Development of a recombinant anti-VEGFR2-EPCAM bispecific antibody to improve antiangiogenic efficiency
Development of a recombinant anti-VEGFR2-EPCAM bispecific antibody to improve antiangiogenic efficiency
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DOI：
10.1016/j.yexcr.2021.112685
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发表时间：
2021-06-16
影响因子：
3.7
通讯作者：
Farahmand, Leila
中科院分区：
文献类型：
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作者：
Barzaman, Khadijeh;Samadi, Mitra;Farahmand, Leila
肿瘤的进展和转移，特别是在浸润性癌症（如三阴性乳腺癌[TNBC]）中，依赖于血管生成，其中血管上皮生长因子（VEGF）/血管上皮生长因子受体[1]具有决定性作用，其次是癌细胞的转移性扩散。虽然一些研究表明抗VEGFR 2/VEGF单克隆抗体在临床上显示出良好的结果，但这种方法并不有效，需要进一步研究以提高癌症治疗的质量。此外，上皮细胞粘附分子（EpCAM）在各种癌症（例如，浸润性乳腺癌）中的表达增加有助于血管生成，促进肿瘤细胞迁移到身体的其他部位。因此，我们研究的主要目标是靶向VEGFR 2或EpCAM作为乳腺癌血管生成进展中的关键参与者。关于癌症治疗，与靶向一种以上抗原或受体的单克隆抗体相比，双特异性抗体的生产更容易且更具成本效益;为此，我们通过双特异性抗体生产了靶向表达EpCAM和VEGFR 2的细胞的重组抗体，以减少肿瘤细胞的增殖和转移。在E. Western blot分析证实表达产物的正确性，流式细胞仪检测表达产物与MDA-MB-231和MCF-7细胞株上VEGFR 2和EPCAM的结合活性。用MTT法和细胞凋亡实验检测抗体对肿瘤细胞增殖的抑制作用。随后，数据表明，在乳腺癌细胞系中，通过双特异性抗体抑制了迁移、侵袭和血管生成。此外，细胞因子分析表明，双特异性抗体可以调节白细胞介素8（IL-8）和IL-6作为乳腺癌血管生成进展中的关键介质。因此，我们的双特异性抗体可以被认为是减少TNBC中血管生成的有前景的候选工具。
Tumor progression and metastasis, especially in invasive cancers (such as triple-negative breast cancer [TNBC]), depend on angiogenesis, in which vascular epithelial growth factor (VEGF)/vascular epithelial growth factor receptor [1] has a decisive role, followed by the metastatic spread of cancer cells. Although some studies have shown that anti-VEGFR2/VEGF monoclonal antibodies demonstrated favorable results in the clinic, this approach is not efficient, and further investigations are needed to improve the quality of cancer treatment. Besides, the increased expression of epithelial cell adhesion molecule (EpCAM) in various cancers, for instance, invasive breast cancer, contributes to angiogenesis, facilitating the migration of tumor cells to other parts of the body. Thus, the main goal of our study was to target either VEGFR2 or EpCAM as pivotal players in the progression of angiogenesis in breast cancer. Regarding cancer therapy, the production of bispecific antibodies is easier and more cost-effective compared to monoclonal antibodies, targeting more than one antigen or receptor; for this reason, we produced a recombinant antibody to target cells expressing EpCAM and VEGFR2 via a bispecific antibody to decrease the proliferation and metastasis of tumor cells. Following the cloning and expression of our desired anti-VEGFR2/EPCAM sequence in E. coli, the accuracy of the expression was confirmed by Western blot analysis, and its binding activities to VEGFR2 and EPCAM on MDA-MB-231 and MCF-7 cell lines were respectively indicated by flow cytometry. Then, its anti-proliferative potential was indicated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and apoptosis assay to evaluate inhibitory effects of the antibody on tumor cells. Subsequently, the data indicated that migration, invasion, and angiogenesis were inhibited in breast cancer cell lines via the bispecific antibody. Furthermore, cytokine analysis indicated that the bispecific antibody could moderate interleukin 8 (IL-8) and IL-6 as key mediators in angiogenesis progression in breast cancer. Thus, our bispecific antibody could be considered as a promising candidate tool to decrease angiogenesis in TNBC.