Shcherbina V, Gordiienko I, Shlapatska L, Ivanivska T, Sidorenko S
Department of Molecular and Cellular Pathobiology, R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine, Kyiv 03022, Ukraine.
Department of Oncohematology, R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine, Kyiv 03022, Ukraine.
Exp Oncol. 2020 Mar;42(1):16-24. doi: 10.32471/exp-oncology.2312-8852.vol-42-no-1.14093.
Response of chronic lymphocytic leukemia (CLL) patients to classical chemoimmunotherapy that remains the main strategy in treatment of this disease is strikingly variable. This issue requires the finding of biomarkers which could predict efficiency of drug administration and choose the best treatment option for each patient individually. The aim of this study was to find out association between cell surface receptors expression levels and CLL B cells sensitivity to chemotherapeutic drugs ex vivo.
The study was performed on malignant B cells isolated from peripheral blood of primary CLL patients. Flow cytometry, qPCR, ex vivo drug sensitivity assay, and cell viability assay were used in this study.
The high CD5 expression level was linked to better bendamustine (BEN) and cyclophosphamide (CP) CLL B cells response in contrast to B cells with low CD5 expression. Sensitivity of CLL B cells to CP also could be predicted by high level of CD20 expression. Expression of CD38 and high levels of CD37 and CD40 showed CLL B cells resistance to BEN ex vivo. CLL B cells sensitivity to analyzed chemotherapeutic drugs was not dependent on CD22 expression status. The CD180 expression was detected in CLL B cells which were more susceptible to fludarabine and cyclophosphamide (FC) combinatory action. CLL B cells that coexpressed CD150 and CD180 on the cell surface were characterized by significantly decreased cell viability under fludarabine (FLU) exposure alone or FC in comparison with CD150CD180 B cells. Cell surface expression level of CD150 was not associated with CLL B cells chemosensitivity. However, high mRNA expression level of mCD150 isoform in CLL B cells was linked to their FLU sensitivity and CP resistance, while high nCD150 mRNA expression level showed resistance to FLU. Simultaneous CD150 and CD180 ligation increased FLU resistance, but BEN susceptibility of CLL B cells. CD150 and CD180 alone or in combination are involved in upregulation of CD20 cell surface expression.
Expression status of the CD5, CD20, CD37, CD38, CD40, CD150, and CD180 cell surface receptors could be used in prediction CLL B cells sensitivity to FLU, CP, BEN and FC ex vivo. Moreover, CD150 and CD180 receptors are involved in regulation of CLL B cells susceptibility to FLU and BEN. The CD150 and CD180 are positive regulators of CD20 expression that could make CD150CD180 CLL B cells more responsive to CD20-based immunotherapy.
慢性淋巴细胞白血病（CLL）患者对经典化学免疫疗法的反应差异显著，而经典化学免疫疗法仍是该疾病治疗的主要策略。这个问题需要找到能够预测药物给药效率的生物标志物，并为每个患者单独选择最佳治疗方案。本研究的目的是找出细胞表面受体表达水平与CLL B细胞体外对化疗药物敏感性之间的关联。
本研究对从原发性CLL患者外周血中分离出的恶性B细胞进行。本研究使用了流式细胞术、qPCR、体外药物敏感性测定和细胞活力测定。
与低CD5表达的B细胞相比，高CD5表达水平与苯达莫司汀（BEN）和环磷酰胺（CP）的CLL B细胞更好的反应相关。CLL B细胞对CP的敏感性也可以通过高水平的CD20表达来预测。CD38的表达以及高水平的CD37和CD40表明CLL B细胞在体外对BEN耐药。CLL B细胞对所分析化疗药物的敏感性不依赖于CD22的表达状态。在对氟达拉滨和环磷酰胺（FC）联合作用更敏感的CLL B细胞中检测到CD180表达。与CD150CD180 B细胞相比，在细胞表面共表达CD150和CD180的CLL B细胞在单独暴露于氟达拉滨（FLU）或FC时细胞活力显著降低。CD150的细胞表面表达水平与CLL B细胞的化学敏感性无关。然而，CLL B细胞中mCD150异构体的高mRNA表达水平与它们对FLU的敏感性和对CP的耐药性相关，而高nCD150 mRNA表达水平则显示对FLU耐药。同时连接CD150和CD180会增加CLL B细胞对FLU的耐药性，但增加对BEN的敏感性。单独或联合的CD150和CD180参与CD20细胞表面表达的上调。
CD5、CD20、CD37、CD38、CD40、CD150和CD180细胞表面受体的表达状态可用于预测CLL B细胞体外对FLU、CP、BEN和FC的敏感性。此外，CD150和CD180受体参与调节CLL B细胞对FLU和BEN的敏感性。CD150和CD180是CD20表达的正调节因子，这可能使CD150CD180 CLL B细胞对基于CD20的免疫疗法更敏感。
