Role of heparin binding growth factors in vascular leakage and fatal bleeding

肝素结合生长因子在血管渗漏和致命性出血中的作用

• 批准号: 7936131
• 负责人: PATRICIO E RAY
• 金额: $ 43万
• 依托单位: CHILDREN'S RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-21 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7936131
• 关键词: 2-tyrosine; Adenovirus Vector; Angiopoietin-1; Avian Leukosis Virus; Binding Proteins; Biological Assay; Biological Markers; Birds; Blood Circulation; Blood Vessels; Bypass; Capillary Permeability; Cardiac; Cardiopulmonary Bypass; Chickens; Child; Clinical; Coagulants; Critically ill children; Data; Development; Disease; Dose; Embryo; Endothelial Cells; Event; Extracorporeal Membrane Oxygenation; Extravasation; Fibroblast Growth Factor; Fibroblast Growth Factor 2; Fibroblast Growth Factor Receptor 2; Growth Factor; Harvest; Hemodialysis; Hemorrhage; Hemorrhagic Disorders; Heparin; Heparin Binding; Heparin Binding Growth Factor; Heparinoids; Herpesviridae; Human; Knowledge; Ligands; Mus; Neoplasms in Vascular Tissue; Oncogenes; Outcome; Pathogenesis; Pathway interactions; Pentosan Polysulfate; Permeability; Pharmaceutical Preparations; Plasma; Play; Polyomavirus; Procedures; Process; Relative (related person); Retroviral Vector; Rho-associated kinase; Risk; Role; Sampling; Signal Pathway; Testing; Tetanus Helper Peptide; Transgenes; Transgenic Mice; Tyrosine Kinase Inhibitor; Urine; Vascular Endothelial Growth Factor Receptor-2; Vascular Endothelial Growth Factors; Viral Tumor Antigens; Virus Receptors; Wild Type Mouse; Work; base; cell behavior; high risk; improved; in vivo; kinase inhibitor; novel; public health relevance; research study; sialosyl-T antigen; vector

DESCRIPTION (provided by applicant): Very little is known about the role that circulating heparin binding angiogenic growth factors play in the pathogenesis of bleeding disorders induced by heparin-like drugs or heparinoids in critically ill children. We have found a new role for Fibroblast Growth Factor -2 (FGF-2), an angiogenic heparin binding growth factor (HBGF), in the pathogenesis of bleeding disorders induced by heparin and pentosan polysulfate, an heparin-like drug. Others have found that an FGF binding protein (BP-1) that enhances the activity of FGF-2 induces vascular leakage and bleeding in the chicken CAM assay. Based on these findings, we hypothesize that FGF-2, alone or in combination with BP-1 or VEGF-A, enhances the capillary permeability changes induced by heparin-like drugs and increases their risk of causing lethal hemorrhages. This hypothesis will be tested in three specific aims: Aim 1 will define first the basic signaling pathways by which FGF-2, BP-1, VEGF-A, and plasma urine samples harvested from critically ill children, alone or in combination with heparin, increase the permeability of cultured human microvascular endothelial cells (HMVEC), and determine the relative contribution of the VEGFR-2, Tie-2, Src, and Rho kinase pathways in this process. Second, plasma/urine samples harvested from critically ill children with high plasma levels of FGF-2 treated with or without heparin, will be used to explore similar pathways and develop an HMVEC-permeability bioassay to discover new biomarkers to identify children at risk of bleeding when treated with heparin. In aim 2 tet-inducible Tg mice will be used to determine the relative contribution of BP-1 in the pathogenesis of these bleeding disorders. In addition, endothelial cells from TVA-Tg mice will be transduced with angiogenic retroviral vectors (RCAS), to enhance the release of FGF-2 and other HBGF into the circulation, and define their role in the pathogenesis of hemorrhages induced by heparinoids. Aim 3 will test the hypothesis that Angiopoietin-1 (Ang-1), an angiogenic anti-permeability growth factor, and the FGF-2 receptor tyrosine kinase inhibitor PD173074, will improve the clinical outcome of hemorrhages induced by heparinoids and FGF-2 in mice. The relative contribution and role of the Rho kinase pathway in this process will be explored as well. These experiments will generate new knowledge related to the role that HBGF released into the circulation of critically ill children play in the clinical outcome of hemorrhages induced by heparin, and test the novel concept that anti- permeability therapies will improve the clinical outcome of these children. PUBLIC HEALTH RELEVANCE: This proposal will test the hypothesis that heparin binding growth factors released into the circulation of critically ill children play a critical role in the development of vascular leakage and lethal hemorrhages induced by heparin-like drugs. In addition, we will test the novel concept that anti-permeability therapies will improve the clinical outcome of lethal hemorrhages induced by heparin-like drugs in in wild type and transgenic mice. These findings could have wider clinical implications for the treatment of critically ill children subjected to extracorporeal procedures) that required the use of heparin (cardiac bypass, ECMO, and hemodialysis and young children with vascular tumors and other angiogenic diseases.

