Vascular Targeting of Nanocarriers for RNA

脂质纳米颗粒(LNP)用于RNA的临床批准预示着纳米技术的到来-- 以药物治疗为基础。然而，将RNA运送到肝外部位仍然是一个尚未满足的主要挑战。 魏斯曼率先对信使核糖核酸进行修饰，在不同的细胞类型中提供了有效的翻译，而 Muzykantov介绍了“血管靶向”，这是一种将药物输送到目标区域的纳米药物战略 在血管系统中。在这里，我们汇集了这些进展，以设计针对RNA的纳米载体来 转基因合成和治疗作用的理想部位。我们发现细胞间的配体 黏附分子-1(ICAM)偶联到纳米载体上，引导ICAM-LNP(ILNP)在 肺部，特别是发炎的肺部，有少量的大脑摄取。相反，LNP的目标是 血管细胞黏附分子-1(VCAM)提供对VCAM-LNP(VLNP)的少量肺摄取， 在发炎的大脑中选择性摄取，超过基准数量级。VLNP 携带编码血管内皮细胞多功能抗血栓抗炎蛋白的mRNA 血栓调节蛋白(TM)在炎症的中枢神经系统中提供转基因合成和保护作用 是其他代理商所无法比拟的。我们将确定目标的特征，并在需要时反复重新设计目标 这一强大的纳米技术平台的特点，通过追求以下具体目标。目标1： ILNP和VLNP血管靶向的多尺度时空标测。使用同位素示踪 实时成像，活体显微镜和流式细胞术，我们将定义PK/BD，LNP的动力学 在理想的治疗作用部位的定位和报告转基因活性。目标2： 靶向肺和脑的治疗性血栓调节蛋白RNA。我们将描述显著参数 对转基因表达的影响，ILNP和VLNP对幼鼠和 在小鼠的肺部和脑部炎症模型中。目标3：LNP的翻译发展。 我们将通过小重组蛋白的定点结合将LNP升级为临床适用的形式 配基。这项研究将确定一种新型纳米技术平台的关键规格参数 选择性地将RNA递送到所需的细胞和脑、肺以及可能的其他组织中的细胞室 器官。它将为一种新的纳米医学方法建立原则证明，以实现有效、特异和 ALI和卒中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.