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

调节性b细胞（Bregs）是免疫耐受的重要组成部分，随着年龄和癌症的增长，它们在数量和表型上发生改变1-3。与其他B细胞群不同，Bregs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破坏Bregs的功能。线粒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Bregs中与年龄相关的线粒体/代谢变化如何影响它们向浆细胞的分化及其抑制功能。Aim-1。定义与年龄相关的b细胞和Bregs代谢变化（Mauri-Bradford UCL）。20至70岁以上供者的PBMC/脾细胞将被抗体染色，确定b细胞亚群，并通过多参数流式细胞术分析代谢途径。b细胞糖酵解和呼吸速率也将由海马评估。Aim-2。定义Bregmetabolic转录组中与年龄相关的变化。(Fraternali-Ng国王)。我们将从年轻（20-25岁）和老年（70岁以上）（n=3）成年人的脾脏/血液中分选Bregs，并进行单细胞rna测序。我们提出了一种多学科的方法，通过使用最先进的计算算法分析生成的scrna序列数据集，然后进行实验验证，来询问Breg代谢的相关变化。通过计算方法确定的关键Breg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/药理学方法干扰基因/代谢物驱动因素，以总结或挽救与年龄相关的Breg缺陷（Mauri-Bradford’s）。我们期望对B细胞、Breg代谢及其在衰老过程中的失调有更好的了解。意义：Bregs是免疫系统的重要组成部分，了解衰老过程中其代谢需求的变化如何影响其调节功能对于全面了解老年免疫稳态至关重要。

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.