Greineder Colin F, Brenza Jacob B, Carnemolla Ronald, Zaitsev Sergei, Hood Elizabeth D, Pan Daniel C, Ding Bi-Sen, Esmon Charles T, Chacko Ann Marie, Muzykantov Vladimir R
*Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA.
*Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA
FASEB J. 2015 Aug;29(8):3483-92. doi: 10.1096/fj.15-271213. Epub 2015 May 7.
Anchoring pharmacologic agents to the vascular lumen has the potential to modulate critical processes at the blood-tissue interface, avoiding many of the off-target effects of systemically circulating agents. We report a novel strategy for endothelial dual targeting of therapeutics, which both enhances drug delivery and enables targeted agents to partner enzymatically to generate enhanced biologic effect. Based on the recent discovery that paired antibodies directed to adjacent epitopes of platelet endothelial cell adhesion molecule (PECAM)-1 stimulate each other's binding, we fused single-chain fragments (scFv) of paired anti-mouse PECAM-1 antibodies to recombinant murine thrombomodulin (TM) and endothelial protein C receptor (EPCR), endothelial membrane proteins that partner in activation of protein C (PC). scFv/TM and scFv/EPCR bound to mouse endothelial PECAM-1 with high affinity (EC50 1.5 and 3.8 nM, respectively), and codelivery induced a 5-fold increase in PC activation not seen when TM and EPCR are anchored to distinct cell adhesion molecules. In a mouse model of acute lung injury, dual targeting reduces both the expression of lung inflammatory markers and trans-endothelial protein leak by as much as 40%, as compared to either agent alone. These findings provide proof of principle for endothelial dual targeting, an approach with numerous potential biomedical applications.
将药理剂锚定到血管腔有潜力调节血液-组织界面的关键过程，避免全身循环药剂的许多脱靶效应。我们报告了一种用于治疗剂内皮双靶向的新策略，该策略既能增强药物递送，又能使靶向药剂通过酶促作用协同产生增强的生物学效应。基于最近的发现，即针对血小板内皮细胞粘附分子（PECAM）-1相邻表位的配对抗体相互刺激彼此的结合，我们将配对抗小鼠PECAM-1抗体的单链片段（scFv）与重组小鼠血栓调节蛋白（TM）和内皮蛋白C受体（EPCR）融合，这两种内皮膜蛋白在蛋白C（PC）激活中协同作用。scFv/TM和scFv/EPCR以高亲和力（分别为EC50 1.5和3.8 nM）与小鼠内皮PECAM-1结合，与将TM和EPCR锚定到不同细胞粘附分子时相比，共递送诱导PC激活增加了5倍。在急性肺损伤小鼠模型中，与单独使用任何一种药剂相比，双靶向可使肺部炎症标志物的表达和跨内皮蛋白渗漏减少多达40%。这些发现为内皮双靶向提供了原理证明，这是一种具有众多潜在生物医学应用的方法。
