Vascular Targeting of Nanocarriers for RNA

用于RNA的脂质纳米颗粒（LNP）的临床批准预示着纳米技术的到来- 基于药物治疗。然而，RNA递送至肝外位点仍然是一个重大的未满足的挑战。 韦斯曼开创了mRNA的修饰，在不同的细胞类型中提供有效的翻译， Muzykantov介绍了“血管靶向”，将药物输送到所需区域的纳米医学策略 在脉管系统中。在这里，我们汇集了这些进展，设计出靶向RNA的纳米载体， 转基因合成和治疗作用的所需位点。我们发现细胞间配体 粘附分子-1（ICAM）与纳米载体结合，指导ICAM-LNP（ILNP）在细胞中积聚， 肺部，尤其是肺部炎症，脑摄取轻微。相反，利比里亚国家警察的目标是 血管细胞粘附分子-1（VCAM）提供VCAM-LNP（VLNP）的轻微肺摄取， 以及炎症大脑的选择性摄取，远远超过了基准。VLNP 载有编码内皮多功能抗血栓和抗炎蛋白的mRNA 血栓调节蛋白（TM）在发炎的CNS中提供转基因合成和保护作用 是其他特工无法比拟的我们将描述并在必要时重新设计目标定位 通过追求以下具体目标，我们将充分利用这一强大的纳米技术平台的特点。目标1： ILNP和VLNP血管靶向的多尺度时空映射。利用同位素示踪 和真实的时间成像，在体内显微镜和流式细胞术，我们将定义PK/BD，LNP的动力学 在期望的治疗作用位点的定位和报告转基因活性。目标二： 将治疗性血栓调节蛋白RNA靶向肺和脑。我们将描述突出参数 TM转基因表达，ILNP和VLNP在幼稚小鼠中的有益和非预期作用， 在肺部和脑部炎症的小鼠模型中。目标3：LNP的翻译发展。 我们将通过位点特异性偶联小的重组蛋白， 配体。这项研究将定义一个新的纳米技术平台的关键规格参数， 选择性地将RNA递送至脑、肺和可能的其他组织中的期望细胞和细胞区室， 机关它将建立一个新的纳米医学方法的有效，具体和 ALI和中风的安全生物药物治疗，未来将扩展到其他疾病。

Clinical approval of Lipid Nano Particles (LNP) for RNA heralded the advent of nanotechnology- based pharmacotherapy. Yet, RNA delivery to extra-hepatic sites remains a major unmet challenge. Weissman pioneered modifications of mRNA providing effective translation in diverse cell types, while Muzykantov introduced “vascular targeting”, nanomedicine strategy for drug delivery to desired areas in the vasculature. Here we converge these advances to devise nanocarriers targeting RNA to desired sites of transgene synthesis and therapeutic action. We found that ligands of Inter-Cellular Adhesion Molecule-1 (ICAM) conjugated to nanocarriers, direct ICAM-LNP (ILNP) to accumulate in lungs, especially, in the inflamed lungs, with trivial cerebral uptake. In opposite, LNP targeting to Vascular Cell Adhesion Molecule-1 (VCAM) provides trivial pulmonary uptake of VCAM-LNP (VLNP), and selective uptake in the inflamed brain, surpassing the benchmarks by orders of magnitude. VLNP loaded with mRNA encoding endothelial multifunctional anti-thrombotic and anti-inflammatory protein thrombomodulin (TM) provides transgene synthesis and protective effect in the inflamed CNS unrivaled by other agents. We will characterize and if needed reiteratively re-engineer targeting features of this powerful nanotechnology platform, by pursuing the following Specific Aims. Aim 1: Multi-scale spatiotemporal mapping of vascular targeting of ILNP and VLNP. Using isotope tracing and real time imaging, in vivo microscopy and flow cytometry, we will define PK/BD, dynamics of LNP localization and reporter transgene activity in the sites of desirable therapeutic action. Aim 2: Targeting therapeutic thrombomodulin RNA to lung and brain. We will characterize salient parameters of TM transgene expression, beneficial and unintended effects of ILNP and VLNP in naive mice and in mouse models of pulmonary and cerebral inflammation. Aim 3: Translational development of LNP. We will upgrade LNP to clinically applicable format via site-specific conjugation of small recombinant ligands. This study will define key specification parameters of a novel nanotechnology platform for selective delivery of RNA to desirable cells and cellular compartments in brain, lungs and likely other organs. It will establish proof of principle for a new nanomedicine approach for effective, specific and safe biological pharmacotherapy of ALI and stroke, with future expansion to other diseases.