Utilization of Collagen Remodeling Pathways to Control Gene Delivery

1159466/Sullivan智能优点：拟议项目的总体目标是开发DNA连接的胶原水凝胶，其通过成纤维细胞的重塑将刺激DNA释放和高效的基因传递。最终，这些新型支架将被用来触发强大(且低成本)的原位生长因子(GFS)表达，以实现慢性伤口和其他受损组织的愈合。在所提出的方法中，通过将DNA结合多肽(DBPs)和胶原模拟多肽(CMPS)与先前确定的具有基因转移(DBPs)和胶原修饰(CMPS)能力的新的DNA结合多肽(DBPs)和胶原模拟多肽(CMPS)连接起来，用质粒DNA修饰胶原支架。这项拟议的工作将检验以下假设：(1)涉及胶原降解[由基质金属蛋白酶(MMPs)]和胶原内吞[由α2β1整合素和小窝蛋白-1]的天然胶原重塑过程将触发胶原连接的DNA“复合体”的释放和内吞；(2)胶原连接将通过涉及小窝蛋白-1的有效基因传递途径启动DNA运输。拟议目标的完成将从根本上为理解和指导基因传递提供新的策略，并从长远来看，将在组织修复中培育新的材料和治疗策略。广泛的影响：鉴于靶向GFs的丰富和胶原基材料的广泛使用，拟议的研究将不仅促进胶原介导的基因传递在慢性伤口修复中的应用，而且还将在植入功能和再生医学等其他多种应用中得到推广。拟议的努力还旨在通过建立在UD现有的本科生研究和推广基础设施以及Kiick和Sullivan以前的教育活动基础上，为本科生、高中实习生和代表性不足的群体提供新的研究和职业发展机会，包括行业指导。

1159466/ SullivanIntellectual Merit: The overall objective of the proposed project is to develop DNA-linked collagen hydrogels whose remodeling by fibroblast cells will stimulate DNA release and efficient gene delivery. Ultimately, these novel scaffolds will be used to trigger robust (and low cost) in situ expression of growth factors (GFs) to enable healing of chronic wounds and other damaged tissues. In the proposed approach, collagen scaffolds will be modified with plasmid DNAs by newly linking DNA-binding peptides (DBPs) and collagen mimetic peptides (CMPs) with previously established capacities for gene transfer (DBPs) and collagen modification (CMPs). The proposed work will test the following hypotheses: (1) native collagen remodeling processes involving collagen degradation [by matrix metalloproteinases (MMPs)] and collagen endocytosis [by alpha2 beta1 integrins and caveolin-1] will trigger the release and endocytosis of collagen-linked DNA "polyplexes"; (2) collagen linkage will initiate DNA trafficking through efficient gene delivery routes involving caveolin-1. Completion of the proposed aims should provide fundamentally new strategies to understand and direct gene delivery, and in the long term, will foster new materials and therapeutic strategies in tissue repair.Broader Impacts: Given the wealth of target GFs and the widespread use of collagen-based materials, the proposed studies will advance the use of collagen-mediated gene delivery not only in chronic wound repair, but also in multiple other applications such as implant functionalization and regenerative medicine. The proposed efforts are also designed to provide new research and career development opportunities, including industrial mentorship, for undergraduates, high school interns, and underrepresented groups, by building on existing undergraduate research and outreach infrastructure at UD, as well as on previous educational activities of Kiick and Sullivan.