Vascular Targeting of Nanocarriers for RNA

RNA 纳米载体的血管靶向

• 批准号: 10093767
• 负责人: Vladimir R Muzykantov
• 金额: $ 72.62万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-05 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10093767
• 关键词: Acute Lung Injury; Affinity; Anti-Inflammatory Agents; Antibodies; Area; Benchmarking; Binding; Biological; Biological Products; Biological Response Modifier Therapy; Blood Vessels; Brain; Cell Adhesion Molecules; Cell membrane; Cells; Cerebrovascular system; Cerebrum; Clinical; Coupled; Development; Disease; Dose; Drug Delivery Systems; Drug Kinetics; Elements; Endocytosis; Endothelial Cells; Endothelium; Engineering; Epitopes; Extrahepatic; Flow Cytometry; Formulation; Functional disorder; Future; Goals; Human; Inflammation; Intercellular adhesion molecule 1; Isotopes; Ligands; Lipids; Liver; Lung; Medical; Messenger RNA; Methods; Microscopy; Modeling; Modification; Mus; Nanotechnology; Organ; Pharmacotherapy; Pilot Projects; Proteins; RNA; RNA delivery; Recombinants; Reporter; Signal Transduction; Site; Spleen; Stroke; Surface; Testing; Therapeutic; Thrombomodulin; Thrombosis; Time; Tissues; Transgenes; Translating; Translations; Vascular Cell Adhesion Molecule-1; base; cell type; clinical application; in vivo imaging; intravenous injection; lead candidate; mouse model; nanobodies; nanocarrier; nanomedicine; nanoparticle; nanotechnology platform; novel; novel strategies; prophylactic; protective effect; real-time images; spatiotemporal; targeted treatment; transgene expression; uptake

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.

用于 RNA 的脂质纳米颗粒 (LNP) 的临床批准预示着纳米技​​术的出现 - 然而，将 RNA 递送至肝外部位仍然是一个尚未解决的重大挑战。 Weissman 率先修饰 mRNA，在不同细胞类型中提供有效的翻译，同时 Muzykantov 推出了“血管靶向”纳米医学策略，将药物输送到所需区域 在这里，我们融合了这些进展来设计靶向 RNA 的纳米载体。 我们发现了细胞间配体的转基因合成和治疗作用的所需位点。 粘附分子 1 (ICAM) 与纳米载体结合，引导 ICAM-LNP (ILNP) 在 肺部，尤其是发炎的肺部，大脑摄取量很少，相反，LNP 的目标是。 血管细胞粘附分子-1 (VCAM) 提供微量肺部摄取 VCAM-LNP (VLNP)， 发炎大脑中的选择性摄取，超过 VLNP 数量级的基准。 装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) 在发炎的中枢神经系统中提供转基因合成和保护作用 我们将描述并在需要时反复重新设计目标。 这个强大的纳米技术平台的特点，通过追求以下具体目标 1： 使用同位素示踪对 ILNP 和 VLNP 的血管靶向进行多尺度时空测绘。 和实时成像、体内显微镜和流式细胞术，我们将定义 PK/BD、LNP 的动态 目标 2：在所需治疗作用位点的定位和报告基因转基因活性。 将治疗性血栓调节蛋白 RNA 靶向肺和脑 我们将表征显着参数。 TM 转基因表达、ILNP 和 VLNP 对幼鼠的有益和意外影响 目标 3：LNP 的转化开发。 我们将通过小重组体的位点特异性缀合将 LNP 升级为临床适用的格式 这项研究将定义新型纳米技术平台的关键规格参数。 将 RNA 选择性递送至脑、肺和其他可能的细胞中所需的细胞和细胞区室 它将为新的纳米医学方法建立有效、具体和有效的原理证明。 ALI 和中风的安全生物药物疗法，未来将扩展到其他疾病。