Th17 extracellular trap-mediated antimicrobial host defense in acne vulgaris

寻常痤疮中 Th17 细胞外陷阱介导的抗菌宿主防御

• 批准号: 10698121
• 负责人: George W Agak
• 金额: $ 42.47万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-06 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10698121
• 关键词: Acne; Acne Vulgaris; Affect; Affinity; Antigen-Presenting Cells; Bacteria; Binding; Biological; Biopsy; Carbohydrates; Cell Differentiation process; Cells; Chromatin; Chronic; Clone Cells; Complex; Coupled; Cutaneous; DNA; Data; Defense Mechanisms; Dendritic Cells; Development; Disease; Education; Equilibrium; Exposure to; Future; Gene Expression Profile; Genes; Goals; Helper-Inducer T-Lymphocyte; Histone H2B; Histones; Homeostasis; Host Defense; Human; IL17 gene; Image; Immune; Immune response; Immune system; Immunologic Techniques; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Interferons; Interleukin-10; Journals; Lesion; Ligands; Lipoproteins; Mass Spectrum Analysis; Mediating; Medical; Metagenomics; Methodology; Modeling; Molecular; Nonlytic; Nucleotides; Pathway interactions; Patients; Persons; Play; Probiotics; Proteomics; Psychological Impact; Psychological Stress; Public Health; Research; Resolution; Role; Scanning Electron Microscopy; Signal Transduction; Skin; Specificity; Staphylococcus aureus; Stimulus; T-Lymphocyte; TLR1 gene; TLR2 gene; TLR6 gene; Testing; antimicrobial; antimicrobial peptide; cell type; clinical investigation; design; disability; dysbiosis; experimental study; extracellular; genetic element; genetic signature; improved; innovation; insight; keratinocyte; lipidomics; microbial; microbiome; microbiome sequencing; monocyte; neutrophil; novel; novel strategies; novel therapeutic intervention; pathogen; pathogenic bacteria; potential biomarker; receptor; response; self esteem; single-cell RNA sequencing; skin disorder; skin lesion; skin microbiome; targeted treatment; therapeutic target; transcriptomics; whole genome

PROJECT SUMMARY Acne vulgaris, or acne, is a disease of the pilosebaceous unit (PSU). Acne is ranked third among chronic skin diseases for causing disability and medical expense and is a major cause of psychological stress in young people. One factor contributing to acne is Cutibacterium acnes, the major bacterial species in the PSU. Using C. acnes as a model to study the interaction between the microbiome and the host immune response, we: 1) previously demonstrated the presence of IL-17 in acne skin lesions and 2) recently discovered that C. acnes phylotypes associated with acne (CA) or healthy skin (CH) differentially regulate the fate of TH17 cells to develop into non-antimicrobial (n-AMTH17) and antimicrobial (AMTH17) subsets. AMTH17 cells release T cell extracellular traps (TETs) and directly kill C. acnes and other bacteria pathogens. To date, the mechanisms by which C. acnes phylotypes induce TETs and their biological impact in acne are unknown. Therefore, our proposed research is innovative because we will define the immune landscape of the acne lesions, provide mechanistic insights into the biological impact of TH17-TET formation in acne, and identify novel immune pathways and potential biomarkers that can be targeted for acne therapy. This is important as future strategies could be developed to modulate TH17 function. Additionally, maintaining the balance among the different phylotypes of C. acnes may represent a strategy for novel probiotic design. The identification of C. acnes ligands will further elucidate the specificity of host receptors involved in microbial surveillance and lead to the development of novel therapeutic approaches skin diseases caused by dysbiosis. to control acne and other human Our central hypothesis is that innate activation of TH17 cells leads to induction of antimicrobial mechanisms, including TETs, which contribute to host defense against C. acnes and other bacteria. To elucidate this, we will determine the antimicrobial mechanisms of AMTH17 cells against C. acnes (Aim 1), investigate the role of AMTH17 cells in acne inflammation (Aim 2), and identify the C. acnes ligands that induce AMTH17 differentiation (Aim 3). Our preliminary findings support the premise that healthy skin commensals are critical to the education of our immune system and our overall defense against pathogens. Our strategy will include: classical immunological techniques involving T cell cloning to dissect the immune effector functions that underlie TH17-mediated antimicrobial host defense; microbiome sequencing of C. acnes phylotypes that inhabit donor biopsies to define how interactions within the skin microbiome and the host immune response influences acne development; high-resolution time-lapse imaging to define mechanisms of TET release and chromatin dynamics that occur during TET formation; scanning electron microscopy to delimit extracellular trap formation in human T cells; and state-of-the-art single cell RNA-seq experiments to compute T cell gene signatures and to define the T cell types and immune circuits present in acne lesions. Additional state-of-the- art methodologies include the use of high affinity capture of cellular interactomes, coupled with mass spectrometry-based proteomics, lipidomics and carbohydrate HPAEC-PAD analysis to identify C. acnes ligands and endogenous complexes. This contribution is significant as, our studies will make significant conceptual advances in our understanding of T cell–antimicrobial defense mechanisms, and allow major advances in the understanding of the immune networks in acne that can be targeted for therapy

