Vascular delivery of nanocarriers by erythrocyres

项目摘要 血液和组织之间的多功能内皮界面是免疫调节的重要靶点。 对许多人类疾病的治疗干预。为了实现精确的干预，许多实验室 包括US缀合药物和具有将货物靶向至 内皮细胞另一方面，通过非亲和性机制在组织中积累的载体可能 提供药物递送能力的额外提升。我们已经发现， 纳米载体（NC）与红细胞（RBC）表面的结合提供了一种新的策略， 和非针对性的方法。吸附在分离的RBC上的NC（RBC/NC）迅速转移到 注射部位下游的血管系统并避免肝脏吸收。试点数据显示，我们可以 协同RBC搭便车和亲和力靶向的力量。在RBC上加载几乎提供了 肺中EC靶向NC的摄取增加三个数量级。此外，RBC靶向 在体内，NCs安全地加载到RBC上，这使我们能够避免输血。将这些联合收割机 优势，并使从红细胞转移到EC，我们设计了双靶向NC（DTNC） 通过与结合RBC和EC的各向异性“Janus”颗粒配体的相对面缀合。 每个方面的亲合力的微调最大化靶向RBC和转移的时空控制 到EC。我们鉴定了选择性地将NC靶向脑与肺的配体。这项提案的目的是 是定义机制，并使这种新颖的，范式转换策略的翻译成为可能。我们将 采用相互增强的模型：体外（微流体）、离体（灌注人肺）和体内 (na病理动物）。我们将研究NC加载到RBC上，并转移到 在受体细胞中的定位，以及在三个独立的细胞中通过RBC搭便车的药物递送的作用。 目标。目的1：将NC加载到RBC上。我们将：A）定义RBC加载的最佳NC设计; B） 在体内工程化RBC靶向NC负载;以及，C）确定NC负载的生物相容性。 红细胞。目标2：NC卸载和转移。我们将描述和优化血管转移， 非靶向NC与EC靶向和双重靶向NC：A）转移的动力学和幅度; B） NC的细胞寻址和运输;和，C）调节转移的病理生理因素。 目标3：翻译RBC搭便车。我们将：A）评估有益与非预期的影响 通过RBC/NC递送抗炎剂; B）使NC靶向人RBC;以及C） 概述在灌注的人肺中进行的动物研究的主要发现。本研究将推进：A） 结合靶向纳米载体与“超载体”RBC的药物递送系统的设计; B） 了解重要的血管界面; C）开发精确靶向的 用于治疗ALI/ARDS和可能的中风和其他常见急性危象的药物疗法。

Project Summary The multifunctional endothelial interface between blood and tissues is an important target for therapeutic interventions in many human maladies. To achieve precise interventions, many labs including us conjugate drugs and drug carriers with affinity ligands that target cargoes to the endothelium. On the other hand, carriers that accumulate in tissues via non-affinity mechanisms may provide an additional boost in drug delivery capacity. We have found that reversible association of nanocarriers (NCs) with the red blood cell (RBC) surface provides a new strategy combining targeted and non-targeted approaches. NCs adsorbed onto isolated RBCs (RBC/NC) rapidly transfer to the vasculature downstream of the injection site and avoid hepatic uptake. Pilot data show that we can synergize the power of RBC-hitchhiking and affinity targeting. Loading on RBCs provides almost three orders of magnitude boost of uptake of EC-targeted NCs in the lungs. Further, RBC-targeted NCs safely load onto RBCs in vivo, which allows us to avoid transfusion. To combine these advantages and enable transfer from RBCs to ECs, we have designed dual-targeted NCs (DTNCs) by conjugating to opposite facets of anisotropic “Janus” particles ligands that bind to RBCs and EC. Fine-tuning of each facet's avidity maximizes spatiotemporal control of targeting to RBCs and transfer to ECs. We identified ligands selectively targeting NCs to the brain vs lungs. The goal of this proposal is to define the mechanism and enable translation of this novel, paradigm-shifting strategy. We will employ mutually reinforcing models: in vitro (microfluidic), ex vivo (perfused human lungs) and in vivo (naïve vs pathological animals). We will study NC loading onto RBC and the transfer to and localization in recipient cells, and the effect of drug delivery by RBC-hitchhiking in three independent Aims. Aim 1: Loading NC onto RBCs. We will: A) Define optimal NC design for RBC loading; B) Engineer RBC-targeted NC loading in vivo; and, C) Determine the biocompatibility of NC-loaded RBCs. Aim 2: NC unloading and transfer. We will characterize and optimize vascular transfer of untargeted NCs vs EC-targeted and dual-targeted NCs: A) Kinetics and amplitude of transfer; B) Cellular addressing and trafficking of NCs; and, C) Pathophysiological factors modulating transfer. Aim 3: Translational RBC hitchhiking. We will: A) Appraise beneficial vs unintended effects of delivery of anti-inflammatory agents by RBC/NC; B) Refine NC targeting to human RBC; and, C) Recapitulate key findings of animal studies in perfused human lungs. This study will advance: A) Design of drug delivery systems combining targeted nanocarriers with “supercarrier” RBCs; B) Understanding of important vascular interfaces; C) Development of precisely targeted pharmacotherapy for treatment of ALI/ARDS and likely stroke and other common acute crises.