Novel Diagnostics to Detect Lung Injury

检测肺损伤的新型诊断方法

• 批准号: 8803317
• 负责人: ELIZABETH R JACOBS
• 金额: --
• 依托单位: CLEMENT J. ZABLOCKI VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8803317
• 关键词: Adenine; Adopted; Affect; Amobarbital; Apoptosis; Biochemical; Bioenergetics; Brain; Bronchiolitis Obliterans; Caring; Cell Death; Cell Survival; Cells; Cessation of life; Chest; Chronic Obstructive Airway Disease; Clinical; Coenzymes; Complex; Crush Injury; Cyclosporine; Cystic Fibrosis; Data; Decision Making; Detection; Dinucleoside Phosphates; Discipline of Nuclear Medicine; Electrons; FADH2; FDA approved; Figs - dietary; Fluorescence; Functional disorder; Generations; Glutathione; Hand; Health; Heart; Histologic; Human; Image; Imaging Techniques; Immunosuppressive Agents; In Situ Nick-End Labeling; Injury; Intervention; Ischemia; Label; Learning; Left; Life; Lung; Lung Transplantation; Lung diseases; Measures; Mediating; Metabolic; Methods; Methylprednisolone; Mitochondria; Modeling; Monitor; Morbidity - disease rate; NADH; Necrosis; Nicotinamide adenine dinucleotide; Optics; Organ; Oxidation-Reduction; Oxidative Phosphorylation; Oximes; Pathology; Pharmaceutical Preparations; Pneumonia; Positioning Attribute; Process; Protocols documentation; Pulmonary Fibrosis; Pulmonary Hypertension; Rattus; Reactive Oxygen Species; Recovery; Reperfusion Injury; Reperfusion Therapy; Respiratory physiology; Risk Factors; Rodent; Role; Safety; Signal Transduction; Staging; Staining method; Stains; Steroids; Structure; Technetium 99m; Techniques; Testing; Time; Tissues; Translating; Transplantation; Transplanted Lung Complication; Vascular blood supply; Veterans; X-Ray Computed Tomography; caspase-3; clinical application; duramycin; fluorescence imaging; graft failure; imaging modality; imaging probe; improved; in vivo; indexing; inhibitor/antagonist; lung injury; lung ischemia; mitochondrial dysfunction; mitochondrial permeability transition pore; non-invasive imaging; novel; novel diagnostics; novel therapeutic intervention; operation; optical imaging; prevent; prognostic; prognostic value; sedative; single photon emission computed tomography; tool

DESCRIPTION (provided by applicant): Experts in lung transplantation agree that ischemia is a recognized risk factor for all postlung transplant problems. Better non-invasive methods to detect ischemia post transplant would provide a window to modify therapy, and thereby reduce morbidity of veterans from these complications. Ischemia-reperfusion (IR) lung damage is also encountered clinically in conditions beyond lung transplantation, including necrotizing pneumonias, or crush injury to the chest (e.g Farivar et al, 2005), all injuries that frequently affect veterans. Substantial evidence supports the role of mitochondrial dysfunction and mitochondrially derived reactive oxygen species (mtROS) in IR injuries to the brain and heart (Stowe & Camara, 2009). Our preliminary data support a critical role for mitochondria in a rat model unilateral lung IR injury, a model of lung transplant which eliminates immunological or drug induced pathology in order to focus exclusively on IR injury. We introduce 2 absolutely novel non-destructive methods to track apoptosis and the redox state in intact IR lung, methods that can be rapidly translated into the clinical field. The first is single photon emission computed tomography/computed tomography (SPECT/CT) imaging, using 99m Tc-duramycin (DU) and 99m Tc- hexamethylpropyleneamine oxime (HMPAO) to detect cell death/apoptosis and gluthathione/redox status respectively. The second is fluorescence optical imaging to follow the redox ratio (RR) of lung. We hypothesize that: (i) IR stimulates mitochondrial complex I dysfunction which leads to subsequent apoptosis and decreased cell survival as detected biochemically and histologically (ii) SPECT/CT and optical imaging can detect and quantify apoptosis or altered mitochondrial bioenergetics in vivo. Changes in the values of these imaging endpoints after IR will correlate with histological injury and some indices of mitochondrial dysfunction (iii) treatment with agents that prevent opening of mitochondrial permeability transition pore (mPTP) and those that are used clinically to prevent rejection will protect against lung injury as detected by SPECT/CT or optical imaging, and this protection will correlate to improved indices of mitochondrial function. We will test these hypotheses with three specific objectives/aims. (1) To use SPECT and nuclear medicine probes which target apoptosis/necrosis or oxidoreductive state to detect unilateral IR lung injury in vivo We will correlate serial SPECT data after IR injury to histological and biochemical evidence of apoptosis/necrosis one week after ischemia. (2) To employ optical imaging which detects the oxidoreductive state of tissue to follow IR lung injury in vivo. The mitochondrial metabolic coenzymes Nicotinamide Adenine Dinucleotide (reduced form is NADH) and Flavine Adenine Dinucleotide (FAD) are the primary electron carriers in oxidative phosphorylation. NADH and FAD (oxidized form of FADH2) are autofluorescent and can be monitored by optical techniques without the need for exogenous labels. (3) To investigate the potential of our novel imaging methods to detect protection from IR injury by agents which act on complex I and the mitochondrial permeability transition pore (mPTP) and that are used clinically in lung transplantation. With our novel means to detect apoptosis and redox injury and a survival model of lung IR injury, we are poised to examine the potential of these non-destructive methods to track ischemic Injury. Though not used routinely for detection of lung injury, SPECT imaging with HMPAO is already in clinical use with well established safety profiles. With translational evidence of prognostic value, these imaging modalities can be adapted readily to improve the care of veterans with IR lung damage. We are uniquely positioned to test novel therapeutic interventions to improve IR lung injury with prognostic and mechanistic information in hand.

