Use of Electron Paramagnetic Resonance (EPR) to Evaluate Redox Status in a Preclinical Model of Acute Lung Injury.

IF 3 4区医学 Q2 RADIOLOGY, NUCLEAR MEDICINE & MEDICAL IMAGING Molecular Imaging and Biology Pub Date : 2024-06-01 Epub Date: 2023-05-16 DOI:10.1007/s11307-023-01826-5

Hanan B Elajaili, Nathan M Dee, Sergey I Dikalov, Joseph P Y Kao, Eva S Nozik

{"title":"Use of Electron Paramagnetic Resonance (EPR) to Evaluate Redox Status in a Preclinical Model of Acute Lung Injury.","authors":"Hanan B Elajaili, Nathan M Dee, Sergey I Dikalov, Joseph P Y Kao, Eva S Nozik","doi":"10.1007/s11307-023-01826-5","DOIUrl":null,"url":null,"abstract":"Purpose: Patients with hyper- vs. hypo-inflammatory subphenotypes of acute respiratory distress syndrome (ARDS) exhibit different clinical outcomes. Inflammation increases the production of reactive oxygen species (ROS) and increased ROS contributes to the severity of illness. Our long-term goal is to develop electron paramagnetic resonance (EPR) imaging of lungs in vivo to precisely measure superoxide production in ARDS in real time. As a first step, this requires the development of in vivo EPR methods for quantifying superoxide generation in the lung during injury, and testing if such superoxide measurements can differentiate between susceptible and protected mouse strains.Procedures: In WT mice, mice lacking total body extracellular superoxide dismutase (EC-SOD) (KO), or mice overexpressing lung EC-SOD (Tg), lung injury was induced with intraperitoneal (IP) lipopolysaccharide (LPS) (10 mg/kg). At 24 h after LPS treatment, mice were injected with the cyclic hydroxylamines 1-hydroxy-3-carboxy-2,2,5,5-tetramethylpyrrolidine hydrochloride (CPH) or 4-acetoxymethoxycarbonyl-1-hydroxy-2,2,5,5-tetramethylpyrrolidine-3-carboxylic acid (DCP-AM-H) probes to detect, respectively, cellular and mitochondrial ROS - specifically superoxide. Several probe delivery strategies were tested. Lung tissue was collected up to one hour after probe administration and assayed by EPR.Results: As measured by X-band EPR, cellular and mitochondrial superoxide increased in the lungs of LPS-treated mice compared to control. Lung cellular superoxide was increased in EC-SOD KO mice and decreased in EC-SOD Tg mice compared to WT. We also validated an intratracheal (IT) delivery method, which enhanced the lung signal for both spin probes compared to IP administration.Conclusions: We have developed protocols for delivering EPR spin probes in vivo, allowing detection of cellular and mitochondrial superoxide in lung injury by EPR. Superoxide measurements by EPR could differentiate mice with and without lung injury, as well as mouse strains with different disease susceptibilities. We expect these protocols to capture real-time superoxide production and enable evaluation of lung EPR imaging as a potential clinical tool for subphenotyping ARDS patients based on redox status.","PeriodicalId":18760,"journal":{"name":"Molecular Imaging and Biology","volume":" ","pages":"495-502"},"PeriodicalIF":3.0000,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10188229/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular Imaging and Biology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1007/s11307-023-01826-5","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/5/16 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"RADIOLOGY, NUCLEAR MEDICINE & MEDICAL IMAGING","Score":null,"Total":0}

引用次数: 0

Abstract

Purpose: Patients with hyper- vs. hypo-inflammatory subphenotypes of acute respiratory distress syndrome (ARDS) exhibit different clinical outcomes. Inflammation increases the production of reactive oxygen species (ROS) and increased ROS contributes to the severity of illness. Our long-term goal is to develop electron paramagnetic resonance (EPR) imaging of lungs in vivo to precisely measure superoxide production in ARDS in real time. As a first step, this requires the development of in vivo EPR methods for quantifying superoxide generation in the lung during injury, and testing if such superoxide measurements can differentiate between susceptible and protected mouse strains.

Procedures: In WT mice, mice lacking total body extracellular superoxide dismutase (EC-SOD) (KO), or mice overexpressing lung EC-SOD (Tg), lung injury was induced with intraperitoneal (IP) lipopolysaccharide (LPS) (10 mg/kg). At 24 h after LPS treatment, mice were injected with the cyclic hydroxylamines 1-hydroxy-3-carboxy-2,2,5,5-tetramethylpyrrolidine hydrochloride (CPH) or 4-acetoxymethoxycarbonyl-1-hydroxy-2,2,5,5-tetramethylpyrrolidine-3-carboxylic acid (DCP-AM-H) probes to detect, respectively, cellular and mitochondrial ROS - specifically superoxide. Several probe delivery strategies were tested. Lung tissue was collected up to one hour after probe administration and assayed by EPR.

