Fibronectin peptides for treatment of burns

描述（由申请人提供）：美国每年有超过100万例烧伤，导致70万例急诊就诊，45，000例住院，4，500例死亡。烧伤是一种动力性损伤，其特征是在最初几天内损伤逐渐扩大。部分皮层烧伤进展为全皮层烧伤与死亡率、伤口脓毒症和瘢痕形成的增加有关。旨在预防烧伤进展的方法一直是密集研究的焦点，但这些结果令人失望。事实上，烧伤治疗尚未解决的挑战是开发一种限制损伤、加速愈合和减少疤痕的治疗方法。发现新的方法来成功地解决这一挑战将是烧伤患者护理的一个重大进展。在过去的十年中，克拉克实验室对伤口中新组织的形成进行了几次重要的观察。这些体内研究刺激了一种新的体外细胞迁移模型的发展，如在早期愈合伤口中发现的那样。该测定还被用于筛选新的基质材料，因为它们能够支持细胞从胶原基质迁移到新基质中，如在伤口周围环境中所发现的。随后，发现3个纤连蛋白（FN）功能结构域对于成纤维细胞的运动是必要和充分的。从这一发现，分子间交联的透明质酸（HA）的3个FN结构域修饰的工程和证明，以提高成纤维细胞迁移在体外和猪皮肤伤口修复在体内。最近，在3个FN结构域中发现4个位点与血小板衍生生长因子-BB（PDGF-BB）结合。PDGF-BB在与这些FN生长因子（GF）结合结构域或肽结合时保持活性。重要的是，PDGF-BB作为FN缺陷细胞的存活因子的能力依赖于FN GF结合结构域或肽。由于烧伤创面可能缺乏FN通过热变性和酶降解，这些发现提出的可能性，FN GF结合域或肽可能是有用的烧伤治疗。在这里，我们建议创建一个局部制剂深部分厚度烧伤创面含有FN生长因子增强肽（GFEP）。为此，我们建议屏幕小（<15个氨基酸）FN肽结合PDGF-BB并促进FN缺失成纤维细胞存活和增殖的能力（AIM 1），测定FN PDGF结合肽与PDGF联合促进人成纤维细胞和微血管内皮细胞活化、增殖和迁移的能力（AIM 2），并在先前已验证的猪模型中测试FN PDGF结合肽与PDGF联合限制损伤、促进愈合和预防烧伤伤口瘢痕形成的能力（AIM 3）。美国每年有超过100万例烧伤，导致70万次急诊就诊，45，000次住院，4，500人死亡。旨在预防烧伤进展的方法一直是密集研究的焦点，但这些结果令人失望。在这里，我们建议创建一个局部制剂深部分厚度烧伤创面含有生长因子增强肽，最近发现在纽约州立大学斯托尼布鲁克，并有显着的能力，以促进组织培养的细胞存活和增殖。

DESCRIPTION (provided by applicant): There are more than 1 million burns each year in the US, resulting in 700,000 emergency department visits, 45,000 hospital admissions, and 4,500 deaths. Burns are dynamic injuries characterized by progressive extension of the injury over the first few days. Progression of partial thickness burns to full thickness injuries is associated with increases in mortality, wound sepsis, and scar formation. Methods aimed at preventing burn progression have been the focus of intense research but these results have been disappointing. In fact the unmet challenge of burn treatment is development of a therapy that limits injury, speeds healing and reduces scarring. Discovery of novel methods to successfully address this challenge would be a major advance in the care of patients with burns. Over the past decade the Clark laboratory has made several critical observations about new tissue formation in wounds. These in vivo investigations stimulated the development of a novel in vitro model of cell migration across matrix boundaries as would be found in early healing wounds. The assay has also been utilized to screen new matrix materials for their ability to support cell migration from a collagen matrix, as would be found in a periwound environment, into a neomatrix. Subsequently, 3 fibronectin (FN) functional domains were found to be necessary and sufficient for fibroblast movement. From this discovery, an intermolecular cross-linked hyaluronan (HA) decorated with the 3 FN domains was engineered and demonstrated to enhance fibroblast migration in vitro and in porcine cutaneous wound repair in vivo. Recently, 4 sites in 3 FN domains were found to bind platelet-derived growth factor-BB (PDGF-BB). PDGF-BB remained active while bound to these FN growth factor (GF)-binding domains or peptides. Importantly the ability of PDGF-BB to act as a survival factor for FN-deficient cells is dependent on FN GF-binding domains or peptides. Since burn wounds can be deficient in FN through thermal denaturation and enzymatic degradation, these findings raise the possibility that FN GF-binding domains or peptides may be useful in burn therapy. Here we propose to create a topical preparation for deep partial thickness burn wounds containing a FN growth factor-enhancing peptide (GFEP). To this end we propose to screen small (<15 amino acid) FN peptides for their ability to bind PDGF-BB and promote FN null fibroblast survival and proliferation (AIM 1), assay FN PDGF-binding peptides in conjunction with PDGF for their ability to promote activation, proliferation and migration of human fibroblasts and microvascular endothelial cells (AIM 2), and test FN PDGF-binding peptides in conjunction with PDGF for their capacity to limit injury, enhance healing and prevent scarring of burn wounds in a porcine model that has been previously validated (AIM 3). There are more than 1 million burns each year in the US, resulting in 700,000 emergency department visits, 45,000 hospital admissions, and 4,500 deaths. Methods aimed at preventing burn progression have been the focus of intense research but these results have been disappointing. Here we propose to create a topical preparation for deep partial thickness burn wounds containing a growth factor-enhancing peptide that was recently discovered at SUNY Stony Brook and has the remarkable ability to promote tissue cultured cell survival and proliferation.