Decoding Spatially Resolved Single Cell Metabolic Trajectory of Tonsil Tissues and Organoids

解码扁桃体组织和类器官的空间分辨单细胞代谢轨迹

• 批准号: 10751125
• 负责人: Ahmet F. Coskun
• 金额: $ 19.68万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-10 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10751125
• 关键词: 3-Dimensional; Antibody Affinity; Antigens; B-Cell Development; B-Lymphocyte Subsets; B-Lymphocytes; Biocompatible Materials; Bioinformatics; Cells; Cellular Metabolic Process; Childhood; Clonal Expansion; Coculture Techniques; Cytokine Gene; Data; Defect; Engineering; Exhibits; Exposure to; Fatty Acids; Gene Expression Profiling; Goals; Human; Humoral Immunities; Immune response; Immune system; Impairment; Individual; Infection; Influenza; Life; Lipids; Maps; Metabolic; Metabolic syndrome; Metabolism; Modeling; Obstructive Sleep Apnea; Organoids; Patients; Physiological; Proliferating; Proteomics; Reaction; Research; Role; Site; Spatial Distribution; Structure; Structure of germinal center of lymph node; T-Lymphocyte; Technology; Testing; Tissues; Tonsil; flu; human tissue; influenza infection; innovation; insight; metabolomics; response; secondary lymphoid organ

Germinal centers (GCs) are the microstructural sites in secondary lymphoid organs that control B cell clonal expansion to produce high-affinity antibodies for achieving humoral immunity. GC structures are impaired in obstructive sleep apnea (OSA) patients, leading the deficiencies in immune responses in infected individuals. The role of B cell development through GC reactions has been well established. B-cell immunometabolism is crucial to meet the energy needs of rapid proliferation. OSA patients exhibited associations with metabolic syndrome and vulnerability to flu. However, the coordination of metabolic trajectories in B cells of OSA+ and OSA- patients is still not clearly understood. There is a critical need to decipher the B cell immunometabolism at the single cell level in GCs for identifying the metabolic defects in the immune system of OSA patients making them prone to life-threatening infections. Thus, this project will leverage the recently developed spatially resolved metabolic profiling framework (3D-SMF) to map B cell subsets and their metabolism in the tonsil tissues and organoids. Our long-term goal is to generate single cell metabolic insights of B cell development in GCs of OSA+ and OSA- patients in response to influenza. The goal of this project is to define spatially resolved B cell immunometabolism pixel-by-pixel in fixed human tissues and living tissues. We hypothesize that metabolic trajectories and spatial distributions of B-cell subsets will be defective in OSA+ compared to OSA- tissues and influenza response in OSA- tonsil-derived organoids will be more competent than OSA+ tonsil-derived organoids. The rationale for this hypothesis is based on the 3D-SMF data showing the depletion and enrichment of fatty acids in GCs located in native tonsil tissues and the recent evidence on OSA patients’ vulnerability to flu infections associated with metabolic syndrome. The central hypothesis will be tested by pursuing two specific Aims. Aim 1 will provide an integral understanding of the lipid-associated immunometabolism in B-cell subsets in human OSA+ and OSA- tonsil tissues (n=10 each) and engineered tonsil organoids. Aim 2 will define how the metabolic trajectory modeling of B cell subtypes differs in tonsil organoids exposed to influenza antigens in OSA+ and OSA- donors. To accomplish these Aims, 3D-SMF and multiplexed cytokine gene expression profiling will be used to analyze B cell immunometabolism through a pseudotime B cell development modeling and longitudinal metabolic trajectory comparisons of B cell subsets in biomaterial-based tonsil organoids. This project builds an interdisciplinary team integrating experts from spatial omics, biomaterials, pediatric OSA, and bioinformatics. The proposed application is innovative because it uses cutting-edge technology to define spatial metabolomics and proteomics of tonsil organoids and shifts from the traditional focus on T cell and B cell co-cultures, toward differences of B cell metabolism in physiologically relevant and dynamic OSA+ and OSA- tonsil organoids ex vivo. This research is significant because it defines B cell immunometabolism to understand why OSA patients are prone to flu infections.

生发中心(GCs)是次级淋巴器官中控制B细胞克隆的微结构部位 扩增产生高亲和力抗体以实现体液免疫。GC结构受损 阻塞性睡眠呼吸暂停(OSA)患者，导致感染者免疫反应缺陷。 通过GC反应发展B细胞的作用已经得到了很好的证实。B细胞的免疫代谢是 对满足快速扩散的能源需求至关重要。阻塞性睡眠呼吸暂停患者与代谢相关 综合症和对流感的脆弱性。然而，OSA+和OSA+在B细胞中代谢轨迹的协调 阻塞性睡眠呼吸暂停综合征患者仍不清楚。迫切需要破译B细胞的免疫代谢 单细胞水平检测阻塞性睡眠呼吸暂停综合征患者免疫系统代谢缺陷 使他们容易受到危及生命的感染。因此，该项目将利用最近开发的 空间分辨代谢谱框架(3D-SMF)定位B细胞亚群及其代谢 扁桃体组织和器官。我们的长期目标是产生对B细胞的单细胞代谢洞察力 OSA+和OSA-患者对流感反应的GC的发展。这个项目的目标是定义 在固定的人体组织和活组织中逐个像素地空间分辨B细胞免疫代谢。我们 假设OSA+患者B细胞亚群的代谢轨迹和空间分布存在缺陷 与OSA组织和流感反应相比，OSA-扁桃体衍生的有机物将更有能力 而不是OSA+扁桃体衍生的有机化合物。这一假设的理论基础是基于3D-SMF数据显示 天然扁桃体组织中GC中脂肪酸的耗竭和富集性及其最新证据 阻塞性睡眠呼吸暂停综合征患者对与代谢综合征相关的流感感染的脆弱性。中心假设将是 通过追求两个具体目标来检验。目标1将提供对与脂质相关的 人OSA+和OSA-扁桃体组织中B细胞亚群的免疫代谢 扁桃体器官。目标2将定义扁桃体中B细胞亚型的代谢轨迹模型如何不同 OSA+和OSA-献血者暴露于流感抗原的有机物。为了实现这些目标，3D-SMF和 多路细胞因子基因表达谱将用于分析B细胞的免疫代谢 中国人B细胞亚群的伪时间发育模型及纵向代谢轨迹比较 以生物材料为基础的扁桃体有机化合物。该项目建立了一个跨学科的团队，整合了来自 空间组学、生物材料、儿科OSA和生物信息学。拟议的应用程序是创新的 因为它使用尖端技术来定义扁桃体器官的空间代谢组学和蛋白质组学 从传统的T细胞和B细胞共培养转向B细胞代谢的差异 体外生理相关和动态的OSA+和OSA-扁桃体有机化合物。这项研究具有重要的意义 因为它定义了B细胞免疫新陈代谢，以理解为什么OSA患者容易感染流感。