项目摘要 痤疮粉刺或痤疮是焦虑单元（PSU）的一种疾病。痤疮排名第三 慢性皮肤疾病导致残疾和医疗费用，是造成心理压力的主要原因 在年轻人中。造成痤疮的一个因素是痤疮痤疮，这是痤疮，这是 PSU。使用C. acnes作为模型来研究微生物组与宿主免疫之间的相互作用 回应，我们：1）先前证明了痤疮皮肤病变中IL-17的存在，最近2） 发现与痤疮（CA）或健康皮肤（CH）相关的C.痤疮型痤疮型型细胞型会差异调节 Th17细胞的命运形成非抗菌（N-AMTH17）和抗微生物（AMTH17）亚群的命运。 Amth17细胞 释放T细胞外陷阱（TET），直接杀死痤疮梭菌和其他细菌病原体。 迄今 痤疮是未知的。因此，我们提出的研究具有创新性，因为我们将定义免疫力 痤疮病变的景观，提供对Th17-Tet形成生物学影响的机械见解 痤疮，并鉴定可用于痤疮治疗的新型免疫途径和潜在的生物标志物。 这很重要，因为可以制定未来的策略来调节TH17功能。另外，维护 痤疮藻的不同系统型之间的平衡可能代表了新型益生菌设计的策略。 痤疮梭菌配体的鉴定将进一步阐明涉及微生物的宿主受体的特异性 监视并导致新型治疗方法的发展 由营养不良引起的皮肤疾病。 控制痤疮和其他人 我们的中心假设是，Th17细胞的先天激活导致抗菌诱导 包括TET在内的机制，有助于宿主防御痤疮梭菌和其他细菌。到 阐明这一点，我们将确定针对痤疮梭菌的Amth17细胞的抗菌机制（AIM 1）， 研究Amth17细胞在痤疮注射中的作用（AIM 2），并鉴定出影响的痤疮梭菌配体 Amth17差异化（AIM 3）。我们的初步发现支持了健康的皮肤儿童的前提 对我们的免疫系统的教育和对病原体的整体防御至关重要。我们的策略将 包括：涉及T细胞克隆以剖析免疫效应功能的经典免疫技术 这是Th17介导的抗菌宿主防御的基础；痤疮甲状腺梭状芽胞杆菌的微生物组测序 居住在供体活检中，以定义皮肤微生物组和宿主免疫反应中的相互作用 影响痤疮的发展；高分辨率的延时成像，以定义TET释放的机制和 在TET形成期间发生的染色质动力学；扫描电子显微镜以界定细胞外陷阱 在人T细胞中形成；和最先进的单细胞RNA-seq实验以计算T细胞基因 特征并定义痤疮病变中存在的T细胞类型和免疫循环。额外的最新 ART方法包括使用高亲和力捕获细胞相互作用，并加上质量 基于光谱法的蛋白质组学，脂质组学和碳含碳水化合物HPAEC-PAD分析，以鉴定痤疮梭菌 配体和内源复合物。这项贡献很重要，因为我们的研究将使 我们对T细胞 - 抗微生物防御机制的理解的概念进步，并允许主要 了解可以用于治疗的痤疮免疫网络的进步