描述（由申请人提供）：关于循环肝素结合血管生成生长因子在危重儿童肝素样药物或类肝素引起的出血性疾病发病机制中的作用，我们知之甚少。我们发现了成纤维细胞生长因子-2 (FGF-2)，一种血管生成性肝素结合生长因子（HBGF），在肝素和聚硫酸戊聚糖（一种肝素样药物）诱导的出血性疾病的发病机制中的新作用。其他人发现，在鸡CAM实验中，FGF结合蛋白（BP-1）可以增强FGF-2的活性，从而诱导血管渗漏和出血。基于这些发现，我们假设FGF-2单独或与BP-1或VEGF-A联合可增强肝素样药物引起的毛细血管通透性变化，并增加其引起致死性出血的风险。这一假设将在三个特定目标中得到验证：目标1将首先定义从危重儿童身上采集的FGF-2、BP-1、VEGF-A和血浆尿样本，单独或联合肝素增加培养的人微血管内皮细胞（HMVEC）的通透性的基本信号通路，并确定VEGFR-2、tio2、Src和Rho激酶通路在这一过程中的相对贡献。其次，从接受或不接受肝素治疗的FGF-2血浆水平较高的危重儿童身上采集的血浆/尿液样本将用于探索类似的途径，并开发hvec渗透性生物测定法，以发现新的生物标志物，以识别接受肝素治疗时有出血风险的儿童。在目的2中，将使用诱导Tg小鼠来确定BP-1在这些出血性疾病发病机制中的相对贡献。此外，TVA-Tg小鼠的内皮细胞将被血管生成逆转录病毒载体（angiogenic逆转录病毒载体，RCAS）转导，以促进FGF-2和其他HBGF释放到循环中，并确定它们在肝素类诱导出血发病机制中的作用。Aim 3将验证血管生成抗渗透性生长因子ang1和FGF-2受体酪氨酸激酶抑制剂PD173074是否会改善肝素类和FGF-2诱导的小鼠出血的临床结果。Rho激酶途径在这一过程中的相对贡献和作用也将被探讨。这些实验将对释放到危重儿童循环中的HBGF在肝素所致出血的临床结局中所起的作用产生新的认识，并验证抗渗透性治疗将改善危重儿童临床结局的新概念。公共卫生相关性：该提案将验证释放到危重儿童循环中的肝素结合生长因子在肝素样药物诱导的血管渗漏和致死性出血的发展中起关键作用的假设。此外，我们将在野生型和转基因小鼠中测试抗渗透性疗法将改善肝素样药物诱导的致死性出血的临床结果的新概念。这些发现可能对需要使用肝素进行体外手术的危重儿童（心脏搭桥、ECMO和血液透析）以及患有血管肿瘤和其他血管生成疾病的幼儿的治疗具有更广泛的临床意义。