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 (GC) 是次级淋巴器官中控制 B 细胞克隆的微观结构位点 产生高亲和力抗体以实现体液免疫的扩增受到损害。 阻塞性睡眠呼吸暂停（OSA）患者，导致感染者免疫反应缺陷。 B 细胞通过 GC 反应发育的作用已得到充分证实。 满足 OSA 患者快速增殖的能量需求至关重要。 然而，OSA+ 和 B 细胞代谢轨迹的协调。 OSA 患者的情况仍不清楚，迫切需要破译 B 细胞免疫代谢。 在 GC 的单细胞水平上识别 OSA 患者免疫系统的代谢缺陷 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技术。 空间分辨代谢分析框架 (3D-SMF)，用于绘制 B 细胞亚群及其代谢图谱 我们的长期目标是获得 B 细胞的单细胞代谢见解。 OSA+ 和 OSA- 患者的 GC 中针对流感的发展 该项目的目标是定义。 我们在固定人体组织和活体组织中逐像素空间分辨 B 细胞免疫代谢。 OSA+ 中 B 细胞亚群的代谢轨迹和空间分布将出现缺陷 与 OSA 组织相比，OSA 扁桃体衍生的类器官对流感的反应能力更强 该假设的基本原理基于 3D-SMF 数据显示。 位于天然扁桃体组织中的GC中脂肪酸的消耗和富集以及最近的证据 OSA 患者易受与代谢综合征相关的流感感染的影响。 目标 1 将提供对脂质相关的完整理解 人 OSA+ 和 OSA- 扁桃体组织（各 n=10）B 细胞亚群的免疫代谢和工程化 目标 2 将定义扁桃体中 B 细胞亚型的代谢轨迹模型有何不同。 为了实现这些目标，3D-SMF 和 OSA+ 供体中暴露于流感抗原的类器官。 多重细胞因子基因表达谱将用于通过以下方法分析 B 细胞免疫代谢： 拟时 B 细胞发育模型和 B 细胞亚群的纵向代谢轨迹比较 该项目建立了一个由来自不同领域的专家组成的跨学科团队。 空间组学、生物材料、儿科 OSA 和生物信息学。 因为它使用尖端技术来定义扁桃体类器官的空间代谢组学和蛋白质组学， 从传统的对 T 细胞和 B 细胞共培养的关注，转向 B 细胞代谢的差异 生理相关且动态的 OSA+ 和 OSA- 扁桃体类器官离体研究具有重要意义。 因为它定义了 B 细胞免疫代谢，以了解为什么 OSA 患者容易感染流感。