Defucosylated Anti-EGFR Monoclonal Antibody 134-mG2a-f Exerts Antitumor Activities in Mouse Xenograft Models of Dog EGFR-overexpressed Cells
Defucosylated Anti-EGFR Monoclonal Antibody 134-mG2a-f Exerts Antitumor Activities in Mouse Xenograft Models of Dog EGFR-overexpressed Cells
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去岩藻糖基化抗 EGFR 单克隆抗体 134-mG2a-f 在狗 EGFR 过表达细胞的小鼠异种移植模型中发挥抗肿瘤活性
DOI：
10.1089/mab.2021.0022
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Kato Y.
中科院分区：
文献类型：
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作者：
Tateyama N;Nanamiya R;Ohishi T;Takei J;Nakamura T;Yanaka M;Hosono H;Saito M;Asano T;Tanaka T;Sano M;Kawada M;Kaneko MK;Kato Y.
表皮生长因子受体（EGFR）是I型跨膜蛋白，其是受体酪氨酸激酶的人表皮生长因子受体（HER）家族的成员。EGFR是细胞生长和分化的重要介质，并与其他HER家族成员形成同源二聚体或异源二聚体以激活下游信号级联。我们先前建立了一个抗人EGFR（hEGFR）的单克隆抗体（mAb），克隆EMab-134（小鼠IgG 1），通过免疫小鼠与hEGFR的胞外域。在本研究中，EMab-134的亚类从IgG 1转化为IgG 2a（134-mG 2a），并进一步去岩藻糖基化（134-mG 2a-f），以促进抗体依赖性细胞毒性（ADCC）。尽管134-mG 2a-f是针对hEGFR开发的，但使用流式细胞术显示其与狗EGFR（dEGFR）交叉反应。通过流式细胞术测定134-mG 2a-f对dEGFR过表达的CHO-K1（CHO/dEGFR）细胞的解离常数（KD）为3.3 × 10− 9 M，表明134-mG 2a-f对dEGFR具有高结合亲和力。体外分析显示，在靶向CHO/dEGFR细胞的实验中，134-mG 2a-f有助于高水平的ADCC和补体依赖性细胞毒性（CDC）。此外，与对对照小鼠IgG的反应中观察到的结果相比，134-mG 2a-f的体内施用显著抑制CHO/dEGFR的发展。总之，本研究的发现表明，134-mG 2a-f可用作表达dEGFR的犬癌症的治疗方案的一部分。
The epidermal growth factor receptor (EGFR) is a type I transmembrane protein, which is a member of the human epidermal growth factor receptor (HER) family of receptor tyrosine kinases. EGFR is a crucial mediator of cell growth and differentiation and forms homodimers or heterodimers with other HER family members to activate downstream signaling cascades. We previously established an anti-human EGFR (hEGFR) monoclonal antibody (mAb), clone EMab-134 (mouse IgG1), by immunizing mice with the ectodomain of hEGFR. In this study, the subclass of EMab-134 was converted from IgG1to IgG2a(134-mG2a) and further defucosylated (134-mG2a-f) to facilitate antibody-dependent cellular cytotoxicity (ADCC). Although 134-mG2a-f was developed against hEGFR, it was shown to cross-react with dog EGFR (dEGFR) using flow cytometry. The dissociation constant (KD) of 134-mG2a-f against dEGFR-overexpressed CHO-K1 (CHO/dEGFR) cells was determined by flow cytometry to be 3.3 × 10−9M, indicating that 134-mG2a-f possesses a high binding affinity to dEGFR. Analysisin vitrorevealed that 134-mG2a-f contributed to high levels of ADCC and complement-dependent cytotoxicity (CDC) in experiments targeting CHO/dEGFR cells. Furthermore, thein vivoadministration of 134-mG2a-f significantly inhibited the development of CHO/dEGFR in comparison with the results observed in response to control mouse IgG. Taken together, the findings of this study demonstrate that 134-mG2a-f could be useful as part of a therapeutic regimen for dEGFR-expressing canine cancers.