Skin microbial-based mechanisms of accelerated wound healing

基于皮肤微生物的加速伤口愈合机制

• 批准号: 10464143
• 负责人: Ellen White
• 金额: $ 3.42万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10464143
• 关键词: Affect; Aftercare; Alcaligenes; Amputation; Bacteria; Biochemical; Biological Assay; Biology; Cell surface; Cells; Chronic; Clinical; Collection; Communication; Communities; Cytokine Signaling; Development; Diabetic Foot Ulcer; Doctor of Philosophy; Enterococcus faecalis; Epithelial; Failure; Fostering; Future; Gene Expression; Gene Expression Profiling; Genetic Determinism; Genetic Screening; Genomic Segment; Genomic approach; Genomics; Goals; Health Care Costs; Healthcare Systems; Immune response; Impaired healing; In Vitro; Inflammatory; Interleukin-6; Investigation; Knowledge; Lead; Mediating; Mentorship; Metagenomics; Microbial Genetics; Morbidity - disease rate; Mus; Necrosis; Osteomyelitis; Outcome; Pathway interactions; Pennsylvania; Phenotype; Physicians; Play; Production; Research; Research Personnel; Role; Sampling; Scientist; Series; Signal Transduction; Skin; Specimen; Staphylococcus aureus; Sterility; Stream; Techniques; Testing; Therapeutic; Training; Treatment Cost; Universities; Variant; Work; adverse outcome; base; cell behavior; chronic wound; commensal bacteria; comparative genomics; cytokine; diabetic ulcer; experimental study; genomic locus; gut microbiome; healing; host-microbe interactions; improved; in vivo; keratinocyte; microbial; microbial community; microbiome; microbiota; migration; mortality; mouse model; new therapeutic target; non-healing wounds; novel; pathogenic microbe; programs; response; skin microbiome; skin wound; therapeutic target; tissue repair; transcriptome; transcriptome sequencing; transcriptomics; virtual; wound; wound healing; wound response; wound treatment

PROJECT SUMMARY Improved therapeutic approaches are needed for non-healing wounds, as they present a major challenge to the healthcare system by increasing treatment costs as well as rates of morbidity and mortality. The skin microbiome exists at the interface of all cutaneous wounds, but its potential as a novel therapeutic target remains untapped. Therefore, our long-term goal is to understand host-microbial interactions during wound healing, so we may leverage these mechanisms to identify wound healing treatments. To initiate this investigation, our lab previously performed a metagenomic analysis on wound samples from non-infected diabetic foot ulcers (DFUs) and unearthed a surprisingly prevalent wound inhabitant Alcaligenes faecalis. Very little is known about this species in the context of wounds, so we sought to investigate the effect of A. faecalis on wound healing. We observed the surprising finding that treating wounds with A. faecalis accelerates the rate of wound healing in vivo and activates a pro-epithelialization phenotype in keratinocytes. Thus, the central goal of this study is to identify the mechanism by which A. faecalis mediates accelerated wound healing. Pro- inflammatory cytokine signaling is necessary to promote re-epithelialization, and IL-6 in particular has been shown to induce activation of keratinocytes during healing. Therefore, I tested if A. faecalis induced a cytokine response in keratinocytes and found robust IL-6 production after treatment with A. faecalis conditioned media. A primary mechanism by which bacteria can modulate host responses is through production of secreted molecules. I found that sterile supernatant of A. faecalis, rather than bacterial-cell surface molecules, promotes keratinocyte migration and IL-6 production. Together, these findings lead to my hypothesis that A. faecalis produces secreted molecules that improve re-epithelialization by enhancing keratinocyte IL-6 signaling. Aim 1 will determine the host mechanisms of A. faecalis-induced accelerated healing through a combination of wound healing assays and transcriptional profiling. Aim 2 will identify the microbial genetic determinants of A. faecalis necessary to produce the pro-healing secreted molecule. I will use comparative genomics approach to identify genomic loci that segregate with a pro-wound healing phenotype. To complete this aim, I will leverage our collection of 44 A. faecalis clinical DFU isolates and perform a genomic multiple alignment. I will pair this genetic screen with a phenotypic screen using wound healing assays to determine which genomic locus segregates with the pro-healing phenotype. In conjunction with these experimental aims, I will also engage in a rigorous training plan at the University of Pennsylvania under the guidance of the MD/PhD Program and my PhD advisor. This training plan will foster my development as a future physician scientist through the following goals: strengthen independence as an investigator, develop computational and wet lab techniques, expand knowledge in the microbiome and wound healing fields, improve scientific communication, and engage in mentorship.

项目摘要 非愈合伤口需要改进的治疗方法，因为它们呈现了主要的治疗方法 通过增加治疗成本以及发病率和死亡率来挑战医疗保健系统。 皮肤微生物组存在于所有皮肤伤口的界面上，但其潜力是一种新型的治疗性 目标仍未开发。因此，我们的长期目标是了解宿主 - 微生物相互作用 伤口愈合，因此我们可以利用这些机制来识别伤口愈合治疗。启动这一点 调查，我们的实验室先前对未感染的伤口样品进行了宏基因组分析 糖尿病足溃疡（DFUS），并发掘出令人惊讶的居民alcaligenes粪便。非常 在伤口的背景下，对该物种知之甚少，因此我们试图研究粪曲霉的作用 关于伤口愈合。我们观察到了一个令人惊讶的发现，即用烟曲霉的伤口加速了速率。 体内伤口愈合并激活角质形成细胞中的促临界表型。因此，中心目标 这项研究的目的是确定烟曲霉介导加速伤口愈合的机制。 pro 炎症性细胞因子信号传导对于促进重新上皮化是必要的，尤其是IL-6是 显示在愈合过程中诱导角质形成细胞的激活。因此，我测试了烟曲霉是否诱导细胞因子 角质形成细胞的反应，并在用烟曲霉条件培养基治疗后发现了强大的IL-6产生。 细菌可以调节宿主反应的主要机制是通过生产分泌的 分子。我发现烟曲霉的无菌上清液，而不是细菌表面分子促进 角质形成细胞迁移和IL-6产生。这些发现一起导致了我的假设，即 产生分泌的分子，通过增强角质形成细胞IL-6信号传导来改善重新上皮化。目标1 将通过结合结合 伤口愈合测定法和转录分析。 AIM 2将识别A的微生物遗传决定因素。 产生亲治的分泌分子所必需的粪便。我将使用比较基因组学方法 识别基因组基因座，以促进愈合的愈合表型分离。为了完成这个目标，我将利用 我们收集了44张A.粪便临床DFU分离株并进行基因组多重比对。我会配对 使用伤口愈合测定法确定哪个基因组基因座 与亲治表型分离。 结合这些实验目标，我还将在 宾夕法尼亚大学在医学博士/博士计划的指导下和我的博士顾问。这个培训计划 将通过以下目标来促进我作为未来医师科学家的发展：加强 独立作为研究者，开发计算和湿实验室技术，扩大知识 微生物组和伤口愈合领域，改善科学交流并参与指导。