Regulation of Skin Immunity by a Tick Bite

蜱虫叮咬对皮肤免疫的调节

• 批准号: 10337568
• 负责人: Joao Pedra
• 金额: $ 23.18万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-01 至 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10337568
• 关键词: Affect; Anesthetics; Animal Model; Animals; Anti-Inflammatory Agents; Anticoagulants; Applications Grants; Arboviruses; Architecture; Area; Arthropods; Bacteria; Bar Codes; Biogenesis; Biology; Biopsy; Bite; Black-legged Tick; Blood; Castor Bean Tick; Cattle; Cavia; Cell Separation; Cells; Communities; Coupled; Dermal; Dermis; Disease; Environment; Epidermis; Etiology; Europe; Flow Cytometry; Future; Gene Expression Profiling; Genes; Hemostatic Agents; Histology; Homeostasis; Human; Immune; Immune response; Immunity; Immunobiology; Immunology procedure; Inflammation; Intervention; Knowledge; Laboratories; Lead; Literature; Mammals; Maps; Measures; Mediating; Molecular; Molecular Analysis; Mus; N-ethylmaleimide-sensitive protein; Organoids; Parasites; Population; Property; Proteins; RNA; RNA Interference; Reagent; Receptor Gene; Regulation; Research; Research Personnel; Resistance; Resources; Role; Saliva; Salivary; Salivary Glands; Salivary Proteins; Seminal; Site; Skin; System; T-Lymphocyte; Technology; Therapeutic; Thinking; Tick-Borne Diseases; Ticks; Travel; United States; Virus; Work; acquired immunity; arthropod-borne; base; extracellular vesicles; fascinate; feeding; follow-up; foot; functional plasticity; immunoregulation; insight; interspecies communication; keratinocyte; microbial; nanovesicle; neglect; new technology; pathogen; receptor; single-cell RNA sequencing; tick bite; tick feeding; tick saliva; tick transmission; tool; transcriptomics; transmission process; vector; vesicular release; wound healing; γδ T cells

Summary/Abstract: Ticks transmit bacteria, viruses and parasites that cause disease in humans and other animals. This phenomenon is partly due to the secretion of redundant and pluripotent salivary proteins that disrupt host homeostasis and alter inflammation upon blood-feeding. Recently, extracellular vesicles, a heterogenous population of nanovesicles that mediate interspecies communication, were shown to facilitate pathogen transmission to mammals. We developed a tick salivary organoid system that mimics extracellular vesicle release. We also manipulated the biogenesis of tick extracellular vesicles by silencing the expression of N-ethylmaleimide-sensitive factor attachment receptor (SNARE) genes through RNA interference. Finally, we provided causality to our findings by showing that tick extracellular vesicles affect resident dendritic epidermal T cells (DETCs) in the skin. How immune cells are targeted by tick nanovesicles in the skin remains unsettled. Whether tick extracellular vesicles have functional plasticity during interspecies relationships remains elusive. In this R21 application, we will explore the central hypothesis that tick extracellular vesicles provide an advantageous skin immune environment for Ixodes scapularis feeding via the DETC-keratinocyte axis. DETCs interact with keratinocytes, which comprise approximately 95% of the skin epidermal layer. In Aim #1 of this proposal, we will determine whether the effect of I. scapularis extracellular vesicles on DETCs is direct or indirect. We will use single cell RNA sequencing coupled with animal models devoid of DETCs to evaluate the directionality of the skin immune response during a tick bite. In Aim #2 of this grant proposal, we will ascertain the architecture of the skin when the biogenesis of tick extracellular vesicles is disrupted. We will measure the immune response of DETCs through skin biopsies via flow cytometry coupled with cell sorting, microbial stimulation and spatial transcriptomics. Spatial transcriptomics combines traditional histology information with single cell gene expression analysis and positional barcoding. Thus, we will construct an architectural map of genes associated with DETCs and keratinocytes during a tick bite. Collectively, this R21 project will underscore the importance of ticks as arthropods of

摘要/摘要：蜱传播细菌、病毒和寄生虫，导致人类和其他动物患病。 动物。这种现象部分是由于多余的多能唾液蛋白的分泌造成的 破坏宿主体内平衡并改变吸血时的炎症。最近，细胞外囊泡 介导种间通讯的异质纳米囊泡群被证明可以促进 病原体传播给哺乳动物。我们开发了一种模仿细胞外的蜱唾液类器官系统 囊泡释放。我们还通过沉默表达来操纵蜱细胞外囊泡的生物发生 通过 RNA 干扰 N-乙基马来酰亚胺敏感因子附着受体 (SNARE) 基因。最后， 我们通过表明蜱细胞外囊泡影响常驻树突来为我们的发现提供因果关系 皮肤中的表皮 T 细胞 (DETC)。皮肤中的蜱纳米囊泡如何靶向免疫细胞仍然存在 不安定。蜱细胞外囊泡在种间关系中是否具有功能可塑性 仍然难以捉摸。在此 R21 应用中，我们将探讨细胞外囊泡的中心假设 通过 DETC 角质形成细胞为肩胛硬蜱提供有利的皮肤免疫环境 轴。 DETC 与角质形成细胞相互作用，角质形成细胞约占皮肤表皮层的 95%。瞄准 该提案的#1，我们将确定 I. scapularis 细胞外囊泡对 DETC 的影响是否是 直接或间接。我们将使用单细胞 RNA 测序结合缺乏 DETC 的动物模型来 评估蜱虫叮咬期间皮肤免疫反应的方向性。在本拨款提案的目标 2 中，我们 当蜱细胞外囊泡的生物发生被破坏时，将确定皮肤的结构。我们将 通过流式细胞术与细胞分选相结合的皮肤活检来测量 DETC 的免疫反应， 微生物刺激和空间转录组学。空间转录组学结合传统组织学 单细胞基因表达分析和位置条形码信息。因此，我们将构造一个 蜱虫叮咬期间与 DETC 和角质形成细胞相关的基因结构图。总的来说，这款 R21 项目将强调蜱虫作为节肢动物的重要性