Decoding the microbial burden in diabetic foot ulcers

解读糖尿病足溃疡的微生物负荷

• 批准号: 10159971
• 负责人: Elizabeth Anne Grice
• 金额: $ 51.62万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10159971
• 关键词: Adhesions; Alcaligenes; Amputation; Attenuated; Bioinformatics; Biological Markers; Chronic; Clinical; Coculture Techniques; Complication; Complications of Diabetes Mellitus; Corpus striatum structure; Corynebacterium; Coupling; Development; Diabetes Mellitus; Diabetic Foot Ulcer; Enterococcus faecalis; Environmental Pollution; Family suidae; Fractionation; Gene Expression; Genetic; Genomic approach; Genomics; Genotype; Grant; Growth; Immune response; Impaired healing; Impaired wound healing; In Vitro; Infection; Intervention; Knowledge; Lead; Length of Stay; Lower Extremity; Machine Learning; Malignant neoplasm of prostate; Mass Spectrum Analysis; Mediating; Metadata; Microbe; Microbial Biofilms; Modeling; Mus; Neuropathy; Osteomyelitis; Outcome; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Persons; Phenotype; Production; Prospective cohort study; Research; Risk; Role; Series; Shotguns; Skin; Staphylococcus aureus; Staphylococcus aureus infection; Techniques; Testing; Therapeutic; Toxic effect; United States; Variant; Virulence; Wound Infection; Wound models; antimicrobial; base; chronic wound; clinically relevant; cohort; comparative; cost; cytokine; diabetic ulcer; diabetic wound healing; foot; genome analysis; genome sequencing; genome wide association study; healing; improved; improved outcome; in vivo; keratinocyte; malignant breast neoplasm; metagenomic sequencing; microbial; microbial colonization; microbial genomics; microbiota; microorganism interaction; migration; mortality; mouse model; non-healing wounds; novel; pathogen; phenotypic data; porcine model; predict clinical outcome; predictive marker; response; tissue repair; tissue-repair responses; transcriptome sequencing; whole genome; wound; wound closure; wound healing

Chronic, non-healing wounds are common and costly complications of diabetes. Microbial colonization and biofilm formation are hypothesized to impair wound healing and contribute to severe complications such as osteomyelitis and amputation. Although all chronic wounds are colonized with microbiota, its importance, in the absence of clinical infection, is currently unknown. In this competitive renewal, we hypothesize that host response, wound healing, and clinical DFU outcomes are determined by 1) genomic diversification of the wound pathogen Staphylococcus aureus; 2) commensal interactions with wound pathogens; and 3) commensal interactions with the host. In the previous cycle, we developed a shotgun metagenomic sequencing pipeline to analyze the microbiota colonizing neuropathic, non-infected DFU (n=100; the “DFU100” cohort) in a longitudinal prospective cohort study. We observed that strain-level variation of the wound pathogen Staphylococcus aureus was associated with DFU outcomes. Therefore, in Aim 1, we will use a microbial genomic approach and matched clinical isolates from the DFU100 cohort to identify S. aureus genomic determinants of pathogenesis in DFU and their association with clinical outcomes. We also observed that species clinically regarded as “bystanders” (e.g. skin commensals, environmental contaminants) influence the virulence of wound pathogens and tune host tissue repair responses to promote healing in vivo. Aim 2 will determine if a skin commensal, Corynebacterium striatum, tunes the virulence of S. aureus and improves wound healing in murine and porcine models of S. aureus wound infection. Aim 3 is based on our observation that Alcaligenes faecalis wound isolates promote keratinocyte migration, cytokine secretion, and accelerated wound closure in a murine model of diabetic wound healing. We will establish the mechanism and therapeutic potential of A. faecalis-mediated host responses that lead to accelerated wound healing. The proposed research will use cutting-edge, cross-disciplinary approaches to investigate interactions between wound pathogens, wound “bystanders”, and the host; understanding these mechanisms will lead to improved DFU outcomes as our long-term objectives are to 1) develop novel microbiota-based interventions to improve healing that exploit microbial interactions with each other and the host; and 2) identify microbial biomarkers to classify patients at risk of complication.

慢性、无法愈合的伤口是糖尿病常见且代价高昂的并发症。微生物定植和 生物膜的形成被认为会损害伤口的愈合，并会导致严重的并发症，如 骨髓炎和截肢。尽管所有慢性伤口都被微生物区系定植，但它在 没有临床感染，目前尚不清楚。在这场竞争性更新中，我们假设东道主 反应、伤口愈合和临床DFU结果取决于1)基因组的多样性 伤口病原体金黄色葡萄球菌；2)与伤口病原体共生作用；3) 与宿主的共生互动。在前一个周期中，我们开发了一个鸟枪式元基因组学 用于分析定植于神经病理性、非感染性DFU(n=100；DFU100)的微生物区系的测序流水线 队列)进行纵向前瞻性队列研究。我们观察到伤口的应变水平变化 病原体金黄色葡萄球菌与DFU的预后相关。因此，在目标1中，我们将使用 微生物基因组学方法和DFU100队列中匹配的临床分离株鉴定金黄色葡萄球菌 DFU发病机制的基因组决定因素及其与临床结果的关系。我们还观察到 临床上被视为“旁观者”的物种(如皮肤共生、环境污染物)的影响 降低伤口病原体的毒力，调整宿主组织修复反应，促进体内愈合。目标2将 确定一种皮肤共生菌纹状棒杆菌是否调节金黄色葡萄球菌的毒力并改善 金黄色葡萄球菌伤口感染的小鼠和猪模型的伤口愈合。目标3是基于我们的观察 粪产碱菌创伤分离株促进角质形成细胞迁移、细胞因子分泌并加速 糖尿病伤口愈合的小鼠模型中的伤口闭合。我们将建立机制并进行治疗 粪肠弯曲菌介导的宿主反应加速伤口愈合的潜力。建议数 研究将使用前沿的、跨学科的方法来研究伤口之间的相互作用 病原体、伤口“旁观者”和宿主；了解这些机制将导致改善DFU 结果作为我们的长期目标是1)开发新的基于微生物区系的干预措施，以改善 利用彼此和宿主之间的微生物相互作用进行治疗；以及2)识别微生物生物标记物以 对有并发症风险的患者进行分类。