自体造血干细胞移植后长生存多发性骨髓瘤患者的骨髓免疫表达谱特征分析

Investigation of the immune profile of multiple myeloma patients achieving long-term survival after autologous stem cell transplantation

目的探讨与多发性骨髓瘤(MM)患者长生存相关的骨髓免疫微环境特征。方法在中山大学附属第一医院明确诊断为新诊断MM并接受新药诱导续贯自体造血干细胞移植以及免疫调节剂维持治疗的随访队列中, 在2019年8月至2020年5月横断面研究期间, 通过高通量NanoString技术比较16例无进展生存≥5年患者[长生存组, 其中微小残留病(MRD)持续阴性亚组9例、MRD持续阳性亚组7例]与5例疾病进展患者(疾病进展组)骨髓中免疫细胞亚群及770个与免疫相关标志物的RNA表达。通过细胞特征性基因表达水平综合计算各免疫细胞功能分值, 从而间接得出各免疫细胞亚群比例。统计学分析主要采用Mann-WhitneyU检验、Kruskal Wallis秩和检验。结果 (1)长生存组患者骨髓中中性粒细胞比例较疾病进展组显著增加[功能分值:13.61(13.33, 14.25)比12.93(12.58, 13.38);Z=2.31, P=0.021];与MRD持续阴性长生存亚组相比, MRD持续阳性长生存亚组患者骨髓肥大细胞显著增多[功能分值:7.09(6.49, 8.57)比6.03(5.18, 6.69);H=2.18, P=0.029]。(2)相比疾病进展组, 长生存组显著上调基因4个(CTSG、IFIT2、S100B、CHIT1), 显著下调基因6个(C4B、TNFRSF17、CD70、IRF4、C2、GAGE1);相比MRD持续阴性长生存亚组, MRD持续阳性长生存亚组显著上调基因CMA1, 下调基因10个(ISG15、OAS3、MX1、IFIT2、DDX58、SIGLEC1、CXCL10、IL1RN、SERPING及TNFSF10), 其中前5个基因均为干扰素反应通路相关基因。结论骨髓中性粒细胞增多、肥大细胞增多可能与移植后MM患者长生存相关。干扰素反应通路激活可能是MRD持续阴性长生存MM患者骨髓免疫微环境的特征之一。

Objective To identify the characteristics of the bone marrow immune microenvironment associated with long-term survival in multiple myeloma(MM)patients.Methods In the follow-up cohort of patients with newly diagnosed MM and who received“novel agent induction therapy and subsequent autologous stem cell transplantation and immunomodulator maintenance therapy”in the First Affiliated Hospital of Sun Yat-sen University,a cross-sectional study was carried out between August 2019 and May 2020.Using NanoString technology,the RNA expression of 770 bone marrow immune-related markers was compared between 16 patients who had progression-free survival≥5 years and 5 patients with progressive disease.Among the 16 patients who achieved long-term survival,9 achieved persistent minimal residual disease(MRD)negative while the other 7 had persistent positive MRD.The functional scores of each kind of immune cells were calculated based on the expression level of characteristic genes,so as to indirectly obtained the proportion of each immune cell subset.The Mann-Whitney U test and the Kruskal Wallis test were used for statistical analysis.Results The proportion of neutrophils was significantly higher in long-surviving MM patients than in patients with progressive disease[functional scores,13.61(13.33,14.25)vs.12.93(12.58,13.38);Z=2.31,P=0.021].Among long-surviving patients,those who were MRD-positive had a significantly greater number of mast cells compared with those who were MRD-negative[functional scores,7.09(6.49,8.57)vs.6.03(5.18,6.69);H=2.18,P=0.029].Compared with patients with progressive disease,four genes(CTSG,IFIT2,S100B,and CHIT1)were significantly downregulated and six(C4B,TNFRSF17,CD70,IRF4,C2,and GAGE1)were upregulated in long-surviving patients.Among long-surviving patients,only gene CMA1 was significantly upgraded,10 genes(ISG15,OAS3,MX1,IFIT2,DDX58,SIGLEC1,CXCL10,IL1RN,SERPING and TNFSF10)were significantly downregulated in the MRD-positive group compared with that in the MRD-negative group,the first 5 of which are related to the interferon response pathway.Conclusions The increased neutrophil and mast cell numbers may be related to long-term survival in MM.Interferon signaling activation may be a key bone marrow immune profiling feature for MRD-negative,long-surviving patients with MM.