In vivo properties of gene transfer vectors and delivery in cardiovascular system

具体目的：所有这些研究的基础假设是，需要充分了解基因转移载体的药代动力学和药效学特性，才能最终开发组织和细胞特异性载体靶向，以精确和有效地传递基因。因此，这些研究旨在描述基因转移到小鼠和非人类灵长类动物以及小鼠和灵长类动物胎儿中的命运和影响，并描述宿主对基因修改的反应。本研究的具体目的是：1.系统地将VSV-G伪型逆转录病毒和慢病毒载体导入小鼠和猕猴以及小鼠和非猕猴胎儿体内，研究VSV-G伪型逆转录病毒和慢病毒载体的分布、药理特性、转基因表达和细胞效应。2.研究初始载体与细胞表面受体相互作用的机制，特别是糖胺聚糖和选择素等主要附着分子的作用。3.确定宿主对载体及其表达的转基因的免疫应答的性质和程度；4.检测动脉粥样硬化病变细胞外低密度脂蛋白的特异性抗体在体内表达的效果。5.鉴定噬菌体展示多肽文库向小鼠和猕猴胎儿的递送，并使用体内噬菌体淘洗方法鉴定体内胎儿组织的多肽。

SPECIFIC AIMS: The hypothesis underlying all of these studies is that a full understanding of their pharmacokinetic and pharmacodynamic properties of gene transfer vectors will be required for the eventual development of tissue- and cell-specific vector targeting for precise and effective gene delivery. These studies are therefore intended to characterize the fate and effects of gene transfer into mice and a non- human primate and into the mouse and primate fetus and to characterize the host response to the genetic modification. The specific aims of the studies are: 1. to characterize the distribution and pharmacological properties, transgene expression and cellular effects of VSV-G-pseudotyped retrovirus and lentivirus vectors following systematic introduction into mice and macaques and into mouse and non-macaque fetuses. 2. to characterize mechanisms of initial vector interaction with cell surface receptors, with special attention to the role of primary attachment molecules such as glycosaminoglycans and selectins. 3. to determine the nature and extent of the host immune response to the vectors and their expressed transgenes; 4. to test the effects of in vivo expression of an antibody specific for the extra-cellular LDL of atherosclerotic lesions. 5. to characterize delivery of phage display peptide libraries to mouse and macaque fetuses and use in vivo phage panning methods to identify peptides that home to fetal tissues in vivo.