Molecular Heparan Sulfate Delivery in Wound Regeneration

描述（申请人提供）：组织工程、移植手术、再生和组织重塑正在发展具有巨大潜在医疗价值的治疗方法。这些治疗方式都不是完全有效或可预测的。糖胺聚糖修饰的蛋白聚糖，如perlecan，曾经被认为主要是作为细胞外基质的结构成分，现在被关注于它们在组织和细胞调节中的作用，特别是血管生成和伤口愈合。Perlecan结构域1在体内与硫酸肝素一起糖基化，因此有可能影响伤口愈合的几个方面，并且是诱导调节伤口愈合的极好候选者。这个项目的前提是假设通过perlecan结构域1传递硫酸肝素将改善伤口愈合。Agenta Biotechnologies拥有与perlecan在伤口愈合和再生中的核酸载体表达相关的知识产权。在第一期工程中，我们将完成两个与骨再生有关的目标。为了建立可复制的体外参数，我们将测量表达perlecan结构域1的复制缺陷腺病毒向内皮细胞和成骨细胞的传递。为了在体内建立原理证明，我们将在小鼠骨缺损模型中评估腺病毒颗粒的递送，perlecan结构域1和硫酸肝素的表达，以及与血管发生和新骨形成相关的伤口愈合参数。在每个目标中，粒子传递，通过一种常见的骨移植替代物，羟基磷灰石晶体，将被评估。该项目第一阶段的成果将适用于许多伤口愈合学科，但在该项目第二阶段将首先针对骨再生的改善，届时将研究替代骨移植物替代品、屏障膜和外科应用，并将进行更广泛的装载、传递和表达的生化分析。分子硫酸肝素递送有可能改善骨移植和骨移植替代品的有效性，这是该技术平台和拟议项目的最终目标。

DESCRIPTION (provided by applicant): Tissue engineering, grafting procedures, regeneration, and tissue remodeling are developing therapies with great potential medical value. None of these therapeutic modalities are yet fully effective or predictable. Glycosaminoglycan-decorated proteoglycans such as perlecan, once thought to primarily serve as structural components of extracellular matrix, are now being focused on for their role in tissue and cell regulation, particularly angiogenesis and wound healing. Perlecan's domain 1 becomes glycosylated with heparan sulfate in vivo so has the potential to effect several aspects of wound healing and is an excellent candidate for induction in an effort to modulate wound healing. This project is premised on the hypothesis that delivery of heparan sulfate via the perlecan domain 1 will improve wound healing. Agenta Biotechnologies owns IP related to nucleic acid vector presentation of perlecan in wound healing and regeneration. In phase 1 of the project we will complete two aims related to osseous regeneration. To establish replicable in vitro parameters, we will measure delivery of replication-defective adenovirus expressing perlecan domain 1 to endothelial cells and osteoblasts. To establish proof of principle in vivo, we will assess delivery of the adenovirus particles, perlecan domain 1 and heparan sulfate expression, and wound healing parameters relating to vasculogenesis and new bone formation in a mouse osseous defect model. In each aim, particle delivery, via a common bone graft substitute, hydroxyapatite crystals, will be assessed. Outcomes of this phase 1 project will be applicable to many wound healing disciplines but will be targeted first to improvement of osseous regeneration in phase 2 of this project where alternative bone graft substitutes, barrier membranes, and surgical applications will be investigated, and a more extensive biochemical analysis of loading, delivery, and expression will be performed. Molecular heparan sulfate delivery has the potential to improve osseous grafting and the usefulness of bone graft substitutes, which is the ultimate goal of this technology platform and proposed project.