影响： siRNA 的递送不良、体内半衰期短以及免疫反应的改变，这些都阻碍了它们的应用 转化为临床实践。外泌体已被提议作为递送纳米粒子，这要归功于其 自然参与蛋白质和核酸的细胞间运输。然而，外泌体遭受 技术限制（靶向效率低、生物分布差、内体捕获）。外显子RNA 提出了一种基于 RNA 纳米技术来生成 RNA 纳米颗粒的新方法-显示 用于在前列腺癌细胞中靶向递送 siRNA 的外泌体。 RNA纳米粒子的用途 与外泌体相关可以实现卓越的细胞靶向性和 siRNA 的有效递送。外泌体是 经改造，显示出良好的药代动力学特征和低免疫原性，并避免内体 诱捕。该公司已初步验证使用纳米颗粒定向来控制RNA加载 在一项支持 RNA 纳米颗粒使用的概念验证研究中，外泌体上的配体展示 加载外泌体来靶向前列腺癌。在这个 SBIR 一期项目中，ExonanoRNA 将产生先导物 具有高水平的稳定性、生产效率、治疗效果和安全性的候选者，适合 临床测试。这一目标将通过两个目标来实现。目标1）选择最合适的 靶向配体以确保对前列腺细胞的最大特异性并确定最佳数量 与 EV 相关的 RNA NP 以实现最有效的靶向效率。目标2）最好的 候选者将在人前列腺腺癌细胞（LNCaP）上进行体外测试，并在体内进行体内测试 异种移植小鼠以研究功效和安全性。拟议的活动将导致第二阶段项目 其中 ExonanoRNA 预计扩大已验证候选药物的生产规模并执行 IND- 能够对小型和大型动物（大鼠、比格犬）进行研究。

PROJECT SUMMARY Prostate Cancer is the most common type of cancer diagnosed in the male population and the sixth cause of death every year. While different therapeutic strategies are available for the clinical management of the disease (surgery, radiation therapy, chemotherapy, hormonal therapy) research is shifting to innovative approaches based on the delivery of RNA interfering agents, including small interfering RNA (siRNA). Despite their efficacy in promoting tumor suppression, siRNA-based strategies are negatively affected by poor delivery of siRNA, short half-life in vivo, and alteration of the immune response, which hinder their translation into clinical practice. Exosomes have been proposed as delivery nanoparticles thanks to their natural involvement in intercellular transport of protein and nucleic acids. However, exosomes suffer from technical limitations (low targeting efficiency, poor biodistribution, endosome trapping). ExonanoRNA proposes a novel approach based on RNA Nanotechnology to generate RNA Nanoparticles-Displaying Exosomes for the targeted delivery of siRNA in prostate cancer cells. The use of RNA nanoparticles associated with exosomes allows superior cell targeting and efficient delivery of siRNA. Exosomes are engineered to display a favorable pharmacokinetic profile and low immunogenicity and to avoid endosome trapping. The company has preliminary validated the use of nanoparticle orientation to control RNA loading and ligand display on exosomes in a proof-of-concept study that supports the use of RNA nanoparticles- loaded Exosomes to target prostate cancer. In this SBIR Phase I project, ExonanoRNA will generate lead candidates with high level of stability, production efficiency, therapeutic efficacy, and safety suitable for clinical testing. This goal will be achieved through two aims. Aim 1) selection of the most appropriate targeting ligand to ensure the maximum specificity to prostate cells and identification of the optimal number of RNA NPs associated to the EVs to achieve the most efficient targeting efficiency. Aim 2) The best candidate will be tested in vitro on human prostate adenocarcinoma (LNCaP+) cells, and in vivo on xenografts mice to investigate efficacy, and safety. The proposed activities will lead to a Phase II project where ExonanoRNA foresees the scale up of production of the validated candidate and perform IND- enabling studies in small and large animals (rat, beagle dog).