描述（由申请人提供）： 肺移植专家一致认为，缺血是所有肺移植后问题的公认危险因素。更好的非侵入性方法来检测移植后缺血将为修改治疗提供一个窗口，从而降低退伍军人这些并发症的发病率。缺血再灌注 (IR) 肺损伤在临床上也出现在肺移植以外的情况下，包括坏死性肺炎或胸部挤压伤（例如 Farivar 等人，2005 年），这些损伤经常影响退伍军人。实质性证据 支持线粒体功能障碍和线粒体衍生的活性氧 (mtROS) 在大脑和心脏红外线损伤中的作用 (Stowe & Camara, 2009)。我们的初步数据支持线粒体在大鼠单侧肺 IR 损伤模型中的关键作用，这是一种肺移植模型，消除了免疫或药物诱导的病理，以便专门关注 IR 损伤。我们引入了两种绝对新颖的非破坏性方法来追踪完整 IR 肺中的细胞凋亡和氧化还原状态，这些方法可以快速转化为临床领域。第一种是单光子发射计算机断层扫描/计算机断层扫描（SPECT/CT）成像，使用99m Tc-耐久霉素（DU）和99m Tc-六甲基丙烯胺肟（HMPAO）分别检测细胞死亡/凋亡和谷胱甘肽/氧化还原状态。第二种是荧光光学成像，以追踪肺部的氧化还原比（RR）。我们假设：(i) IR 刺激线粒体复合物 I 功能障碍，导致随后的细胞凋亡和细胞存活率下降，如生化和组织学检测到的 (ii) SPECT/CT 和光学成像可以检测和量化细胞凋亡或体内线粒体生物能的改变。 IR 后这些成像终点值的变化将与组织学损伤和线粒体功能障碍的一些指标相关 (iii) 使用防止线粒体通透性转换孔 (mPTP) 打开的药物和临床上用于预防排斥反应的药物进行治疗将防止通过 SPECT/CT 或光学成像检测到的肺损伤，这种保护将与线粒体功能指数的改善相关。我们将通过三个具体目标来测试这些假设。 (1) 使用针对细胞凋亡/坏死或氧化还原状态的SPECT和核医学探针来检测体内单侧IR肺损伤我们将IR损伤后的系列SPECT数据与缺血后一周细胞凋亡/坏死的组织学和生化证据相关联。 (2) 采用光学成像技术检测组织的氧化还原状态，以跟踪体内红外肺损伤。线粒体代谢辅酶烟酰胺腺嘌呤二核苷酸（还原型NADH）和黄素腺嘌呤二核苷酸（FAD）是氧化磷酸化的主要电子载体。 NADH 和 FAD（FADH2 的氧化形式）具有自发荧光，可以通过光学技术进行监测，无需外源标记。 (3) 研究我们的新型成像方法检测作用于复合物 I 和线粒体通透性转换孔 (mPTP) 并临床用于肺移植的药物对 IR 损伤的保护作用的潜力。凭借我们检测细胞凋亡和氧化还原损伤的新方法以及肺IR损伤的生存模型，我们准备检验这些非破坏性方法追踪缺血性损伤的潜力。虽然不常规用于检测肺损伤，但 HMPAO 的 SPECT 成像已在临床使用，并具有良好的安全性。有了预后价值的转化证据，这些成像方式可以很容易地进行调整，以改善患有 IR 肺损伤的退伍军人的护理。我们拥有独特的优势，可以测试新的治疗干预措施，以改善 IR 肺损伤，并掌握预后和机制信息。