Tc-99m-Labled Cytokine Ligands and SPECT Inflammation Imaging of Ischemic Hearts

Tc-99m 标记的细胞因子配体和缺血心脏的 SPECT 炎症成像

• 批准号: 8079124
• 负责人: Zhonglin Liu
• 金额: $ 37.88万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8079124
• 关键词: Acute; Affinity; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Binding; Binding Proteins; Biological; Biological Assay; Biological Availability; Blood; Cardiac; Cardiovascular Diseases; Cardiovascular system; Cells; Cessation of life; Chimeric Proteins; Chronic; Cultured Cells; Cytokine Receptors; Data; Detection; Development; Diagnosis; Dilatation - action; Endothelium; Evolution; Exhibits; Failure; Fibroblasts; Functional disorder; Goals; Health; Heart; Heart failure; Human; IgG1; Image; Imaging Techniques; Immune response; In Vitro; Individual; Infarction; Inflammation; Inflammatory; Inflammatory Response; Injury; Interleukin-1; Interleukin-12; Interleukin-18; Interleukins; Intervention; Ischemia; Kinetics; Label; Laboratories; Lead; Left; Ligands; Measures; Mechanics; Metabolic; Methods; Modality; Modeling; Mus; Muscle Cells; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardial Reperfusion; Myocardium; Pathway interactions; Patients; Phase; Play; Property; Proteins; Radiolabeled; Radiopharmaceuticals; Rattus; Reaction; Recombinants; Regional Blood Flow; Reperfusion Injury; Reperfusion Therapy; Research; Role; Site; Stress; TNF gene; TNFRSF1A gene; TNFRSF1B gene; Technetium 99m; Techniques; Testing; Therapeutic; Time; Tissues; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; Up-Regulation; Ventricular; Ventricular Function; Ventricular Remodeling; anakinra; base; cytokine; improved; in vivo; innovation; insight; interleukin-18 binding protein; novel; radioligand; radiotracer; receptor; response; single photon emission computed tomography; tool; uptake

DESCRIPTION (provided by applicant): The intense inflammatory reaction following myocardial infarction and reperfusion has been implicated as a crucial factor, which ultimately leads to ventricular remodeling, heart failure, and cardiovascular death. Our long-term goal in our research is to develop highly effective SPECT imaging techniques to screen and diagnose patients with cardiovascular diseases and to guide their therapy. The objective of the present proposal toward this goal is to validate 99mTc-labeled bispecific cytokine ligands for SPECT imaging of inflammation induced by myocardial ischemia and reperfusion. Tumor necrosis factor (TNF-1), Interleukin-1 (IL-12), and Interleukin-18 (IL-18) are the most potent proinflammatory cytokines involved in the progress of ischemia- reperfusion and ventricular remodeling; their actions are regulated by soluble Type I/Type II TNF receptor (TNFR1/TNFR2), IL-1 receptor antagonist (IL-1ra), and IL-18 binding protein (IL-18bp), respectively. Two 99mTc-labeled recombinant bispecific proteins, [99mTc]IL-18bp-Fc-IL-1ra (99mTc-IL181) and [99mTc]TNFR2-Fc- IL-1ra (99mTc-TR21), each containing an amino-terminal segment that specifically binds to TNFR2 or IL-18, the Fc portion of human IgG1, and a carboxy-terminal segment with the sequence of IL-1ra, have been recently developed in our laboratory for high-affinity targeting inflammatory sites via dual-cytokine pathways. In this proposal, we aim to: (1) determine the inflammation-targeting properties and radiopharmaceutical kinetics of these two bispecific radioligands in comparison with 99mTc-labeled individual cytokine ligands, 99mTc-TNFR2- Fc, 99mTc-IL-1ra-Fc and 99mTc-IL-18bp-Fc, in vitro and in vivo; (2) correlate the uptake of 99mTc-IL181 and 99mTc-TR21 in acute infarct and non-infarct regions with inflammatory intensity and progression of myocardial injury; and (3) evaluate 99mTc-IL181 and 99mTc-ILTR21 uptake associated with progressive left ventricular dilatation and pathophysiological alterations during the chronic remodeling phase and anti-inflammation therapy. To ascertain the abilities of 99mTc-IL181 and 99mTc-TR21 for specific cytokine targeting, cell-based binding assay will be carried out in IL-1-dependent murine D10.G4.1 (D10) cells. We will explore the interactions of the cytokine radioligands and cardiac cells (myocytes, endothelium and fibroblasts). Furthermore, we propose using a state-of-the-art small-animal SPECT imager, FastSPECT II, to collect dynamic cardiac imaging data in ischemic-reperfused rat hearts. Kinetic data of inflammatory reactions will be correlated with the extent of myocardial infarct and changes in cardiac function. The proposed research will apply SPECT imaging techniques toward the development of an innovative specific tool for fundamentally understanding inflammatory response in ischemic-reperfused hearts, providing insights into the intergradations of the proinflammatory cytokine pathway at different phases, and timing therapeutic modalities to limit the intense inflammatory reaction before severe cardiac function failure. PUBLIC HEALTH RELEVANCE: Development of highly specific imaging agents for noninvasive detection of inflammation induced by heart attacks.

