Vasoprotective Actions of Autologous Cell Transfer

描述（由申请人提供）：本提案的目的是表征和增强培养修饰的单核细胞（CMMCs）和内皮生长细胞（EOCs）在急性血管损伤动物模型中的血管保护作用。最近在嵌合动物模型中的研究表明，骨髓来源的细胞参与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能呈现内皮表型。我们的目标是适应和修改这些自体细胞用于治疗目的。cmmc是通过在特定条件下将外周血单个核细胞（PBMCs）培养成内皮表型而产生的。在培养7天后，大多数早期cmmc表达CD14 （LPS受体和单核细胞标记物），尽管这种异质群体也包括内皮生长细胞（EOCs）和脱落的成熟循环内皮细胞的前体（均为CD14-）。在后来的时间点，这些培养物由同质的EOCs （CD14-）群体组成。我们实验室的研究表明，在直接血管损伤模型中，局部递送的早期cmmc和EOCs具有强大的血管保护作用。我们的第一个工作假设是，自体早期cmmc和EOCs以内皮依赖的方式在受损的血管中发挥血管保护作用。我们的第二个假设是，基因改造将进一步增强EOCs的血管保护作用。我们的四个具体目标是：具体目标1：比较早期cmmc和EOCs局部递送的血管保护作用。具体目标2：明确CMMC和EOC早期交付血管保护作用的机制。特异性目的3：优化转基因EOCs的转基因表达，增强其局部血管保护作用。特异性目标4：优化转基因EOCs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.