Regulatory B cell metabolism and ageing: Integrating experiment and data analysis

调节性 B 细胞代谢与衰老：实验与数据分析相结合

• 批准号: 2723178
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2723178
• 关键词: Regulatory; cell; metabolism; ageing; Integrating

Regulatory B-cells (Bregs) are an important component of immune-tolerance, they arenumerically and phenotypically altered with increasing age and in cancer1-3. Bregdifferentiation and function, unlike other B cell populations, requires oxidativephosphorylation and high mitochrondria capacity and we have shown that interferencewith these metabolic pathways abrogates Bregs function4. Declining mitochondrialquality and metabolic activity are associated with normal aging, correlating with thedeclining of the immune function5. Here we will investigate how age-relatedmitochondrial/metabolic changes in Bregs affect their differentiation into plasma cellsand their suppressive function. Aim-1. Define age-related metabolic changes in B-cellsand Bregs (Mauri-Bradford UCL). PBMC/splenocytes from 20 to 70+ year-old donors willbe stained with antibodies defining B-cell subsets and metabolic pathways will beanalysed by multiparameter flow cytometry. B-cell glycolysis and respiration rates willalso be assessed by seahorse. Aim-2. Define age-related changes in the Bregmetabolic transcriptome. (Fraternali-Ng King's). We will sort Bregs from thespleens/blood of young (20-25y/o) and aged (70+) (n=3) adults and perform singlecell-RNA-sequencing. We propose a multi-disciplinary approach to interrogate agerelatedchanges of Breg metabolism through analysing the generated scRNA-seqdatasets using state-of-the-art computational algorithms followed by experimentalvalidation. Descriptions of key Breg cellular/metabolomic states identified bycomputational methods will be substantiated, integrating different biologicalannotations (protein-protein interactions, pathways, metabolomics) (Fraternali-Ng's).We will also perturb gene/metabolite drivers with genetic/pharmacological approachesto recapitulate or rescue age-associated Breg deficits (Mauri-Bradford's). We anticipateimproved understanding of B cells, Breg metabolism and its deregulation in ageing.Significance: Bregs are an essential component of the immune system andunderstanding how changes in their metabolic requirements in ageing affects theirregulatory function is crucial to a full understanding of immune homeostasis in oldage.

调节性B细胞（BCRs）是免疫耐受的重要组成部分，随着年龄的增长和癌症的发生，它们在数量和表型上都发生了变化1 -3。与其他B细胞群不同，Breg的分化和功能需要氧化磷酸化和高线粒体能力，我们已经证明，干扰这些代谢途径会消除Breg的功能4。身体质量和代谢活动的下降与正常衰老有关，与免疫功能的下降有关5。在这里，我们将研究年龄相关的线粒体/代谢变化如何影响他们的分化为浆细胞和他们的抑制功能。目标一定义年龄相关的B细胞代谢变化和B细胞（Mauri-Bradford UCL）。20 - 70+岁供体的PBMC/脾细胞将用定义B细胞亚群的抗体染色，代谢途径将通过多参数流式细胞术分析。海马的B细胞糖酵解和呼吸速率也将被评估。目标二定义Breg代谢转录组中与年龄相关的变化。（Fraternali-Ng King's）.我们将从年轻人（20- 25岁）和老年人（70岁以上）（n=3）的脾/血液中分选出Bcirrhosis，并进行erceell-RNA测序。我们提出了一种多学科的方法来询问年龄相关的变化的布雷格代谢，通过分析所产生的scRNA-seq数据集使用国家的最先进的计算算法，然后通过实验验证。通过计算方法确定的关键布雷格细胞/代谢组学状态的描述将得到证实，整合不同的生物学注释（蛋白质-蛋白质相互作用，途径，代谢组学）（Fraternali-Ng's）。我们还将用遗传/药理学方法扰乱基因/代谢物驱动因素，以重现或挽救与年龄相关的布雷格缺陷（Mauri-Bradford's）。我们期望提高对B细胞、布雷格代谢及其在老龄化中的失调的理解。意义：布雷格是免疫系统的重要组成部分，了解其代谢需求在老龄化中的变化如何影响其调节功能对于充分理解老年免疫稳态至关重要。