Systemic RNA Delivery to Tumors

 描述(申请人提供)：鉴于能够调节肿瘤细胞中单个基因的表达，小干扰RNA(SiRNA)和信使RNA(MRNA)等RNA试剂在揭示特定基因改变的功能和使癌症治疗的新型疗法成为可能方面显示出巨大的潜力。然而，对于RNA技术在癌症研究和治疗中的广泛应用来说，有效的系统性RNA传递仍然是一个巨大的挑战。在这个项目中，我们建议(I)开发强大的纳米颗粒(Np)平台，以便在体内有效地将sirna和mrna输送到肿瘤组织中，以及(Ii)将这项技术应用于相关的癌症治疗靶点。 非小细胞肺癌(NSCLC)。在最近的努力中，我们构思和开发了新一代脂质-聚合物杂化纳米粒，具有有望用于全身siRNA传递的特征，包括长血液循环、高肿瘤蓄积和令人印象深刻的基因沉默效果。我们还成功地利用这些杂交纳米粒在非小细胞肺癌中探索了可能的治疗靶点Prohibitin1(PHB1)，并将肿瘤抑制因子编码的mRNA运送到癌细胞，这在体外和体内全身给药后对肿瘤细胞生长的抑制得到了证明。鉴于这些广泛而令人鼓舞的初步结果，我们假设新的脂质-聚合物杂化NP介导的系统RNA递送将成为在体内评估癌症靶点和开发用于癌症治疗的新型RNA疗法的有用平台。在目标1中，我们将系统地优化、了解和评估已被证明控制基因沉默效率和体内特性(例如，药代动力学、生物分布和副作用)的特定参数。在目标2中，我们建议使用来自目标1的优化的siRNA NPs来探索PHB1沉默诱导抗肿瘤作用的机制和途径。用于系统递送抗PHB1 siRNA的杂化NPs将在多个NSCLC异种移植和原位模型中进行广泛的研究。在目标3中，我们将扩展和提炼新的杂交NPs，以提供与NSCLC相关的肿瘤抑制基因的mRNA编码，包括PTEN和LKB1。这种用于癌症治疗的信使核糖核酸传递方法将在体外以及在非小细胞肺癌异种移植和基因工程小鼠模型中进行系统研究。在该项目结束时，我们预计创新的RNA传递策略和肿瘤生物学的重大发现的融合将进一步加强我们对抗非小细胞肺癌和其他侵袭性癌症的治疗手段。

 DESCRIPTION (provided by applicant): Given the capability of modulating individual gene expression in tumor cells, RNA agents such as small interfering RNA (siRNA) and messenger RNA (mRNA) have demonstrated tremendous potential in revealing the functionality of specific gene alterations and enabling novel classes of therapies for cancer treatment. The effective systemic RNA delivery to tumors, however, remains a formidable challenge for the widespread application of RNA technologies in cancer research and therapy. In this project, we propose to (i) develop robust nanoparticle (NP) platforms for effective in vivo delivery of siRNA and mRNA to tumor tissue, and (ii) apply this technology to relevant cancer therapeutic targets for treating non-small cell lung cancer (NSCLC). In recent efforts, we have conceived and developed a new generation of lipid-polymer hybrid NPs with promising features for systemic siRNA delivery, including long blood circulation, high tumor accumulation, and impressive gene silencing efficacy. We have also successfully employed these hybrid NPs to explore a putative therapeutic target, Prohibitin1 (PHB1), in NSCLC and to deliver tumor suppressor-encoded mRNA to cancer cells, as evidenced by inhibition of tumor cell growth in vitro and following systemic delivery in vivo. In light of these extensive and encouraging preliminary results, we hypothesize that the new lipid-polymer hybrid NP-mediated systemic RNA delivery will become a useful platform for in vivo evaluation of cancer targets and for the development of novel RNA therapies for cancer treatment. In Aim 1, we will systematically optimize, understand and evaluate specific parameters that have been shown to control gene silencing efficiency and in vivo properties (e.g., pharmacokinetics, biodistribution, and side effects) of the hybrid NPs. In Aim 2, we propose to explore the mechanisms and pathways underlying the PHB1 silencing-induced anti-tumor effect, using the optimized siRNA NPs from Aim 1. The hybrid NPs for systemic delivery of anti-PHB1 siRNA will be extensively examined in multiple NSCLC xenograft and orthotopic models. In Aim 3, we will expand and refine the new hybrid NPs to deliver mRNA coding for tumor suppressors relevant to NSCLC, including PTEN and LKB1. This mRNA delivery approach for cancer therapy will be systematically investigated in vitro and in NSCLC xenograft and genetically engineered mouse models. At the conclusion of this project, we expect that the convergence of innovative RNA delivery strategy and significant discoveries in tumor biology will further strengthen our arsenal of therapies against NSCLC and other aggressive cancers.