Results: As measured by X-band EPR, cellular and mitochondrial superoxide increased in the lungs of LPS-treated mice compared to control. Lung cellular superoxide was increased in EC-SOD KO mice and decreased in EC-SOD Tg mice compared to WT. We also validated an intratracheal (IT) delivery method, which enhanced the lung signal for both spin probes compared to IP administration.

Conclusions: We have developed protocols for delivering EPR spin probes in vivo, allowing detection of cellular and mitochondrial superoxide in lung injury by EPR. Superoxide measurements by EPR could differentiate mice with and without lung injury, as well as mouse strains with different disease susceptibilities. We expect these protocols to capture real-time superoxide production and enable evaluation of lung EPR imaging as a potential clinical tool for subphenotyping ARDS patients based on redox status.

Abstract Image

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

利用电子顺磁共振 (EPR) 评估急性肺损伤临床前模型的氧化还原状态

目的：急性呼吸窘迫综合征（ARDS）的高炎症亚型和低炎症亚型患者表现出不同的临床结局。炎症会增加活性氧（ROS）的产生，而 ROS 的增加会加重病情。我们的长期目标是开发体内肺部电子顺磁共振（EPR）成像技术，以实时精确测量 ARDS 中超氧化物的产生。作为第一步，我们需要开发体内电子顺磁共振方法来量化损伤过程中肺部产生的超氧化物，并测试这种超氧化物测量是否能区分易感和受保护的小鼠品系：在 WT 小鼠、缺乏全身细胞外超氧化物歧化酶（EC-SOD）（KO）或过表达肺 EC-SOD （Tg）的小鼠中，用腹腔注射（IP）脂多糖（LPS）（10 毫克/千克）诱导肺损伤。LPS 处理 24 小时后，给小鼠注射环羟胺 1-羟基-3-羧基-2,2,5,5-四甲基吡咯烷盐酸盐（CPH）或 4-乙酰氧基甲氧基羰基-1-羟基-2,2,5,5-四甲基吡咯烷-3-羧酸（DCP-AM-H）探针，分别检测细胞和线粒体 ROS（特别是超氧化物）。对几种探针递送策略进行了测试。在使用探针一小时后收集肺组织，并通过 EPR 进行检测：通过 X 波段 EPR 测定，与对照组相比，经 LPS 处理的小鼠肺部细胞和线粒体超氧化物增加。与 WT 小鼠相比，EC-SOD KO 小鼠肺细胞超氧化物增加，而 EC-SOD Tg 小鼠肺细胞超氧化物减少。我们还验证了气管内给药方法，与 IP 给药相比，该方法增强了两种自旋探针的肺部信号：结论：我们制定了在体内递送 EPR 自旋探针的方案，从而可以通过 EPR 检测肺损伤中的细胞和线粒体超氧化物。通过 EPR 测量超氧化物可以区分有肺损伤和无肺损伤的小鼠，以及具有不同疾病易感性的小鼠品系。我们希望这些方案能捕捉到超氧化物的实时生成情况，并将肺部 EPR 成像评估作为一种潜在的临床工具，用于根据氧化还原状态对 ARDS 患者进行亚型分型。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

Molecular Imaging and Biology 医学-核医学

CiteScore

6.90

自引率

3.20%

发文量

审稿时长

3 months

期刊介绍： Molecular Imaging and Biology (MIB) invites original contributions (research articles, review articles, commentaries, etc.) on the utilization of molecular imaging (i.e., nuclear imaging, optical imaging, autoradiography and pathology, MRI, MPI, ultrasound imaging, radiomics/genomics etc.) to investigate questions related to biology and health. The objective of MIB is to provide a forum to the discovery of molecular mechanisms of disease through the use of imaging techniques. We aim to investigate the biological nature of disease in patients and establish new molecular imaging diagnostic and therapy procedures. Some areas that are covered are: Preclinical and clinical imaging of macromolecular targets (e.g., genes, receptors, enzymes) involved in significant biological processes. The design, characterization, and study of new molecular imaging probes and contrast agents for the functional interrogation of macromolecular targets. Development and evaluation of imaging systems including instrumentation, image reconstruction algorithms, image analysis, and display. Development of molecular assay approaches leading to quantification of the biological information obtained in molecular imaging. Study of in vivo animal models of disease for the development of new molecular diagnostics and therapeutics. Extension of in vitro and in vivo discoveries using disease models, into well designed clinical research investigations. Clinical molecular imaging involving clinical investigations, clinical trials and medical management or cost-effectiveness studies.