FIBRONECTIN AND CELL RECRUITMENT

描述：（改编自申请人的摘要）-中央 这一建议的假设是，特定的整合素是必需的， 皮肤成纤维细胞迁移和肉芽组织形成 伤口 在皮肤损伤和 肉芽组织积聚。 在此延迟期间，成纤维细胞增加 它们的临时基质受体和减少它们的胶原蛋白受体。 作者提出，富含纤维蛋白/纤维连接蛋白的临时基质是 部分诱导机制控制成纤维细胞表达 临时基质整合素是一种调节机制， 早期伤口愈合反应。 为了验证这一假设，目标1提出 以确定成纤维细胞整合素的表达和功能， 使用猪切除的皮肤伤口的肉芽组织发育 伤口范例以及人类皮肤伤口。 为了证明要求 为了在肉芽组织发育期间调节整联蛋白， 整合素表达将通过加入特异性的 抗整联蛋白抗体或环肽整联蛋白拮抗剂 或通过在植入后将外源性成纤维细胞引入伤口部位 体外调节其特异性整联蛋白。 在目标2中， 在暂时性免疫抑制剂的情况下控制人皮肤成纤维细胞整联蛋白 将在分子水平上确定基质与胶原基质 利用模拟皮肤伤口环境的体外系统。 这 目的将包括细胞因子和细胞外基质增强剂的研究 特异性整合素启动子中的元件，以及 反式激活因子刺激这些整联蛋白增强子元件。 目的3提出确定成纤维细胞需要哪些整合素 在临时基质原纤维上移动，以及成纤维细胞迁移 从胶原凝胶到纤维蛋白凝块，使用体外条件， 模拟早期创伤环境中的选择性事件。 为此目的， 整联蛋白将被抗体和环肽拮抗剂调节， 培养条件和分子生物学操作。 此外，本发明还提供了一种方法， 提出研究整合素如何通过以下方式促进细胞运动： 使用特异性抑制剂调节成纤维细胞第二信使途径 和反义寡核苷酸。

DESCRIPTION: (Adapted from the applicant's abstract) - The central hypothesis of this proposal is that specific integrins are required for fibroblast migration and granulation tissue organization in cutaneous wounds. A three day delay intervenes between cutaneous wounding and granulation tissue accumulation. During this delay fibroblasts increase their provisional matrix receptors and decrease their collagen receptors. The authors propose that the fibrin/fibronectin-rich provisional matrix is part of the induction mechanism controlling fibroblast expression of provisional matrix integrins, a regulatory mechanism that is essential for the early wound healing response. To test this hypothesis, Aim 1 proposes to determine the expression and function of fibroblasts integrins during granulation tissue development of cutaneous wounds using porcine excisional wound paradigms as well as human cutaneous wounds. To prove the requirement for the modulation of integrins during granulation tissue development, integrin expression will be experimentally regulated by addition of specific anti-integrin antibodies or cyclic peptide integrin antagonists to the wound site or by introducing exogenous fibroblasts to the wound site after in vitro modulation of their specific integrins. In Aim 2, the regulatory control of human skin fibroblast integrins in the context of provisional matrix vs collagen matrix will be determined at the molecular level utilizing in vitro systems that simulate cutaneous wound environments. This aim will include a study of cytokine and extracellular matrix enhancer elements in specific integrin promoters, and identification of transactivating factors that stimulate these integrin enhancer elements. Aim 3 proposes to determine which integrins are required for fibroblast movement over provisional matrix fibrils, and for fibroblast transmigration from collagen gels into fibrin clots, using in vitro conditions that simulate selective events in the early wound environment. To this end, integrins will be modulated by antibody and cyclic peptide antagonists, by culture conditions, and by molecular biological manipulations. In addition, it is proposed to investigate how integrins promote cell movement by modulation of fibroblast second messenger pathways using specific inhibitors and antisense oligonucleotides.