Augmentation of Antioxidant Defenses by Immunotargeting

通过免疫靶向增强抗氧化防御

• 批准号: 6780632
• 负责人: Vladimir R Muzykantov
• 金额: $ 39.63万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6780632
• 关键词: antioxidants; bioengineering /biomedical engineering; catalase; cell adhesion molecules; cytoprotection; disease /disorder model; drug delivery systems; drug design /synthesis /production; drug screening /evaluation; free radical oxygen; gene deletion mutation; hydrogen peroxide; hyperoxia; immunoconjugates; intracellular transport; laboratory mouse; laboratory rat; lung injury; lysosomes; oxidative stress; pharmacokinetics; protein transport; sodium hydrogen exchanger; superoxide dismutase; tissue /cell culture; transfection; vascular endothelium

DESCRIPTION (provided by applicant): Vascular oxidative stress is a key component of many lung diseases, but its treatment is inadequate, in part due to sub-optimal delivery of antioxidants to endothelial cells (EC). Previous studies have shown that: 1) antibodies to Platelet Endothelial Adhesion Molecule (PECAM) permit intracellular delivery of drugs to EC via a novel endocytotic pathway distinct from clathrin- and caveoli-mediated endocytosis; and 2) the antioxidant enzyme (AOE) catalase conjugated with anti-PECAM accumulates in the lungs after IV injection and protects against oxidative lung injury in some animal models. The aim of this translational grant is to enhance the duration and effectiveness of protective effects of this promising strategy to clinically significant levels. We hypothesize that: i) the activity of the conjugates can be prolonged by optimization of their design and by manipulating intracellular trafficking and lysosomal degradation; ii) targeted delivery of additional AOEs to EC will permit enhanced protection from oxidative stress; and iii) pulmonary targeting of optimized AOE conjugates will be protective against hyperoxia. We will test this in the following Specific Aims: 1) define the mechanisms of metabolism of anti-PECAM conjugates in EC. We will test the hypothesis that endocytosis and trafficking involve the cytosolic domain of PECAM, Na+-H+ exchangers (NHE) and rearrangements of cytoskeleton, which can be affected by auxiliary agents to prolong duration of conjugates; 2) design targeting of additional AOE. To enhance protection, tandem SOD/catalase (to detoxify 02- and H202) and 1-CysPrx peroxiredoxin (to detoxify H202 and lipid peroxides) conjugates will be produced. Their composition, activities, EC uptake and protection will be studied in cell culture, while their pharmacokinetics and pulmonary targeting will be determined in naive animals and animals with oxidative lung injury; and 3) evaluate protective effects of the conjugates in animal models. The effectiveness and duration of the effects of new conjugates will be compared and regimens of AOE targeting and mechanisms of protection will be studied in a model of acute EC injury induced by H202 in the pulmonary vasculature in mice. Finally, protection against hyperoxic lung injury by best conjugates will be tested. The overall goal of this proposal is to optimize strategies for vascular AOE targeting, to ultimately initiate translation of this strategy into the clinical domain.

描述（由申请人提供）：血管氧化应激是许多肺部疾病的关键组成部分，但其治疗是不够的，部分原因是抗氧化剂向内皮细胞（EC）的次优递送。先前的研究已经表明：1）血小板内皮粘附分子（PECAM）的抗体允许药物通过不同于网格蛋白和小窝介导的内吞作用的新型内吞途径细胞内递送至EC;和2）与抗PECAM缀合的抗氧化酶（AOE）过氧化氢酶在IV注射后在肺中积累，并在一些动物模型中保护免受氧化性肺损伤。该翻译补助金的目的是将这种有前途的策略的保护作用的持续时间和有效性提高到临床显著水平。我们假设：i）缀合物的活性可以通过优化其设计和通过操纵细胞内运输和溶酶体降解来延长; ii）将另外的AOE靶向递送至EC将允许增强对氧化应激的保护;和iii）优化的AOE缀合物的肺靶向将保护免于高氧。 我们将在以下具体目的中对此进行测试：1）定义抗PECAM缀合物在EC中的代谢机制。 我们将检验以下假设：内吞作用和运输涉及PECAM的胞质结构域、Na+-H+交换剂（NHE）和细胞骨架的重排，这可以受到辅助剂的影响以延长缀合物的持续时间; 2）设计额外AOE的靶向。为了增强保护，将产生串联SOD/过氧化氢酶（使O2-和H2 O2解毒）和1-CysPrx过氧化物氧还蛋白（使H2 O2和脂质过氧化物解毒）缀合物。将在细胞培养中研究它们的组成、活性、EC摄取和保护，同时将在未处理动物和具有氧化性肺损伤的动物中确定它们的药代动力学和肺靶向;和3）评估缀合物在动物模型中的保护作用。将比较新缀合物的效果的有效性和持续时间，并将在小鼠肺血管中由H2 O2诱导的急性EC损伤模型中研究AOE靶向方案和保护机制。 最后，将测试最佳缀合物对高氧肺损伤的保护作用。该提案的总体目标是优化血管AOE靶向策略，最终将该策略转化为临床领域。