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: 10373683
• 负责人: Steven L Goudy
• 金额: $ 23.44万
• 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/10373683
• 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; 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; base; 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; rate of change; recruit; regenerative; regenerative therapy; repaired; transcriptome sequencing; wound; wound bed; 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.

总结 腭裂是儿童最常见的颅面畸形之一，发生率为1：1000。 出生患者在其一生中经历多次手术，对许多人来说，第一次手术是修复 他们的腭裂然而，60%的腭裂患者在手术后伤口愈合不良， 导致口鼻瘘（ONF）的发展，其特征是 口腔和鼻腔。修复ONF的主要方法是使用人类供体组织， 仅作为物理屏障，并具有与人类供体组织相关的疾病风险，如朊病毒 或者艾滋病病毒感染因此，迫切需要开发新的治疗策略来改善创伤 愈合腭裂手术修复后，并减少ONF的情况下，伤口愈合进行了研究 广泛地在皮肤上，特别是在肠损伤之后。肠创伤愈合效率高 对环境因素敏感，特别是微生物组，其中特定的微生物群落结构或 补充益生菌可增强伤口愈合过程。然而，人们对 口腔微生物组如何影响ONF伤口愈合。我们直接解决知识和显示的差距 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著变化 某些微生物完全消失，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创建ONF伤口 用抗生素治疗，并评估愈合率和口腔微生物组组成的变化。我们预计 以显示破坏口腔微生物组对愈合过程和ONF形成的影响。在目标2中， 将使用水凝胶将手术后丢失的口腔微生物重新引入口腔伤口部位 评估治愈率我们希望能证明口腔粘膜细菌在伤口部位的繁殖， 促进伤口愈合过程。此外，通过评估ONFs中RNA-seq的转录物富集， 无菌，抗生素治疗的小鼠，并在口腔粘膜细菌的补充后，我们希望能够鉴定出 在对共生口腔微生物组敏感的ONF内发挥愈合功能的基因网络。因此，在本发明中， 我们的总体假设是，口腔粘膜微生物组对口腔粘膜的生长产生积极的调节作用。 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 对愈合有积极影响的组织将是表征益生菌口服制剂的基本信息。 微生物组对伤口愈合的影响我们的实验还将为临床医生提供关于 腭裂手术后谨慎使用抗生素以及在手术期间消耗口腔微生物组的影响 治愈我们还将证实使用水凝胶补充口腔粘膜细菌的方法 技术作为促进口腔伤口愈合的载体。总之，这些研究将产生关键数据， 提供新的治疗方式，以降低腭裂手术后ONF的患病率。