Si Yingnan, Zhang Ya, Guan Jia-Shiung, Ngo Hanh Giai, Totoro Angela, Singh Ajeet Pal, Chen Kai, Xu Yuanxin, Yang Eddy S, Zhou Lufang, Liu Runhua, Liu Xiaoguang Margaret
Department of Biomedical Engineering, University of Alabama at Birmingham (UAB), 1825 University Blvd, Birmingham, AL 35294, USA.
Department of Medicine, University of Alabama at Birmingham (UAB), 703 19th Street South, Birmingham, AL 35294, USA.
Vaccines (Basel). 2021 Aug 10;9(8):882. doi: 10.3390/vaccines9080882.
Triple-negative breast cancers (TNBCs) are frequently recurrent due to the development of drug resistance post chemotherapy. Both the existing literature and our study found that surface receptor CD47 (cluster of differentiation 47) was upregulated in chemotherapy-treated TNBC cells. The goal of this study was to develop a monoclonal antibody (mAb)-based targeting strategy to treat TNBC after standard treatment. Specifically, a new mAb that targets the extracellular domain of receptor CD47 was developed using hybridoma technology and produced in fed-batch culture. Flow cytometry, confocal microscopy, and in vivo imaging system (IVIS) showed that the anti-CD47 mAb effectively targeted human and mouse TNBC cells and xenograft models with high specificity. The antibody-drug conjugate (ADC) carrying mertansine was constructed and demonstrated higher potency with reduced IC in TNBC cells than did the free drug and significantly inhibited tumor growth post gemcitabine treatment in MDA-MB-231 xenograft NSG model. Finally, whole blood analysis indicated that the anti-CD47 mAb had no general immune toxicity, flow cytometry analysis of lymph nodes revealed an increase of CD69 NK, CD11c DC, and CD4 T cells, and IHC staining showed tumoral infiltration of macrophage in the 4T1 xenograft BALB/cJ model. This study demonstrated that targeting CD47 with ADC has great potential to treat TNBCs as a targeted therapy.
三阴性乳腺癌（TNBCs）由于化疗后产生耐药性而经常复发。现有文献和我们的研究均发现，在经化疗处理的TNBC细胞中，表面受体CD47（分化簇47）上调。本研究的目的是开发一种基于单克隆抗体（mAb）的靶向策略，用于在标准治疗后治疗TNBC。具体而言，利用杂交瘤技术开发了一种靶向受体CD47胞外域的新型mAb，并在补料分批培养中生产。流式细胞术、共聚焦显微镜和体内成像系统（IVIS）显示，抗CD47 mAb能有效靶向人和小鼠TNBC细胞及异种移植模型，具有高特异性。构建了携带美登素的抗体药物偶联物（ADC），并证明其在TNBC细胞中的效力高于游离药物，IC降低，且在吉西他滨治疗后的MDA-MB-231异种移植NSG模型中显著抑制肿瘤生长。最后，全血分析表明抗CD47 mAb无一般免疫毒性，淋巴结的流式细胞术分析显示CD69 NK、CD11c DC和CD4 T细胞增加，免疫组化染色显示在4T1异种移植BALB/cJ模型中有肿瘤巨噬细胞浸润。本研究表明，用ADC靶向CD47作为一种靶向治疗方法，在治疗TNBC方面具有巨大潜力。