描述（由申请人提供）：心肌梗死和再灌注后的强烈炎症反应被认为是最终导致心室重塑、心力衰竭和心血管死亡的关键因素。我们研究的长期目标是开发高效的SPECT成像技术，以筛查和诊断心血管疾病患者并指导其治疗。本提案的目的是验证99 mTc标记的双特异性细胞因子配体用于心肌缺血和再灌注诱导的炎症的SPECT成像。肿瘤坏死因子（TNF-1）、白细胞介素-1（IL-12）和白细胞介素-18（IL-18）是参与缺血-再灌注和心室重构进展的最有效的促炎细胞因子;它们的作用分别由可溶性I型/II型TNF受体（TNFR 1/TNFR 2）、IL-1受体拮抗剂（IL-1 ra）和IL-18结合蛋白（IL-18 bp）调节。两种99 mTc标记的重组双特异性蛋白[99 mTc]IL-18 bp-Fc-IL-1 ra（99mTc-IL181）和[99mTc]TNFR2-Fc- IL-1ra（99 mTc-TR 21），每个含有特异性结合TNFR 2或IL-18的氨基末端区段、人IgG 1的Fc部分和具有IL-1 ra序列的羧基末端区段，最近在我们的实验室中开发了通过双细胞因子途径高亲和力靶向炎症部位。本研究的目的是：（1）通过与99 mTc标记的单个细胞因子配体（99 mTc-TNFR 2- Fc、99 mTc-IL-1 ra-Fc和99 mTc-IL-18 bp-Fc）的比较，确定这两种双特异性放射性配体的体内外炎症靶向性和放射性药物动力学;（2）急性梗死和非梗死区99 mTc-IL 181和99 mTc-TR 21的摄取与炎症强度和心肌损伤进展相关;（3）评价99 mTc-IL 181和99 mTc-ILTR 21摄取与慢性重构期进行性左心室扩张和病理生理改变以及抗炎治疗的相关性。为了确定99 mTc-IL 181和99 mTc-TR 21对特异性细胞因子靶向的能力，将在IL-1依赖性鼠D10.G4.1（D10）细胞中进行基于细胞的结合测定。我们将探讨细胞因子放射性配体和心脏细胞（肌细胞，内皮细胞和成纤维细胞）的相互作用。此外，我们建议使用最先进的小动物SPECT成像仪，FastSPECT II，收集缺血再灌注大鼠心脏的动态心脏成像数据。炎症反应的动力学数据将与心肌梗死的程度和心功能的变化相关。拟议的研究将应用SPECT成像技术开发一种创新的特异性工具，用于从根本上了解缺血再灌注心脏的炎症反应，提供对不同阶段促炎细胞因子途径的相互作用的见解，并定时治疗方式，以限制严重心功能衰竭之前的强烈炎症反应。公共卫生相关性：开发高度特异性的成像剂，用于无创检测心脏病发作引起的炎症。