Gene delivery approaches to vascular graft elastogenesis

描述（由申请人提供）：产生组织工程血管（TEBV）的主要障碍之一是接种细胞不能产生足够量的交联弹性蛋白以实现正常动脉的机械特性。本提案的目的是优化成人平滑肌细胞增殖、排列和弹性发生的条件，以实现TEBV在培养物中的快速发育。基于蛋白聚糖多功能蛋白聚糖V3的剪接变体的过表达的基因工程策略在体外和体内均诱导成年哺乳动物细胞中的弹性发生，但减弱细胞增殖。该建议的目的是产生用于表达V3和其他弹性蛋白基因的可调控载体，以便可以控制弹性蛋白合成的幅度和时间。我们假设，该系统将能够优化细胞覆盖和弹性蛋白沉积在生物材料支架上。为了提供机械支持并促进对齐，将细胞接种到有凹槽的胶原膜上。对齐的细胞片将形成管状结构。将在生物反应器中对构建体进行径向应变，并评估细胞预排列和弹性发生的机械贡献。

DESCRIPTION (provided by applicant): Among the primary obstacles to generating tissue engineered blood vessels (TEBV) is the inability of seeded cells to produce sufficient amounts of cross-linked elastin to achieve the mechanical characteristics of normal arteries. The objective of this proposal is to optimize conditions for adult smooth muscle cell proliferation, alignment and elastogenesis to achieve rapid development of TEBV in culture. A genetic engineering strategy based on overexpression of a splice variant of the proteoglycan versican, V3, induces elastogenesis in adult mammalian cells both in vitro and in vivo, but attenuates cell proliferation. The objectives of this proposal are to generate regulatable vectors for expression of V3 and other elastogenic genes so that the magnitude and timing of elastin synthesis can be controlled. We hypothesize that this system will enable optimization of cell coverage and elastin deposition on biomaterial scaffolds. To provide mechanical support and promote alignment, cells will be seeded onto grooved collagen membranes. Sheets of aligned cells will be formed into tubular constructs. The constructs will be subjected to radial strain in a bioreactor and the mechanical contribution of cell pre-alignment and elastogenesis will be assessed.