BOEC in Biology

描述（由申请人提供）：我们的初步数据表明，分子工程 BOEC（血液生长内皮细胞）可用于 A 型血友病的有效基因治疗。本项目将研究与 BOEC 的潜在治疗用途直接相关的 BOEC 生物学的几个方面。三个具体目标将利用给予 NOD 的人类 BOEC 模型（未经操作或经过改造以表达人类因子 VIII）。 SCID小鼠。在目标 1 中，我们将识别 BOEC 的骨髓“归巢”行为。我们将确定 BOEC 静脉注射时短期和长期的走向。我们将确定 BOEC 骨髓和脾脏播种的潜在机制。我们将确定骨髓/脾脏的 BOEC 接种是否可以在药理学上增加（使用组胺、或 β-1-整合素激活抗体、或 VEGF、或 1L6 加干细胞因子）。我们将确定骨髓中 BOEC 明显较低的播种频率是否是建立 BOEC 移植物的随机或预编程特征。在目标 2 中，我们将研究有关 BOEC 移植物扩张和体内寿命的某些问题。我们将确认我们的初步印象，即 BOEC 静脉内给药后在体内大幅扩增。我们将确定这种体内扩增是否可预测，并且取决于 BOEC 之前的传代历史。我们将设法确认我们的 fVIII 转基因表达载体的沉默在 BOEC 中不是问题。在目标 3 中，我们期望记录静脉内给予 BOEC 后体内内皮扩张是由于真正的 BOEC 细胞而不是污染的造血前体细胞。这些研究中的每一项都解决了 BOEC 技术应用于治疗之前必须了解的一个问题。除了最终的治疗方法之外，这些研究还将定义这种独特细胞类型的各个方面。

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.