Investigating the role of Staphylococcus aureus strain-level diversity on neutrophil recruitment and wound healing in a cutaneous leishmaniasis infection

研究皮肤利什曼病感染中金黄色葡萄球菌菌株水平多样性对中性粒细胞募集和伤口愈合的作用

• 批准号: 10313240
• 负责人: Victoria Lovins
• 金额: $ 4.6万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10313240
• 关键词: Amputation; Antibodies; Automobile Driving; Bacteria; Biological Markers; Biopsy; Cells; Chronic; Classification; Clinical; Cutaneous; Cutaneous Leishmaniasis; Data; Disease; Down-Regulation; Epithelial; Epithelial Cells; Failure; Gene Activation; Gene Expression; Genes; Genome; Human; Impaired healing; Impaired wound healing; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Interleukin-8; Leishmania; Leishmania major; Leishmaniasis; Lesion; Measures; Mediating; Messenger RNA; Metadata; Microbe; Migration Assay; Modeling; Mus; Neutrophil Infiltration; Osteomyelitis; Outcome; Parasite Control; Parasites; Parasitic infection; Pathologic; Pathology; Patient-Focused Outcomes; Patients; Phenotype; Phylogenetic Analysis; Process; Proteins; Prunella vulgaris; Quality of life; Reporter; Research; Resolution; Role; Sepsis; Severities; Site; Skin; Skin Tissue; Skin injury; Skin wound healing; Soft Tissue Infections; Staphylococcus aureus; Staphylococcus aureus infection; Testing; Time; Tissues; Tumor-infiltrating immune cells; Virulence; Work; Wound Infection; Wound models; chronic wound; comparative genomics; cytokine; diabetic ulcer; differential expression; genome sequencing; healing; immunopathology; in vivo; insight; keratinocyte; microbial; microbiota; migration; mortality; mouse model; neutrophil; new therapeutic target; novel; pathogen; response; skin lesion; skin microbiome; skin microbiota; transcriptome sequencing; whole genome; wound; wound healing

PROJECT SUMMARY Cutaneous leishmaniasis is a parasitic infection that causes a variable spectrum of disease ranging from single, self-healing lesions to disfiguring chronic lesions that do not heal despite treatment. While the factors driving lesion chronicity are not clear, it is evident that many of the most severe forms of the disease are caused by uncontrolled inflammation rather than high parasite burden. In mice, chronic leishmaniasis lesions are characterized by increased neutrophil accumulation, persistent activation of inflammatory cytokines, and increased infiltration of immune cells to the lesion site, all of which exacerbate tissue damage and delay healing. Our lab has previously shown that colonizing microbes on the skin contribute to immunopathology in murine models, however the role of the skin microbiota in regulating lesion healing is not known. We have found that the most common bacteria colonizing human cutaneous leishmania lesions is Staphylococcus aureus (S. aureus) which delays wound healing in a strain-specific manner across many different pathologies and is a potent neutrophil recruiter. We have also been able to show that S. aureus clinical isolates cultured from human cutaneous leishmaniasis lesions induce secretion of different amounts of neutrophil chemotactic factors such as interleukin-8 (IL-8) by epithelial cells in a wound setting. Therefore, the central hypothesis of my proposal is that S. aureus delays cutaneous leishmaniasis lesion healing through strain-variable neutrophil recruitment and accumulation. In Aim 1, I will determine the strain-specific effect of S. aureus-mediated neutrophil recruitment on wound healing and immunopathology in both a wounding model and in a cutaneous leishmaniasis model using LysM-EGFP mice. In Aim 2, I will identify differences S. aureus neutrophil recruitment and persistence genes across clinical isolates using whole genome sequencing and comparative genomics and associate this data with clinical outcomes and RNA sequencing gene expression from human cutaneous leishmaniasis lesions. These findings will collectively drive understanding of the strain-specific effects of S. aureus infection and neutrophil recruitment on wound healing, uncover biomarkers that can be used to predict patient outcome, and identify novel therapeutic targets in the treatment of cutaneous leishmaniasis.

项目摘要 皮肤利什曼病是一种寄生虫感染，导致疾病范围从 单身的自我修复病变，可毁容尽管治疗，但仍无法治愈的慢性病变。而这些因素 驾驶病变的慢性不清楚，很明显，疾病的许多最严重的形式是引起的 通过不受控制的炎症而不是高寄生虫负担。在老鼠中，慢性利什曼病病变是 其特征是中性粒细胞的积累增加，炎性细胞因子的持续激活和 免疫细胞对病变部位的浸润增加，所有这些病变都会加剧组织损伤并延迟愈合。 我们的实验室以前已经表明，在皮肤上定居微生物有助于鼠的免疫病理学 但是，模型尚不清楚皮肤微生物群在调节病变愈合中的作用。我们发现 最常见的细菌定植人皮肤利什曼原虫病变是金黄色葡萄球菌（S. 金黄色））以特定于应变的方式延迟伤口愈合，在许多不同的病理上，是一种有效的 中性粒细胞招募者。我们还能够证明金黄色葡萄球菌临床分离物从人类 皮肤利什曼病病变诱导不同量的中性粒细胞趋化因子（例如）分泌 白介素8（IL-8）在伤口环境中由上皮细胞。因此，我的提议的核心假设是 金黄色葡萄球菌通过可变异的中性粒细胞募集和 积累。在AIM 1中，我将确定金黄色葡萄球菌介导的中性粒细胞募集的应变特异性作用 在伤口模型和皮肤利什曼病模型中的伤口愈合和免疫病理学上 使用Lysm-EGFP小鼠。在AIM 2中，我将确定差异S.金黄色金黄色的中性粒细胞招募和持久性 使用整个基因组测序和比较基因组学跨临床分离株的基因，并将其关联 来自人皮肤利什曼病病变的临床结果和RNA测序表达的数据。 这些发现将集体推动对金黄色葡萄球菌感染的应变特异性作用的理解和 嗜中性粒细胞在伤口愈合方面招募，发现可用于预测患者预后的生物标志物，以及 确定新的治疗靶标在治疗皮肤利什曼病的治疗中。