基于单细胞测序研究CXCL14和DCs亚群细胞失衡参与COPD向NSCLC转化的免疫机制OA
Single-cell sequencing reveals CXCL14 upregulation and dendritic cell subset imbalance in the immunologic transition from chronic obstructive pulmonary disease to non-small cell lung cancer
目的 探讨趋化因子配体14(chemokine ligand 14,CXCL14)上调和树突状细胞(dendritic cells,DCs)亚群细胞失衡参与慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)向非小细胞肺癌(non-small cell lung cancer,NSCLC)转化的免疫机制.方法 临床招募无慢性疾病且未服用常规药物的吸烟健康对照者(SmHC组),稳定期COPD患者(stCOPD组),急性加重期COPD患者(exCOPD组)和首次确诊的吸烟NSCLC患者但尚未接受任何治疗(NSCLC组),各3例.采集其外周静脉血样和痰液样本.抗体微阵列芯片检测痰液中的细胞因子水平.qRT-PCR法对微阵列芯片中的细胞因子mRNA表达水平进行测定和验证.单细胞质谱分析对外周血单核细胞(peripheral blood mononuclear cells,PBMCs)中的免疫细胞进行计数、元聚类和均匀流形逼近与投影降维(UMAP)分析.流式细胞术对浆细胞样树突状细胞(plasmacytoid dendritic cells,pDCs)(CD64+DCs)、组成型 DC1s(conventional DC1s,cDC1s)(CD64+MAR-1+CXCL14+DCs)和组成型cDC2s(CD64+CXCL14+DCs)的表面标志物表达进行验证;对pDCs、cDC1s和cDC2s在SmHC组、stCOPD组、exCOPD组和NSCLC组PBMCs的分化DCs中的占比进行计数测定.结果 与stCOPD组相比,NSCLC组患者痰液样本中细胞因子水平显著差异的有:白细胞介素-6(interleukin 6,IL-6)、程序性死亡受体配体-1(programmed death ligand 1,PD-L1)、CD86、Toll 样受体 2(Toll-like receptor 2,TLR-2)、程序性死亡受体-1(programmed death 1,PD-1)、淋巴细胞活化基因-3(lymphocyte activation gene 3,LAG-3)、白细胞介素-1α(interleukin-1 α,IL-1α)、骨形态发生蛋白4(bone morphogenetic protein 4,BMP-4)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)、CXCL14、巨噬细胞集落刺激因子(macrophage colony-stimulating factor receptor,M-CSF)显著上调(P＜0.01);细胞毒性 T 淋巴细胞相关抗原 4(cytotoxic T-lymphocyte-associ-ated antigen 4,CTLA-4)、T 细胞免疫球蛋白黏液素 3(T cell immunoglobulin and mucin-domain-containing-3,TIM-3)上调(P＜0.05);骨形态发生蛋白6(bone morphogenetic protein 6,BMP-6)、干扰素-γ诱导的单核因子(monokine induced by interferon-γ,MIG)显著下调(P＜0.01).与 exCOPD 组相比,NSCLC 组 CXCL14 mR-NA相对表达水平上调(P＜0.05).单细胞质谱分析确定了 12个主要亚群,其中pDCs(3.92％)、cDC1s(3.85％)、cDC2s(2.63％);PD-1和PD-L1在上述3个细胞亚群中呈不同程度上调表达.与stCOPD组相比,exCOPD组和NSCLC组中cDC1s亚群细胞和cDC2s亚群细胞占比(％)升高(P＜0.05).与exCOPD组相比,NSCLC组MAR-1和CXCL14在cDC1s细胞中的相对表达水平升高(P＜0.05),CXCL14在cDC2s细胞中的相对表达水平升高(P＜0.05),CD64在cDC1s细胞、cDC2s细胞和pDCs细胞中的相对表达水平升高(P＜0.05),cDC1s亚群细胞和cDC2s亚群细胞占比(％)升高(P＜0.05).结论 CXCL14上调和DCs亚群细胞占比失衡参与COPD向NSCLC的转化.
