An IFN-y-Integrin-Growth Factor Axis in GA Biomarker Development

GA 生物标志物开发中的 IFN-γ-整合素生长因子轴

基本信息

批准号：
7491183
负责人：
JEFFREY R. BENDER
金额：
$ 36.73万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7491183
关键词：
Acute Address Adenoviruses Allogenic Allografting Animal Model Antibodies Arteries Arteriosclerosis Biological Markers Biopsy Specimen Blood Vessels CD4 Positive T Lymphocytes CXCL10 gene Cardiac Cell Differentiation process Cell Proliferation Chronic Classification Commit Coronary Vessels Deposition Detection Development Diagnostic Imaging Doctor of Medicine Doctor of Philosophy EGFR inhibition Early Diagnosis Endothelial Cells Event Failure Funding Future Goals Growth Factor Growth Factor Receptors Hepatocyte Human Hypoxia Image Immune Immunosuppression In Vitro Integrin alpha Chains Integrin beta3 Integrins Interferon Type II Invasive Jordan Kinetics Knowledge Localized Medial Mediating Methods Modeling Molecular Myocardial Ischemia Neuropilin-2 Neuropilins Organ Transplantation Pathway interactions Peripheral Blood Mononuclear Cell Play Polymerase Chain Reaction Production RNA Interference Reagent Recombinants Research Personnel Rest Role SCID Mice Sampling Serum Shapes Signal Transduction Smooth Muscle Myocytes Stimulus T-Lymphocyte Technology Th1 Cells Therapeutic immunosuppression Time Tissues Transplant Recipients Transplantation Ultrasonography Vascular Endothelial Growth Factor Receptor Vascular Endothelial Growth Factor Receptor-2 Vascular Endothelial Growth Factors Vascular remodeling Vision base cDNA Arrays chemokine heart allograft in vivo migration mouse model novel diagnostics programs receptor research study response

项目摘要

The hallmark of graft arteriosclerosis (GA) is vascular remodeling, which includes intimal and medial expansion by matrix deposition and medial smooth muscle cell (VSMC) proliferation with migration into the neointima. Interferon-gamma (IFN-y) plays a causative and key role in immune-mediated remodeling responses typical of those observed in GA. Our cDNA array and immunohistochemical analyses of arteriopathic, explanted human allograft coronary vessels identified vascular endothelial growth factor receptor-2 (VEGFR2) and Neuropilin-2, both VEGF-responsive receptors, and endothelial/smooth muscle cell-derived neuropilin-like molecule (ESDN), as potential specific GA markers. Because of the known VEGFR2-alpha-v--beta3 interactions and the role of alpha-v-beta3 in VSMC migration/proliferation, we utilized the chimeric human arterial transplant/SCID mouse model of GA to address whether alpha-v-beta3 is modulated during GA. We demonstrated that alpha-v-beta3, ESDN and VEGFR2 are all highly upregulated and/or activated as early events in GA, consistent with cell proliferation kinetics. VEGF can induce Th1 cell polarization and activate alpha-v-beta3 integrin. alpha-v-beta3 engagement can, in turn, enhance VEGFR2-mediated signaling. We hypothesize that VEGF, induced by tissue hypoxia and/or alloreactivity, contributes to T cell differentiation, IFN-y elaboration and consequent DTH-like responses. In parallel, VEGF, through its classical receptors and neuropilin-like molecules, modulates VSMC integrin activation and alpha-v-beta3-mediated cell proliferation/migration, a hallmark feature of GA. Specific proposals now include to: (1) determine whether VEGF contributes to the development of IFN-y- producing T cells and GA utilizing VEGF/VEGFR antagonists in the chimeric SCID mouse/human arterial graft model, and in allogeneic T cell-endothelial cell cultures; (2) assess the causative role of alpha-v-beta3 integrin activation/function in allogeneic peripheral blood mononuclear cell (PBMC)- and IFN-y-induced GA and its relationship to IFN-yR and VEGFR signaling, using the SCID/human arterial transplant model and alpha-v-beta3 antibody-mediated blockade, as well as in vivo GA imaging with an activated alpha-v-beta3-targeted agent and microSPECT technology; (3) determine whether ESDN induction in GA is IFN-y-mediated or VEGF-dependent, utilizing the allogeneic human PBMC or IFN-y stimuli in the chimeric mouse model, and whether ESDN expression enhances or terminates maximal GA responses; and (4) address the IFN-y- VEGFR/ESDN-alpha- v-beta3 axis in cardiac transplant patients, correlating intravascular ultrasound- documented GA with microvessel-localized IFN-y, IFN-y-regulated chemokines, VEGF/VEGFRs, alpha-v-beta3 and ESDN expression in endomyocardial biopsy specimens and, for secreted molecules, serum samples. Molecular mechanisms of the pathways identified in vivo in Aims 1-3 will be studied in vitro, and relevance to human GA will be addressed in Aim 4. The proposed studies will establish the validity of an IFN-y - alpha-v-beta3 - VEGFR/ESDN axis in the development of GA, thus defining sensitive and practical markers of early GA.

