Development of PolyAspirin Particles for Therapeutic Intervention in ALI/ARDS via the Passive Restraint of Neutrophil Function

摘要 这项工作的中心目标是开发静脉注射(IV)的聚阿司匹林(Poly-A)颗粒作为 急性肺损伤(ALI)治疗干预中对中性粒细胞功能的被动抑制阿里是一个迅速发展的 以肺内皮细胞和上皮屏障破坏为特征的进行性炎症性疾病， 导致液体在肺部呼吸道积聚，从而损害肺功能。阿里与急性 呼吸窘迫综合征(ARDS)是一种更严重的ALI，在美国每年影响约20万名患者 目前，死亡率约为40%-60%。到目前为止，还没有一种有效的药物策略来治疗 降低ALI/ARDS的死亡率，可能是由于大量和复杂的病理事件集 导致了这种疾病。因此，这种疾病的主要治疗方法是使用机械换气机来换血。 氧合和二氧化碳清除使受损的肺得以愈合，但这可能会导致对肺的进一步损害 如果不小心使用的话。中性粒细胞已被确定为炎症的主要肇事者 ALI/ARDS，它们过度向肺内迁移导致肺泡毛细血管的破坏 导致肺部浮肿的屏障。事实上，疾病的严重程度与中性粒细胞的浓度有关。 在肺部的呼吸道里。因此，阻止有害的中性粒细胞聚集的破坏性潜力一直是一个 ALI/ARDS治疗的发展重点。然而，之前开发药物的尝试 由于大量的冗余，阻断中性粒细胞信号/黏附分子的成功有限。 在炎症反应级联反应中。在这里，我们建议合理地设计血管靶向颗粒(VTP) 与中性粒细胞发生物理作用，被动而快速地阻止中性粒细胞聚集到炎症组织 在ALI/ARDS。我们的主要假设是VTP与血液中的中性粒细胞相互作用，通过物理 相互作用和竞争血管结合空间，改变中性粒细胞与血管壁的黏附，这是 严重影响它们向病变组织的迁移。在本提案中，我们利用这些阻塞交互 目的：通过三个靶点开发一种可生物降解的、生物相容的VTPS，作为治疗ALI/ARDS的有效药物。 首先，我们将构建一种基于聚阿司匹林的VTP系统，并评估其颗粒大小和表面的影响 它们在体外特异性阻断中性粒细胞与血管壁黏附的能力。第二，我们将 通过活体显微镜成像，可视化基于聚阿司匹林的VTP与血管壁的粘附性 以及它们在体内阻断炎症小鼠肠系膜组织中的中性粒细胞黏附。第三，我们将评估 聚阿司匹林颗粒对细菌诱导的ALI/ARDS小鼠的治疗作用 人类疾病的真实模型。总体而言，从这些目标获得的知识预计将推动 新型颗粒型抗炎药物在ALI/ARDS治疗中的应用前景这个 所提出的VTP对中性粒细胞的直接作用，而不是阻断黏附或信号分子，将 确保无论ALI/ARDS的主要原因是什么，建议的系统都能正常运行。

ABSTRACT The central goal of this work is to develop intravenously (IV)-injected Poly-Aspirin (Poly-A) particles as passive restraints of neutrophil function for therapeutic intervention in Acute Lung Injury (ALI). ALI is a rapidly progressing inflammatory disease characterized by the disruption of the lung endothelial and epithelial barriers, leading to accumulation of fluids in the lung airway and hence impaired lung function. ALI together with acute respiratory distress syndrome (ARDS), a more severe form of ALI, affects ~200,000 patients per year in the US currently, with a mortality rate of ~40-60%. To date, there is no one pharmacological strategy effective towards reducing the mortality in ALI/ARDS, likely due to the numerous and complex set of pathological events that can lead to this disease. Thus, the primary treatment for this disease is the use of a mechanical ventilator for blood oxygenation and CO2 removal to allow the damaged lung to heal, but this can lead to further damage to the lung if not employed with care. Neutrophils have been identified as the primary perpetrator of inflammation in ALI/ARDS, where their excessive migration into the lungs contributes to the destruction of the alveolar-capillary barrier that leads to edema in the lungs. Indeed, disease severity correlates with the concentration of neutrophils in the lung airways. Thus, halting the destructive potential of unwanted neutrophil accumulation has been a principal focus for the development of ALI/ARDS treatment. However, prior attempts at developing drugs that block neutrophil signaling/adhesion molecules have met with limited success due to the numerous redundancies in the inflammatory response cascade. Here, we propose to rationally design vascular-targeted particles (VTPs) that physically interact with neutrophils to passively and rapidly block neutrophil accumulation into inflamed tissue in ALI/ARDS. Our main hypothesis is that VTPs interact with neutrophils in the bloodstream, via physical interaction and competition for vascular binding space, to alter neutrophil adhesion to the vessel wall, which critically impacts their migration into the diseased tissue. In this proposal, we harness these blocking interactions to develop a biodegradable, biocompatible VTPs as an effective treatment for ALI/ARDS through three Aims. First, we will fabricate a PolyAspirin-based VTP system and evaluate the impact of their particle size and surface characteristics on their ability to specifically block neutrophil adhesion to the vessel wall in vitro. Second, we will visualize, via intravital microscopy imaging, the adhesion of the PolyAspirin-based VTPs to the blood vessel wall and their blocking of neutrophil adhesion in vivo in inflamed mesentery tissue in mice. Thirdly, we will evaluate the therapeutic functionality of PolyAspirin particles in mice with bacteria-induced ALI/ARDS, representing a realistic model of the human disease. Overall, the knowledge gained from these Aims is expected to drive the future development of novel particle-based anti-inflammatory therapeutics in the treatment of ALI/ARDS. The proposed direct action of VTPs on neutrophils, rather than blocking of adhesion or signaling molecules, will ensure that the proposed system can function irrespective of the primary cause of ALI/ARDS.