Objective To elucidate the immunologic mechanisms by which upregulation of chemokine ligand 14(CXCL14)and imbalance of dendritic cell(DC)subsets contribute to the transition from chronic obstructive pulmonary disease(COPD)to non-small cell lung cancer(NSCLC).Methods Clinical participants were recruited into four groups:smoking healthy controls(SmHC),stable COPD(stCOPD),acute exacerbation COPD(exCOPD),and newly diagnosed,treatment-naïve smoking NSCLC(NSCLC),with three subjects in each group.Peripheral venous blood and induced sputum samples were collected.Cytokine profiles in sputum were assessed using antibody microarray chips.Dif-ferential mRNA expression of cytokines was validated by qRT-PCR.Single-cell mass cytometry was applied to profile peripheral blood mononuclear cells(PBMCs),enabling immune-cell quantification,metaclustering,and UMAP-based dimensionality reduction.Flow cytometry validated surface markers of plasmacytoid dendritic cells(pDCs;CD64+DCs),conventional DC1s(cDC1s;CD64+MAR-1+CXCL14+DCs),and conventional DC2s(cDC2s;CD64+CXCL14+DCs),and quantified subset proportions among differentiating DCs in the four study groups.Results Compared with the stCOPD group,the NSCLC sputum samples demonstrated significant upregulation of interleukin 6(IL-6),programmed death ligand 1(PD-L1),CD86,Toll-like receptor 2(TLR-2),programmed death 1(PD-1),lymphocyte activa-tion gene 3(LAG-3),interleukin-1α(IL-1α),bone morphogenetic protein 4(BMP-4),transforming growth fac-tor-β1(TGF-β1),CXCL14,and macrophage colony-stimulating factor receptor(M-CSF)(P＜0.01),as well as cytotoxic T-lymphocyte-associated antigen 4(CTLA-4)and T cell immunoglobulin and mucin-domain-containing-3(TIM-3)(P＜0.05).Bone morphogenetic protein 6(BMP-6)and monokine induced by interferon-γ(MIG)were significantly downregulated(P＜0.01).CXCL14 mRNA levels were elevated in NSCLC compared with exCOPD(P＜0.05).Single-cell mass cytometry identified 12 major immune-cell metaclusters,including pDCs(3.92％),cDC1s(3.85％),and cDC2s(2.63％).PD-1 and PD-L1 exhibited varying degrees of upregulation across these DC sub-sets.Compared with stCOPD,the proportions of cDC1s and cDC2s increased in exCOPD and NSCLC(P＜0.05).Com-pared with exCOPD,NSCLC patients showed higher expression of MAR-1 and CXCL14 in cDC1s(P＜0.05),increased CXCL14 expression in cDC2s(P＜0.05),and elevated CD64 expression in cDC1s,cDC2s,and pDCs(P＜0.05).Proportions of cDC1s and cDC2s were also significantly increased(P＜0.05).Conclusion CXCL14 upregulation and imbalance of DC subsets,particularly cDC1 and cDC2 expansion accompanied by immunoregulatory marker elevation,may play critical roles in driving the immunologic shift from COPD to NSCLC.
甄丽芳;邵杰;汪雅琴;李瑞丹;秦雅红;吴水淼
兵器工业卫生研究所(兵器工业总医院)呼吸与危重症医学科(陕西西安 710065)兵器工业卫生研究所(兵器工业总医院)呼吸与危重症医学科(陕西西安 710065)兵器工业卫生研究所(兵器工业总医院)呼吸与危重症医学科(陕西西安 710065)兵器工业卫生研究所(兵器工业总医院)呼吸与危重症医学科(陕西西安 710065)兵器工业卫生研究所(兵器工业总医院)呼吸与危重症医学科(陕西西安 710065)渭南市中心医院呼吸与危重症医学科(陕西渭南 714000)
医药卫生
慢性阻塞性肺疾病非小细胞肺癌单细胞测序树突状细胞CXCL14趋化因子
chronic obstructive pulmonary diseasenon-small cell lung cancersingle-cell sequencingden-dritic cellsCXCL14 chemokine
《广东医学》 2026 (1)
45-54,10
陕西省卫生健康委科研基金项目(2021D014)
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