移植物动脉硬化（GA）的标志是血管重塑，包括内膜和内侧通过基质沉积和内侧平滑肌细胞 (VSMC) 增殖并迁移至新内膜。干扰素-γ (IFN-y) 在免疫介导的重塑中发挥着致病和关键作用 GA 中观察到的典型反应。我们的 cDNA 阵列和免疫组织化学分析动脉病，移植的人同种异体冠状动脉血管鉴定出血管内皮生长因子受体 2 (VEGFR2) 和 Neuropilin-2，均为 VEGF 响应受体，以及内皮/平滑肌细胞源性神经毡蛋白样分子 (ESDN)，作为潜在的特异性 GA 标记。因为众所周知 VEGFR2-alpha-v--beta3 相互作用以及 alpha-v-beta3 在 VSMC 迁移/增殖中的作用，我们利用 GA 的嵌合人动脉移植/SCID 小鼠模型来解决 GA 期间 alpha-v-beta3 是否受到调节。我们证明，α-v-β3、ESDN 和 VEGFR2 在 GA 早期事件中均高度上调和/或激活，与细胞增殖动力学一致。 VEGF 可诱导 Th1 细胞极化并激活 alpha-v-beta3 整合素。 alpha-v-beta3 的结合反过来又可以增强 VEGFR2 介导的信号传导。我们假设 VEGF，由组织缺氧和/或同种异体反应性诱导，有助于 T 细胞分化、IFN-y 的形成和随之而来的类似 DTH 的反应。与此同时，VEGF 通过其经典受体和神经毡蛋白样分子，调节 VSMC 整合素激活和 alpha-v-beta3 介导的细胞增殖/迁移，这是一个标志遗传算法的特征。现在的具体建议包括：（1）确定 VEGF 是否有助于在嵌合 SCID 中利用 VEGF/VEGFR 拮抗剂开发产生 IFN-γ 的 T 细胞和 GA 小鼠/人动脉移植模型以及同种异体 T 细胞-内皮细胞培养物； (2) 评估病因使用 SCID/人动脉移植物，研究 α-v-β3 整合素激活/功能在同种异体外周血单核细胞 (PBMC) 和 IFN-γ 诱导的 GA 中的作用及其与 IFN-γR 和 VEGFR 信号传导的关系模型和 α-V-β3 抗体介导的阻断，以及使用激活的 α-V-β3 靶向药物和 microSPECT 技术进行体内 GA 成像； (3) 确定 GA 中的 ESDN 诱导是否是 IFN-γ 介导的或 VEGF 依赖性，在嵌合小鼠模型中利用同种异体人类 PBMC 或 IFN-y 刺激，以及 ESDN 表达是否增强或终止最大 GA 响应； (4) 解决 IFN-y- 心脏移植患者中的 VEGFR/ESDN-α-v-β3 轴，与血管内超声记录的 GA 相关具有微血管定位的 IFN-γ、IFN-γ 调节的趋化因子、VEGF/VEGFR、α-v-β3 和 ESDN 表达心内膜心肌活检标本以及分泌分子血清样本。分子机制目标 1-3 中确定的体内途径将在体外进行研究，并且与人类 GA 的相关性将目标 4 中提出。拟议的研究将确定 IFN-y - alpha-v-beta3 - VEGFR/ESDN 轴在 GA 的发展，从而定义早期 GA 的敏感且实用的标记。