BOEC in Biology

BOEC 生物学

• 批准号: 6609692
• 负责人: ROBERT P HEBBEL
• 金额: $ 29.7万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-08 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6609692
• 关键词: NOD mouse; SCID mouse; apoptosis; bone marrow; cell biology; clinical research; coagulation factor VIII; gene expression; gene therapy; hematopoietic stem cells; hematopoietic tissue transplantation; histamine; histogenesis; human tissue; integrins; interleukin 6; intravenous administration; monoclonal antibody; polymerase chain reaction; selectins; spleen; stem cell transplantation; tissue /cell culture; transfection /expression vector; vascular cell adhesion molecule; vascular endothelial growth factors; vascular endothelium

DESCRIPTION (provided by applicant): Our preliminary data indicate that molecularly engineered BOEC (blood outgrowth endothelial cells) can be used for effective gene therapy of hemophilia A. The present project will examine several aspects of BOEC biology that are directly relevant to this potential therapeutic use of BOEC. Three Specific Aims will utilize the model of human BOEC (either unmanipulated or engineered to express human factor VIII) given to NOD. SCID mice. In Aim 1 we will identify the marrow "homing" behavior of BOEC. We will determine where BOEC go in the short- and long-terms when given intravenously. We will identify mechanisms underlying BOEC seeding of marrow and spleen. We will determine if BOEC seeding of marrow/spleen can be increase pharmacologically (with histamine, or beta-1-integrin activating antibody, or VEGF, or 1L6 plus stem cell factor). We will determine if the apparently low seeding frequency of BOEC in marrow is a stochastic or pre-programmed characteristic of establishing a BOEC graft. In Aim 2 we will examine certain issues regarding BOEC graft expansion and longevity in vivo. We will confirm our preliminary impression that BOEC expand substantially in vivo after being administered intravenously. We will determine if this in vivo expansion is predicatble and is dependent on the prior passaging history of the BOEC. We will seek to confirm that silencing of our fVIlI transgene expression vector is not a problem in BOEC. in Aim 3 we expect to document that endothelial expansion in vivo after intravenous administration of BOEC is due to a true BOEC cell rather than a contaminating hematopoietic precursor cell. Each of these studies addresses an issue that is necessary to understand before BOEC technology can be applied for therapeutics. Aside from eventual therapeutics, these studies will define aspects of this unique cell type.

描述（由申请人提供）：我们的初步数据表明，分子工程BOEC（血液内皮细胞）可用于有效的a型血友病基因治疗。本项目将研究BOEC生物学的几个方面，这些方面与BOEC的潜在治疗用途直接相关。Three Specific Aims将利用给予NOD的人类BOEC模型（未经操纵或经过工程改造以表达人因子VIII）。SCID小鼠。在目标1中，我们将确定BOEC的骨髓“归巢”行为。我们将确定静脉注射BOEC的短期和长期去向。我们将确定骨髓和脾脏BOEC播种的机制。我们将确定骨髓/脾脏的BOEC种子是否可以增加药理学（使用组胺，或β -1整合素激活抗体，或VEGF，或1L6 +干细胞因子）。我们将确定骨髓中BOEC明显的低播种频率是建立BOEC移植物的随机或预编程特征。在目标2中，我们将研究一些关于BOEC移植物扩张和体内寿命的问题。我们将证实我们的初步印象，即BOEC在静脉注射后在体内大量扩张。我们将确定这种体内扩张是否可预测，是否依赖于BOEC先前的传代历史。我们将试图证实我们的fVIlI转基因表达载体的沉默在BOEC中不是一个问题。在目的3中，我们希望证明静脉注射BOEC后体内内皮细胞的扩张是由于真正的BOEC细胞而不是污染的造血前体细胞。在BOEC技术应用于治疗之前，每一项研究都解决了一个必要的问题。除了最终的治疗方法，这些研究将定义这种独特细胞类型的各个方面。