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.

生发中心是次级淋巴器官中控制B细胞克隆形成的微结构部位 扩增以产生用于实现体液免疫的高亲和力抗体。GC结构受损， 阻塞性睡眠呼吸暂停（OSA）患者，导致感染个体免疫反应的缺陷。 B细胞通过GC反应发育的作用已得到充分证实。B细胞免疫代谢是 这对满足快速扩散的能源需求至关重要。OSA患者表现出与代谢相关的 综合症和易患流感。然而，OSA+和OSA+的B细胞中代谢轨迹的协调是不一致的。 OSA-患者仍然不清楚。迫切需要破译B细胞免疫代谢 在GC中的单细胞水平用于鉴定OSA患者免疫系统中的代谢缺陷 使他们容易受到威胁生命的感染。因此，该项目将利用最近开发的 空间分辨代谢分析框架（3D-SMF），以绘制B细胞亚群及其在 扁桃体组织和类器官。我们的长期目标是产生B细胞的单细胞代谢见解 OSA+和OSA-患者响应流感的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像素的空间分辨B细胞免疫代谢。我们 假设B细胞亚群的代谢轨迹和空间分布在OSA+中是有缺陷的， 与OSA-组织和OSA-扁桃体衍生类器官中的流感反应相比， OSA+扁桃体衍生的类器官。该假设的依据是3D-SMF数据，显示 天然扁桃体组织中GC中脂肪酸的消耗和富集，以及最近的证据表明， 阻塞性睡眠呼吸暂停综合征患者易受流感感染与代谢综合征有关。核心假设是 通过追求两个具体目标进行测试。目的1将提供一个整体的了解脂质相关的 人OSA+和OSA-扁桃体组织中B细胞亚群的免疫代谢（各n=10）和工程化 扁桃体类器官目的2将定义扁桃体中B细胞亚型的代谢轨迹建模如何不同 OSA+和OSA-供体中暴露于流感抗原的类器官。为了实现这些目标，3D-SMF和 多重细胞因子基因表达谱将用于分析B细胞免疫代谢， 假时间B细胞发育建模和慢性乙型肝炎患者B细胞亚群的纵向代谢轨迹比较 生物材料基扁桃体类器官。该项目建立了一个跨学科的团队， 空间组学、生物材料、小儿OSA和生物信息学。建议的应用是创新的 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， 从传统的关注T细胞和B细胞共培养转向B细胞代谢的差异， 生理学相关的和动态的OSA+和OSA-离体扁桃体类器官。这项研究意义重大 因为它定义了B细胞的免疫代谢，以了解为什么OSA患者容易感染流感。