PO.CL05.09 · 临床研究
单细胞转录组学揭示CD4+ T细胞与B细胞之间的功能性相互作用，驱动携带种系致病性BRCA1变异健康女性的活化外周免疫表型
Single-cell transcriptomics reveal functional interaction between CD4+ T cells and B cells driving the activated peripheral immune phenotype in healthy women living with germline pathogenic BRCA1 variants
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
我们此前基于质谱流式细胞术的结果提示，携带种系致病性BRCA1变异(gpath(BRCA1))的健康女性以及三阴性乳腺癌(TNBC)患者中外周活化CD4+ T细胞和B细胞富集。我们的目的是评估这种活化的转录背景，并研究这两个细胞群之间的相互关系。从5名携带gpath(BRCA1)的健康女性、7名携带gpath(BRCA1)的初治TNBC女性以及7名年龄匹配、无遗传性癌症易感性的健康女性(对照组)中分离外周血单个核细胞。使用Chromium Controller对活的单细胞悬液进行单细胞转录组文库制备。文库在Novaseq 6000仪器上测序。使用CellRanger、Seurat、AUCell、CellChat和Monocle3软件包进行生物信息学分析。质量控制后，分析了100,132个细胞。对CD4+ T细胞、CD8+ T细胞、NK细胞、B细胞和单核细胞进行注释并再聚类为亚群。与对照组相比，gpath(BRCA1)研究组中的B细胞频率降低。促炎性B细胞亚群在健康gpath(BRCA1)携带者中更常见，而活化的Th17极化CD4+ T细胞亚群在健康和携带TNBC的gpath(BRCA1)研究组中均升高。这两个亚群之间的配体-受体相互作用网络分析显示，gpath(BRCA1)携带者中相互作用强度显著增加，并鉴定出介导所观察到的活化外周免疫表型的多种炎性细胞因子。我们的结果证实了携带gpath(BRCA1)健康女性的活化外周免疫特征。这种活化的外周免疫表型以及CD4+ T细胞与B细胞之间的功能性相互作用，可能作为携带gpath(BRCA1)女性精准预防策略的靶点。
查看英文原文 English abstract
Our previous, mass cytometry-based results signalled an enrichment of peripheral activated CD4+ T and B cells in healthy women living with germline pathogenic BRCA1 variants (gpath( BRCA1 )) and in triple-negative breast cancer (TNBC) patients. Our aim was to assess the transcriptional background of this activation and investigate the interrelation between these two cell populations. Peripheral blood mononuclear cells were isolated from five healthy women living with gpath( BRCA1 ), seven women with treatment-naive TNBC living with gpath( BRCA1 ) and seven, age-matched healthy women living without hereditary cancer predisposition (control group). Viable single-cell suspensions were subjected to single-cell transcriptomic library preparation using the Chromium Controller. Libraries were sequenced on a Novaseq 6000 instrument. Bioinformatic analyses were performed using the CellRanger, Seurat, AUCell, CellChat and Monocle3 packages. Following quality control, 100 132 cells were analyzed. CD4+ T cells, CD8+ T cells, NK cells, B cells and monocytes were annotated and subclustered to subpopulations. B cell frequencies in gpath( BRCA1 ) study groups were decreased compared to controls. A pro-inflammatory B cell subpopulation was more frequent in healthy gpath( BRCA1 ) carriers while an activated Th17-polarized CD4+ T cell subpopulation was elevated in both healthy and TNBC-bearing gpath( BRCA1 ) study groups. Ligand-receptor interaction network analysis between these two subpopulations revealed a striking increase in interaction strength in gpath( BRCA1 ) carriers and identified multiple inflammatory cytokines which mediate the observed activated peripheral immune phenotype. Our results confirm the activated peripheral immune profile of healthy women living with gpath( BRCA1 ). This activated peripheral immune phenotype and the functional interactions between CD4+ T cells and B cells might serve as targets during precision prevention approaches in women living with gpath( BRCA1 ).
利益披露 Disclosure
V. Grolmusz, None..
K. Horti-Oravecz, None..
I. Kelemen, None..
I. Likó, None..
A. Bozsik, None..
T. Pócza, None..
J. Papp, None..
S. Pósa, None..
L. S. Pongor, None..
H. Butz, None..
A. Patócs, None.