Chen Robert, Hou Jessie, Newman Edward, Kim Young, Donohue Cecile, Liu Xueli, Thomas Sandra H, Forman Stephen J, Kane Susan E
Department of Hematology and Hematopoietic Cell Transplantation, City of Hope, Duarte, California.
Department of Cancer Biology, City of Hope, Duarte, California.
Mol Cancer Ther. 2015 Jun;14(6):1376-84. doi: 10.1158/1535-7163.MCT-15-0036. Epub 2015 Apr 3.
Brentuximab vedotin (BV) is an antibody-drug conjugate that specifically delivers the potent cytotoxic drug monomethyl auristatin E (MMAE) to CD30-positive cells. BV is FDA approved for treatment of relapsed/refractory Hodgkin lymphoma and anaplastic large cell lymphoma (ALCL); however, many patients do not achieve complete remission and develop BV-resistant disease. We selected for BV-resistant Hodgkin lymphoma (L428) and ALCL (Karpas-299) cell lines using either constant (ALCL) or pulsatile (Hodgkin lymphoma) exposure to BV. We confirmed drug resistance by MTS assay and analyzed CD30 expression in resistant cells by flow cytometry, qRT-PCR, and Western blotting. We also measured drug exporter expression, MMAE resistance, and intracellular MMAE concentrations in BV-resistant cells. In addition, tissue biopsy samples from 10 Hodgkin lymphoma and 5 ALCL patients who had relapsed or progressed after BV treatment were analyzed by immunohistocytochemistry for CD30 expression. The resistant ALCL cell line, but not the Hodgkin lymphoma cell line, demonstrated downregulated CD30 expression compared with the parental cell line. In contrast, the Hodgkin lymphoma cell line, but not the ALCL cell line, exhibited MMAE resistance and increased expression of the MDR1 drug exporter compared with the parental line. For both Hodgkin lymphoma and ALCL, samples from patients relapsed/resistant on BV persistently expressed CD30 by immunohistocytochemistry. One Hodgkin lymphoma patient sample expressed MDR1 by immunohistocytochemistry. Although loss of CD30 expression is a possible mode of BV resistance in ALCL in vitro models, this has not been confirmed in patients. MMAE resistance and MDR1 expression are possible modes of BV resistance for Hodgkin lymphoma both in vitro and in patients.
本妥昔单抗（BV）是一种抗体药物偶联物，可将强效细胞毒性药物单甲基奥瑞他汀E（MMAE）特异性地递送至CD30阳性细胞。BV已获美国食品药品监督管理局（FDA）批准用于治疗复发/难治性霍奇金淋巴瘤和间变性大细胞淋巴瘤（ALCL）；然而，许多患者并未实现完全缓解，且出现了对BV耐药的疾病。我们通过持续（ALCL）或脉冲式（霍奇金淋巴瘤）暴露于BV来筛选出对BV耐药的霍奇金淋巴瘤（L428）和ALCL（Karpas - 299）细胞系。我们通过MTS试验确认了耐药性，并通过流式细胞术、定量逆转录聚合酶链反应（qRT - PCR）和蛋白质免疫印迹法分析了耐药细胞中CD30的表达。我们还测量了BV耐药细胞中药物外排转运蛋白的表达、MMAE耐药性以及细胞内MMAE浓度。此外，对10例霍奇金淋巴瘤患者和5例ALCL患者在接受BV治疗后复发或病情进展的组织活检样本进行免疫组织化学分析，以检测CD30的表达。与亲本细胞系相比，耐药的ALCL细胞系而非霍奇金淋巴瘤细胞系显示出CD30表达下调。相反，与亲本细胞系相比，霍奇金淋巴瘤细胞系而非ALCL细胞系表现出MMAE耐药性以及多药耐药蛋白1（MDR1）药物外排转运蛋白表达增加。对于霍奇金淋巴瘤和ALCL，通过免疫组织化学分析，接受BV治疗后复发/耐药患者的样本持续表达CD30。1例霍奇金淋巴瘤患者样本通过免疫组织化学表达MDR1。虽然在ALCL体外模型中CD30表达缺失是BV耐药的一种可能方式，但在患者中尚未得到证实。MMAE耐药性和MDR1表达是霍奇金淋巴瘤在体外和患者体内BV耐药的可能方式。
