Determining Host-microbiome guided oro-nasal fistula healing

确定宿主微生物组引导口鼻瘘愈合

• 批准号: 10545072
• 负责人: Steven L Goudy
• 金额: $ 19.52万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10545072
• 关键词: Address; Affect; Antibiotics; Bacteria; Biopsy; Body Surface; Cells; Child; Cleft Palate; Communication; Communities; Craniofacial Abnormalities; Data; Development; Eating; Enterococcus faecalis; Environmental Risk Factor; Event; Exogenous Factors; Fistula; Gene Expression; Genes; Germ-Free; Gut Mucosa; HIV Infections; Homeostasis; Human; Hydrogels; Immune; Impairment; Innovative Therapy; Intestines; Knowledge; Live Birth; Low Prevalence; Measures; Metagenomics; Microbe; Mission; Modality; Modeling; Molecular; Mucous Membrane; Mus; Nasal cavity; Nose; Operative Surgical Procedures; Oral; Oral Surgical Procedures; Oral cavity; Patients; Population; Preparation; Prions; Probiotics; Process; Public Health; Regulator Genes; Research; Science; Signal Pathway; Signal Transduction; Site; Skin; Speech; Structure; Surgeon; Surgical wound; Technology; Testing; Tissue Donors; Tissues; Transcript; United States National Institutes of Health; commensal bacteria; congenital anomaly; craniofacial; disorder risk; experimental study; feeding; gene network; healing; host microbiome; improved; infection risk; intestinal injury; microbial community; microbiome; microbiome analysis; microbiome composition; microbiota; mouse model; novel therapeutic intervention; novel therapeutics; oral commensal; oral microbiome; palate repair; prevent; probiotic supplementation; recruit; regenerative; regenerative therapy; repaired; transcriptome sequencing; wound; wound bed; wound care; wound healing

SUMMARY Cleft palate formation is one of the most common craniofacial anomalies in children and occurs in 1:1000 live births. Patients undergo multiple surgeries during their lifetime and, for many, the first surgery is the repair of their cleft palate. However, 60% of cleft palate patients suffer poor wound healing following surgery which leads to the development of an oro-nasal fistula (ONF) characterized by a continued communication between the oral and nasal cavities. The primary approach to repair an ONF is by using human donor tissue, which acts only as a physical barrier and carries the risks of disease associated with human donor tissue, such as prions or HIV infection. Therefore, there is an urgent need to develop new therapeutic strategies to improve wound healing after cleft palate surgical repair, and to reduce the cases of ONF. Wound healing has been studied extensively on the skin and particularly following intestinal injury. Intestinal wound healing efficiency is highly sensitive to environmental factors, especially the microbiome, where specific microbial community structures or supplementation with probiotic bacteria enhances the wound healing process. However, little is known about how the oral microbiome affects ONF wound healing. We directly address this gap in the knowledge and show preliminary data that creating an ONF in a murine model results in marked changes in the microbiome composition, with the complete disappearance of certain microbes, and blooms of other bacterial taxa. Based on this premise, in Aim 1, we will create ONF wounds in germ-free mice, or in conventionally raised mice treated with antibiotics, and assess healing rates and changes in the oral microbiome composition. We expect to show the influence of disrupting the oral microbiome on healing processes and ONF formation. In Aim 2, we will use a hydrogel to re-introduce commensal oral microbes lost following surgery to the site of the oral wound and assess healing rates. We expect to show that repletion of oral commensal bacteria to the wound site can promote wound healing processes. In addition, by assessing transcript enrichment by RNA-seq within ONFs in germ-free, antibiotic treated mice, and after the repletion of oral commensal bacteria, we expect to identify gene networks that function in healing within ONFs that are sensitive to the commensal oral microbiome. Thus, our overall hypothesis is that the oral commensal microbiome exerts positive modulatory influences on oral wound healing and may limit ONF formation. Discovering specific bacterial taxa within the oral cavity or wound tissue that positively influence healing will be essential information for the characterization of an eubiotic oral microbiome for wound healing. Our experiments will also generate critical information for clinicians about the prudent use of antibiotics after cleft palate surgery and the effects of depleting the oral microbiome during healing. We will also substantiate the approach of the repletion of the oral commensal bacteria using hydrogel technology as the vehicle to enhance oral wound healing. Together, these studies will yield critical data to inform new therapeutic modalities to lower the prevalence of ONFs following cleft palate surgery.

概括 腭裂形成是儿童中最常见的颅面异常之一，发生率为 1:1000 出生。患者一生中接受多次手术，对于许多人来说，第一次手术是修复 他们的腭裂。然而，60%的腭裂患者术后伤口愈合不良，导致 导致口鼻瘘（ONF）的发展，其特征是之间的持续沟通 口腔和鼻腔。修复ONF的主要方法是使用人类供体组织，其作用 仅作为物理屏障，并具有与人类供体组织相关的疾病风险，例如朊病毒 或艾滋病毒感染。因此，迫切需要开发新的治疗策略来改善伤口 腭裂手术修复后愈合，并减少 ONF 病例。伤口愈合已被研究 广泛作用于皮肤，特别是在肠道损伤后。肠道伤口愈合效率高 对环境因素敏感，尤其是微生物组，其中特定的微生物群落结构或 补充益生菌可以促进伤口愈合过程。然而，人们对此知之甚少 口腔微生物组如何影响 ONF 伤口愈合。我们直接解决这一知识差距并展示 初步数据表明，在小鼠模型中创建 ONF 会导致微生物组发生显着变化 组成，某些微生物完全消失，而其他细菌类群大量繁殖。基于 在此前提下，在目标 1 中，我们将在无菌小鼠或常规饲养的小鼠中制造 ONF 伤口 用抗生素治疗，并评估治愈率和口腔微生物组成的变化。我们期望 显示破坏口腔微生物群对愈合过程和 ONF 形成的影响。在目标 2 中，我们 将使用水凝胶将手术后丢失的共生口腔微生物重新引入口腔伤口部位 并评估治愈率。我们希望证明，口腔共生细菌补充到伤口部位可以 促进伤口愈合过程。此外，通过 RNA-seq 评估 ONF 内的转录物富集度 无菌、抗生素治疗的小鼠，在口腔共生细菌充满后，我们希望能够识别出 对共生口腔微生物组敏感的 ONF 内具有愈合功能的基因网络。因此， 我们的总体假设是口腔共生微生物组对口腔发挥积极的调节影响 伤口愈合并可能限制 ONF 的形成。发现口腔或伤口内的特定细菌分类群 对愈合产生积极影响的组织将是表征优生口腔的重要信息 伤口愈合的微生物组。我们的实验还将为临床医生提供有关以下方面的关键信息： 腭裂手术后谨慎使用抗生素以及期间消耗口腔微生物组的影响 康复。我们还将证实使用水凝胶补充口腔共生细菌的方法 技术作为增强口腔伤口愈合的载体。这些研究将共同​​产生关键数据 告知新的治疗方式，以降低腭裂手术后 ONF 的患病率。