An immunomodulatory approach to improve oral cavity wound healing

项目总结：腭裂患者（1：1000活产）均接受腭裂修复术;然而，60%的患者（1：1000活产） Velop并发症，包括口鼻瘘（ONF）形成。ONF的形成导致进食不良， 演讲腭裂修复术包括将腭粘膜瓣旋转到腭裂中，愈合受到物理因素的影响。 舌头的创伤和充满细菌的环境。目前，腭裂修复手术缺乏一个 再生重建选择，由于ONF需要多次重新修复，并增加成本和发病率。 外科医生使用同种异体真皮植入物，尽管有HIV/朊病毒传播的风险，但仅作为屏障， 减少ONF。我们的小组已经开发了一种新的小鼠ONF表型，并确定了一种独特的免疫抑制剂， 在促再生程序中指导ONF愈合过程的调节方法。FTY 720，一种活跃的 生物脂质，靶向鞘氨醇途径，优先吸引促再生单核细胞和巨噬细胞， 以改善ONF愈合。我们的长期目标是开发一种免疫生成方法来改善pal- 吃了伤口愈合。本申请的总体目标是确定ONF愈合的机制， 使用FTY 720纳米纤维以促再生的方式调节单核细胞和巨噬细胞的迁移。的 中心假设是炎性单核细胞和巨噬细胞是口服给药期间的主要反应， 空腔伤口愈合和FTY 720的递送优先吸引促再生单核细胞和mac， 从而改善伤口愈合。FTY 720通过鞘氨醇途径发出信号， 促再生单核细胞和巨噬细胞的募集。拟议研究的基本原理是 全面了解ONF形成期间的炎症反应机制， 提供旨在改变促再生单核细胞迁移的治疗靶点。以强有力的指导， 没有任何数据，包括一篇证明FTY 720控释片给药后ONF显著降低的论文， 该假设将通过追求两个具体目标进行测试：1）确定要求，时间和机制， 在ONF形成过程中炎症反应的内在贡献; 2）工程化一种可降解支架， 提供FTY 720的受控递送，以使用免疫生成性AP改善口腔伤口愈合， 接近在目标1中，我们将确定愈合ONF的关键炎性细胞募集，确定重新激活ONF的可能性。 获得单核细胞，并鉴定ONF后细胞因子和修复基因通路的改变 阵在目标2中，我们将测试FTY 720的最佳交付策略，FTY 720的定向能力， 用和不用单核细胞的促再生ONF伤口愈合，并确定FTY 720纳米 纤维对细胞因子和修复基因通路的影响。该研究在机理上具有创新性 描述了允许测试免疫调节策略的ONF鼠模型中的单核细胞迁移。 这项拟议中的研究意义重大，因为FTY 720是一种FDA批准的药物， 硬化症，可以快速跟踪到临床试验，以减少ONF的形成。

PROJECT SUMMARY: Cleft palate patients (1:1000 live births) all undergo palate repair; however, 60% de- velop complications including oronasal fistula (ONF) formation. ONF formation leads to poor feeding and speech. Cleft palate repair involves rotating palate mucosal flaps into the cleft and healing is affected by physi- cal trauma from the tongue and a bacteria laden environment. Currently, cleft palate repair surgeries lack a regenerative reconstructive option, require multiple re-repairs due to ONF and increase cost and morbidity. Surgeons use allograft dermal implants, despite the risk of HIV/prion transmission, to act solely as a barrier to reduce ONF. Our group has developed a novel murine phenocopy of ONF, and identified a unique immuno- modulatory approach to directing the ONF healing process in a pro-regenerative program. FTY720, an active biolipid, targets the sphingosine pathway to preferentially attract pro-regenerative monocytes and macrophag- es to improve ONF healing. Our long-term goal is to develop an immunoregenerative approach to improve pal- ate wound healing. The overall objective in this application is to determine the mechanism of ONF healing and use FTY720 nanofibers to tune the monocyte and macrophage migration in a pro-regenerative fashion. The central hypothesis is that inflammatory monocytes and macrophages are the predominant response during oral cavity wound healing and that delivery of FTY720 preferentially attracts pro-regenerative monocytes and mac- rophages leading to improved wound healing. FTY720 signals through the sphingosine pathway, increasing the recruitment of pro-regenerative monocytes and macrophages. The rationale for the proposed research is that a comprehensive and mechanistic understanding of the inflammatory response during ONF formation will provide therapeutic targets aimed at altering pro-regenerative monocyte migration. Guided by strong prelimi- nary data, including a paper demonstrating significant reduction in ONF following FTY720 nanofiber delivery, the hypothesis will be tested by pursuing two specific aims: 1) Determine the requirement, timing and mecha- nistic contributions of the inflammatory response during ONF formation; 2) Engineer a nanofiber scaffold to provide controlled delivery of FTY720 to improve oral cavity wound healing using an immunoregenerative ap- proach. In Aim 1 we will identify the critical inflammatory cell recruitment to healing ONF, determine the re- quirement of monocytes, and identify alteration in the cytokines and reparative gene pathways following ONF formation. In Aim 2, we will test the optimal FTY720 delivery strategy, the ability of FTY720 nanofiber to direct pro-regenerative ONF wound healing with and without monocytes, and determine the effects of FTY720 nano- fiber on the cytokines and reparative gene pathways. The proposed research is innovative by mechanistically describing monocyte migration in an ONF murine model that allows testing of immunomodulatory strategies. The proposed research is significant because it that FTY720, an FDA-approved drug approved to treat multiple sclerosis, could be fast tracked to clinical trials to reduce ONF formation.