Glycocalyx repair in sepsis using liposomal carriers of preassembled glycocalyx

摘要 富含碳水化合物的最外表层的内皮糖基化蛋白(EG)是一种守护神 血管功能的变化。它是切应力激活的内皮型一氧化氮的传感器 生产和血流诱导的血管扩张，生长因子的先兆，细胞外 超氧化物歧化酶和抗凝血酶III，EG是血管通透性和 白细胞通过血管壁运输，并保护受体免受 通过屏蔽它们来过度刺激。Eg在不同的心血管、代谢中降解 和肾脏疾病，从而导致上述所有功能的损害。这其中的一个 导致EG丢失的条件是败血症，这与高 发病率和死亡率。我们观察到，患有多菌败血症的小鼠表现出 全球EG的数量急剧减少。因此，尽快恢复EG 代表了一种合理的致病疗法。我们最近设计、合成和 在体外、体外和体内测试预组装糖催化剂的脂质体纳米载体。 这种药物干预改善了机械转导和一氧化氮。 内毒素血症小鼠的合成、血流诱导的血管扩张和肾脏微循环。我们 目前已设计并合成了第二代脂质体 以“隐形”脂质体为特征的预组装糖萼纳米载体 半衰期，金标签，以及一系列可能的修改，并演示了 它们显著提高了注射致死剂量的内毒素的小鼠的存活率。这个 本提案旨在探讨该药的有效性和有效性。 脓毒症的手术入路。我们描述了提纯脂质体纳米载体的步骤 预组装糖催化剂，监测这些金标记脂质体的命运-他们的 与质膜融合，细胞内交通，血管系统的半衰期。 之后，我们将确定静脉注射脂质体纳米载体的效果。 脓毒症的病程及与之相关的血流动力学紊乱及其发生率 受影响器官的功能恢复。拟议的研究不仅应该细化这一点 新的治疗工具，但也可能建立一种药理方法来改善 脓毒症引起的多器官衰竭。

ABSTRACT Endothelial glycocalyx (EG), a carbohydrate-rich outermost surface layer, is a guardian of vascular functions. It is a sensor of the shear stress-activated endothelial nitric oxide production and flow-induced vasodilation, harbinger of growth factors, extracellular superoxide dismutase and anti-thrombin III, EG is a regulator of vascular permeability and leukocyte trafficking across the vascular wall, and it protects receptors from hyperstimulation by shielding them. EG is degraded in diverse cardiovascular, metabolic and renal diseases, thus leading to impairment of all the above functions. One of these conditions precipitating the loss of EG is septicemia, which is associated with high morbidity and mortality. We observed that mice with the polymicrobial sepsis exhibit a drastic reduction in the global volume of EG. Therefore, expeditious restoration of EG represents a rational pathogenetic therapy. We have recently designed, synthesized and tested in vitro, ex vivo and in vivo liposomal nanocarriers of preassembled glycocalyx. This pharmacological intervention improved mechanotransduction and nitric oxide synthesis, flow-induced vasodilation and renal microcirculation in endotoxemic mice. We have presently designed and synthesized the second generation of liposomal nanocarriers of preassembled glycocalyx featuring “stealth” liposomes with increased half-life, gold-label, as well as an array of possible modifications and demonstrated that they significantly improved survival of mice injected with the lethal dose of LPS. The present proposal is aimed at exploration of validity and efficacy of this pharmacologic approach in sepsis. We describe the steps to refine liposomal nanocarriers of preassembled glycocalyx, monitoring of the fate of these gold-labeled liposomes – their fusion with the plasma membrane, intracellular traffic, half-life in the vasculature. Thereafter we shall determine the effect of intravenous injection of liposomal nanocarriers on the course of sepsis and associated with it hemodynamic perturbations and the rate of functional restoration of affected organs. Proposed studies should not only refine this novel therapeutic tool but may also establish a pharmacologic approach to ameliorate sepsis-induced multiorgan failure.