Molecular Heparan Sulfate Delivery in Wound Regeneration

描述（由申请人提供）：组织工程、移植手术、再生和组织重塑正在开发具有巨大潜在医学价值的治疗方法。这些治疗方式中没有一种是完全有效或可预测的。糖胺聚糖修饰的蛋白聚糖，如串珠素，曾经被认为主要作为细胞外基质的结构成分，现在正集中在他们的组织和细胞调节，特别是血管生成和伤口愈合的作用。串珠素结构域1在体内被硫酸乙酰肝素糖基化，因此具有影响伤口愈合的几个方面的潜力，并且是用于诱导以调节伤口愈合的极好候选者。该项目基于经由串珠素结构域1递送硫酸乙酰肝素将改善伤口愈合的假设。Alfreta Biotechnologies拥有与串珠素在伤口愈合和再生中的核酸载体呈递相关的知识产权。在项目的第一阶段，我们将完成与骨再生相关的两个目标。为了建立可复制的体外参数，我们将测量表达串珠素结构域1的复制缺陷型腺病毒向内皮细胞和成骨细胞的递送。为了在体内建立原理证据，我们将评估腺病毒颗粒的递送、串珠素结构域1和硫酸乙酰肝素的表达，以及小鼠骨缺损模型中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，将评估通过常见骨移植替代物羟基磷灰石晶体的颗粒输送。该第1阶段项目的结果将适用于许多伤口愈合学科，但将首先针对该项目第2阶段中骨再生的改善，其中将研究替代骨移植替代品、屏障膜和手术应用，并将对加载、递送和表达进行更广泛的生化分析。分子硫酸乙酰肝素递送具有改善骨移植和骨移植替代物的有用性的潜力，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.