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

• 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
• 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

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，包括内膜和中膜的重塑。 通过基质沉积和中膜平滑肌细胞（VSMC）增殖并迁移到 新生内膜干扰素-γ（IFN-γ）在免疫介导的重塑中起着重要的作用 在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， 动脉病性、移植的人同种异体冠状血管鉴定出血管内皮生长因子 受体-2（VEGFR 2）和神经纤毛蛋白-2（Neuropilin-2），两者均为VEGF反应性受体，以及内皮/平滑肌 细胞源性神经纤毛蛋白样分子（ESDN），作为潜在的特异性GA标志物。由于已知的 VEGFR 2-α-v-β 3相互作用和α-v-β 3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α-v-β 3在GA期间是否受到调节。我们证明，α-v-β 3，ESDN和VEGFR 2都是高度上调和/或激活的早期事件在GA，符合细胞增殖动力学。VEGF可诱导Th 1细胞极化，激活α-v-β 3 整联蛋白α-v-β 3结合反过来可以增强VEGFR 2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的IFN-γ，有助于T细胞分化、IFN-γ加工和 类似DTH的反应。与此同时，VEGF通过其经典受体和神经纤毛蛋白样受体， 分子，调节VSMC整合素活化和α-v-β 3介导的细胞增殖/迁移，这是VSMC整合素活化的标志。 GA的特点。目前的具体建议包括：（1）确定VEGF是否有助于 在嵌合SCID中开发利用VEGF/VEGFR拮抗剂的产生IFN-γ的T细胞和GA 小鼠/人动脉移植物模型和同种异体T细胞-内皮细胞培养物中;（2）评估 α-v-β 3整合素活化/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-γ R和VEGFR信号传导的关系，使用SCID/人动脉移植 模型和α-v-β 3抗体介导的阻断，以及用活化的α-v-β 3靶向剂和microSPECT技术的体内GA成像;（3）确定GA中的ESDN诱导是IFN-γ介导的还是 VEGF依赖性，在嵌合小鼠模型中利用同种异体人PBMC或IFN-γ刺激，和 ESDN表达是否增强或终止最大GA应答;和（4）解决IFN-γ-γ-IFN-γ的表达。 心脏移植患者中的VEGFR/ESDN-alpha- v-beta3轴，与血管内超声相关-记录的GA 微血管定位的IFN-γ、IFN-γ调节的趋化因子、VEGF/VEGF受体、α-v-β 3和ESDN表达 在肌内膜活检标本中，对于分泌分子，在血清样品中。分子机制 将在体外研究目标1-3中体内鉴定的途径，并将研究与人GA的相关性。 在目标4中。拟议的研究将确立IFN-γ-α-v-β 3- VEGFR/ESDN轴在 GA的发展，从而确定敏感和实用的早期GA标记。