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Enrichment of circulating myeloma cells by immunomagnetic beads combined with flow cytometry for monitoring minimal residual disease and relapse in patients with multiple myeloma.
Annals of Hematology ( IF 3.5 ) Pub Date : 2019-11-20 , DOI: 10.1007/s00277-019-03833-5 Ningning Wang 1, 2 , Nahom Tesfaluul 3 , Jia Li 1 , Xiaojuan Gao 1 , Shuai Liu 1, 4, 5 , Baohong Yue 1, 4, 5, 6
Annals of Hematology ( IF 3.5 ) Pub Date : 2019-11-20 , DOI: 10.1007/s00277-019-03833-5 Ningning Wang 1, 2 , Nahom Tesfaluul 3 , Jia Li 1 , Xiaojuan Gao 1 , Shuai Liu 1, 4, 5 , Baohong Yue 1, 4, 5, 6
Affiliation
Difficulty in regularly analyzing marrow myeloma cells (MMCs) and low frequency of circulating myeloma cells (CMCs) in blood presents challenges for monitoring minimal residual disease (MRD) in multiple myeloma (MM). We have developed a set of method for enrichment of CMCs by immunomagetic beads (IMB) combined with flow cytometry (IMB-FCM) based on CD38-APC/CD138-APC antibodies in U266-spiked samples and in 122 patient samples. U266 cell capture efficiency of CD38/CD138-IMB-FCM (6.960, 2.574) was 6- and 2-fold higher than that of FCM (1.032), and the sensitivity of FCM and IMB-FCM was 0.01% and 0.001%, respectively. In MM cohort, the positive rate of CMCs by IMB-FCM increased from 60.5~70.0 to 85~87.2% in newly diagnosed/relapsed and partial remission (PR) patients compared with by FCM (P < 0.05). Two complete remission (CR) patients contain certain amounts of CMCs by IMB-FCM while no CMCs and MMCs were detectable by FCM. Patients exhibiting PR and CR upon therapy had much lower CMC and MMC counts than newly diagnosed/relapsed patients (P < 0.005). Based on MRD measurement in BM and PB samples, all FCM-negative BM samples were also paired with FCM/IMB-FCM-negative PB samples among newly diagnosed, relapsed, and PR patients, and FCM-positive BM samples were accompanied by IMB-FCM-positive results in 88% of corresponding PB samples. CMCs strongly associated with other clinical biomarkers of disease burden, including elevated MMCs, β2-MG, sCrea, and DS and ISS stages, and more serious anemia, bone destruction, and renal impairment (P < 0.05). Logistic regression analysis revealed that elevated β2-MG and moderate-to-more anemia were significant risk factors for the presence of CMCs (P < 0.05). As a noninvasive "liquid biopsy" of monitoring MRD, the potential of IMB-FCM for CMC detection may complement or minimize bone marrow aspiration in future treatment of MM patients.
中文翻译:
通过免疫磁珠结合流式细胞仪富集循环中的骨髓瘤细胞,以监测多发性骨髓瘤患者的最小残留疾病和复发。
难以定期分析血液中的骨髓骨髓瘤细胞(MMC)和血液中循环性骨髓瘤细胞(CMCs)的频率低,对监测多发性骨髓瘤(MM)的最小残留病(MRD)提出了挑战。我们已经开发了一套基于U38加标样品和122个患者样品中CD38-APC / CD138-APC抗体的免疫磁珠(IMB)结合流式细胞仪(IMB-FCM)富集CMC的方法。CD38 / CD138-IMB-FCM(6.960,2.574)的U266细胞捕获效率是FCM(1.032)的6倍和2倍,FCM和IMB-FCM的敏感性分别为0.01%和0.001% 。在MM队列中,与FCM相比,新诊断/复发和部分缓解(PR)患者中IMB-FCM对CMCs的阳性率从60.5〜70.0增加到85〜87.2%。两名完全缓解(CR)患者通过IMB-FCM包含一定量的CMC,而FCM未检测到CMC和MMC。与新诊断/复发的患者相比,治疗后出现PR和CR的患者的CMC和MMC计数要低得多(P <0.005)。根据BM和PB样本中的MRD测量,在新诊断,复发和PR患者中,所有FCM阴性BM样本也与FCM / IMB-FCM阴性PB样本配对,并且FCM阳性BM样本伴有IMB- FCM阳性的结果是88%的相应PB样品。CMC与疾病负担的其他临床生物标志物密切相关,包括MMC升高,β2-MG,sCrea以及DS和ISS分期,以及更严重的贫血,骨破坏和肾功能不全(P <0.05)。Logistic回归分析显示,β2-MG升高和中度至重度贫血是存在CMC的重要危险因素(P <0.05)。作为监测MRD的非侵入性“液体活检”,IMB-FCM在CMC检测中的潜力可能会补充或最小化MM患者未来的骨髓抽吸术。
更新日期:2019-11-20
中文翻译:
通过免疫磁珠结合流式细胞仪富集循环中的骨髓瘤细胞,以监测多发性骨髓瘤患者的最小残留疾病和复发。
难以定期分析血液中的骨髓骨髓瘤细胞(MMC)和血液中循环性骨髓瘤细胞(CMCs)的频率低,对监测多发性骨髓瘤(MM)的最小残留病(MRD)提出了挑战。我们已经开发了一套基于U38加标样品和122个患者样品中CD38-APC / CD138-APC抗体的免疫磁珠(IMB)结合流式细胞仪(IMB-FCM)富集CMC的方法。CD38 / CD138-IMB-FCM(6.960,2.574)的U266细胞捕获效率是FCM(1.032)的6倍和2倍,FCM和IMB-FCM的敏感性分别为0.01%和0.001% 。在MM队列中,与FCM相比,新诊断/复发和部分缓解(PR)患者中IMB-FCM对CMCs的阳性率从60.5〜70.0增加到85〜87.2%。两名完全缓解(CR)患者通过IMB-FCM包含一定量的CMC,而FCM未检测到CMC和MMC。与新诊断/复发的患者相比,治疗后出现PR和CR的患者的CMC和MMC计数要低得多(P <0.005)。根据BM和PB样本中的MRD测量,在新诊断,复发和PR患者中,所有FCM阴性BM样本也与FCM / IMB-FCM阴性PB样本配对,并且FCM阳性BM样本伴有IMB- FCM阳性的结果是88%的相应PB样品。CMC与疾病负担的其他临床生物标志物密切相关,包括MMC升高,β2-MG,sCrea以及DS和ISS分期,以及更严重的贫血,骨破坏和肾功能不全(P <0.05)。Logistic回归分析显示,β2-MG升高和中度至重度贫血是存在CMC的重要危险因素(P <0.05)。作为监测MRD的非侵入性“液体活检”,IMB-FCM在CMC检测中的潜力可能会补充或最小化MM患者未来的骨髓抽吸术。