Vasoprotective Actions of Autologous Cell Transfer

描述（由申请方提供）：本提案的目的是表征和增强急性血管损伤动物模型中培养修饰的单核细胞（CMMCs）和内皮生长细胞（EOCs）的血管保护作用。最近在嵌合体动物模型中的研究表明，骨髓来源的细胞参与对血管损伤的细胞反应。在适当条件下体外培养的循环单核细胞可呈现内皮表型。我们的目标是适应和修改这些自体细胞用于治疗目的。CMMC通过使用限定的条件朝向内皮表型培养外周血单核细胞（PBMC）来产生。在培养7天后，大多数早期CMMC表达CD 14（LPS受体和单核细胞标志物），尽管该异质群体还包括内皮生长细胞（EOC）的前体和脱落的成熟循环内皮细胞（两者均为CD 14-）。在稍后的时间点，这些培养物由EOC（CD 14-）的同质群体组成。我们实验室的工作已经证明了在直接血管损伤模型中局部递送的早期CMMCs和EOCs的有效血管保护作用。我们的第一个工作假设是，自体早期CMMCs和EOCs发挥血管保护作用，在受损的血管内皮依赖的方式。我们的第二个工作假设是，基因修饰将进一步增强EOCs的血管保护作用。我们的四个具体目标是：具体目标1：比较与早期CMMCs和EOCs的局部递送相关的血管保护作用。具体目标2：确定早期CMMC和EOC输送的血管保护作用的机制。具体目标3：优化来自遗传修饰的EOC的转基因表达并增强其局部血管保护作用。具体目标4：优化遗传修饰EOC的全身转基因表达，并确定其全身血管保护潜力。

DESCRIPTION (provided by applicant): The objective of this proposal is to characterize and enhance the vasoprotective effects of culture-modified mononuclear cells (CMMCs) and endothelial outgrowth cells (EOCs) in an animal model of acute vascular injury. Recent studies in chimeric animal models have demonstrated that bone marrow-derived cells participate in the cellular response to vascular injury. Circulating mononuclear cells, cultured in vitro under appropriate conditions, may assume an endothelial phenotype. Our goal is to adapt and modify these autologous cells for therapeutic purposes. CMMCs are generated by culturing peripheral blood mononuclear cells (PBMCs) towards an endothelial phenotype using defined conditions. Following 7 days in culture, the majority of early CMMCs express CD14 (the LPS receptor and a monocyte marker) although this heterogenous population also includes precursors to endothelial outgrowth cells (EOCs) and sloughed mature circulating endothelial cells (both CD14-). At later time points, these cultures consist of homogenous populations of EOCs (CD14-). Work in our laboratory has demonstrated potent vasoprotective effects of locally delivered early CMMCs and EOCs in a model of direct vascular injury. Our first working hypothesis is that autologous early CMMCs and EOCs exert vasoprotective actions in the injured vasculature in an endothelial dependent fashion. Our second working hypothesis is that genetic modification will further enhance the vasoprotective effects of EOCs. Our four specific aims are: Specific Aim 1: To compare the vasoprotective effects associated with local delivery of early CMMCs and EOCs. Specific Aim 2: To define the mechanisms responsible for the vasoprotective effects of early CMMC and EOC delivery. Specific Aim 3: To optimize transgene expression from genetically modified EOCs and to enhance their local vasoprotective actions. Specific Aim 4: To optimize systemic transgene expression from genetically modified EOCs and to determine their systemic vasoprotective potential.