IRF6 in the Inflammatory Phase of Cutaneous Wound Healing

描述（由申请人提供）：先天免疫系统在炎症的启动和适当的伤口愈合中至关重要。在调节免疫应答的分子中有干扰素调节因子（IRF）家族的转录因子。特别令人感兴趣的是IRF 6，它是该家族的独特成员，也是伤口愈合的转录调节因子。事实上，患有货车德沃德综合征（VWS）（一种由IRF 6单倍不足引起的常染色体显性口面裂综合征）的患者比患有非综合征形式的口面裂的患者更可能在矫正手术后发生伤口并发症。Irf 6的缺失导致小鼠的颅面、肢体和表皮异常，导致新生儿死亡。我们最近发现，在肉芽组织中存在强Irf 6信号， 组织切除伤口，特别是在巨噬细胞和中性粒细胞。初步数据显示，与对照组相比，VWS患者的中性粒细胞体外趋化性受损。此外，在一个独特的鼠模型中，中性粒细胞和巨噬细胞中Irf 6的条件性去除导致简单炎症模型中细胞流入减少三倍。最后，皮肤伤口愈合严重受损七天后受伤。虽然其他IRF家族成员在先天免疫中的功能已得到充分描述，但对IRF 6在先天免疫细胞中的功能以及它如何影响皮肤伤口愈合的炎症阶段一无所知。 该提议的中心假设是，在伤口愈合的背景下，Irf 6是体外和体内适当的巨噬细胞和中性粒细胞功能所必需的。在具体目标#1中，我们将使用Luminex测定和迁移测定的活体成像来检验Irf 6调节巨噬细胞和中性粒细胞的分泌和迁移的假设。在具体目标#2中，我们将使用我们的Irf 6条件性敲除小鼠与溶菌酶驱动的Cre重组酶小鼠模型杂交，直接测试先天免疫细胞中Irf 6的表达对于伤口愈合的适当炎症阶段是必需的这一假设。 该提议是创新的，因为它将最近发现的Irf 6在表皮形态发生和生物学中的作用扩展到先天免疫和皮肤伤口愈合-从临床角度来看，这是一个非常重要的过程，也是健康相关成本上升的主要原因。这些研究完成后，我们将更了解Irf 6对 皮肤伤口愈合过程中的炎症过程。由于持续性炎症是导致慢性伤口、慢性阻塞性肺病的一种现象，并且也与癌症有关，因此将获得关于这些其他常见健康问题的潜在病理生理机制的新信息。 公共卫生相关性：与公共卫生的相关性本提案将评估干扰素调节因子6在皮肤伤口愈合背景下炎症中的作用，皮肤伤口愈合是健康相关成本上升的主要原因。我们希望更好地了解身体如何在受伤后关闭漏洞并形成接缝。这项工作可以应用于任何炎症性疾病，包括糖尿病，自身免疫性疾病，慢性伤口和癌症。

DESCRIPTION (provided by applicant): The innate immune system is of critical importance in the initiation of the inflammation and proper wound healing. Amongst molecules modulating the immune response are the transcription factors of the interferon regulatory factor (IRF) family. Of particular interest is IRF6, a unique member of this family and a transcriptional regulator of wound healing. Indeed, patients with Van der Woude syndrome (VWS), an autosomal dominant orofacial clefting syndrome caused by IRF6 haploinsufficiency, are more likely to have wound complications following corrective surgery than patients with a non-syndromic form of orofacial clefting. Loss of Irf6 results in craniofacial, limb and epidermal anomalies in the mouse, leading to neonatal mortality. We recently identified the presence of strong Irf6 signal in the granulation tissue of excisional wounds, particularly in macrophages and neutrophils. Preliminary data show that neutrophils from patients with VWS have impaired chemotaxis in vitro compared to controls. Furthermore, conditional removal of Irf6 in neutrophils and macrophages in a unique murine model results in a three fold reduction of cellular influx in a simple model of inflammation. Finally, cutaneous wound healing was severely impaired seven days post-wounding. Although the function of other IRF family members in innate immunity is well described, nothing is known about the function of IRF6 in innate immune cells and how it affects the inflammatory phase of cutaneous wound healing. The central hypothesis of this proposal is that Irf6 is required for proper macrophages and neutrophils function in vitro and in vivo in the context of wound healing. In specific aim #1, we will test the hypothesis that Irf6 regulates the secretion and migration of macrophages and neutrophils using luminex assay and live imaging of migration assays. In specific aim #2, we will directly test the hypothesis that expression of Irf6 in innate immune cells is necessary for the proper inflammatory phase of wound healing using our Irf6-conditional knockout mouse crossed with a lysosyme-driven Cre-recombinase mouse model. This proposal is innovative in that it extends the recently discovered role of Irf6 in epidermal morphogenesis and biology to innate immunity and cutaneous wound healing-a process that is highly significant from a clinical perspective, and a major cause of rising health-related costs. A the completion of these studies, we will have a better understanding of the contribution of Irf6 to the inflammatory process during cutaneous wound healing. Because continuous inflammation is a phenomenon leading to chronic wounds, chronic obstructive pulmonary disease, and is also implicated in cancer, new information will be obtained about a potential pathophysiological mechanism for these other common health concerns. PUBLIC HEALTH RELEVANCE: Relevance to Public Health This proposal will evaluate the role of Interferon Regulatory Factor 6 in inflammation in the context of cutaneous wound healing, a major cause of rising health-related costs. We expect to better understand how body closes holes and forms seams following injury. This work could have application to any inflammatory disease, including diabetes, autoimmune disorders, chronic wounds and cancer.