A single proteolytic cleavage within the lower hinge of trastuzumab reduces immune effector function and in vivo efficacy.
A single proteolytic cleavage within the lower hinge of trastuzumab reduces immune effector function and in vivo efficacy.
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曲妥珠单抗较低铰链内的单个蛋白水解裂解可降低免疫效应子功能和体内功效。
DOI：
10.1186/bcr3240
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发表时间：
2012-08-08
期刊：
影响因子：
--
通讯作者：
An Z
中科院分区：
文献类型：
--
作者：
Fan X;Brezski RJ;Fa M;Deng H;Oberholtzer A;Gonzalez A;Dubinsky WP;Strohl WR;Jordan RE;Zhang N;An Z
最近的研究报告称，人 IgG 抗体对其下部铰链区易受特异性蛋白水解切割的影响，而铰链切割会导致 Fc 介导的效应器功能丧失。曲妥珠单抗是一种人源化IgG1治疗性单克隆抗体，用于治疗HER2过度表达的乳腺癌，其作用机制包括抑制HER2信号传导和Fc介导的抗体依赖性细胞毒性（ADCC）。本研究的目的是使用乳腺癌细胞培养方法和体内小鼠异种移植肿瘤模型来研究蛋白酶铰链切割对曲妥珠单抗疗效的潜在影响。将曲妥珠单抗与一组人基质金属蛋白酶一起孵育，并使用蛋白质印迹和质谱检测下铰链区的蛋白水解裂解。纯化单铰链裂解的曲妥珠单抗 (scIgG-T)，并评估其在体外介导 ADCC 和抑制乳腺癌细胞增殖的能力以及在小鼠异种移植肿瘤模型中的抗肿瘤功效。通过免疫组织化学检测肿瘤组织中浸润的免疫细胞。与完整的曲妥珠单抗相比，scIgG-T 保留了 HER2 抗原结合活性，并在体外抑制 HER2 介导的下游信号传导和细胞增殖。然而，scIgG-T 在体外失去了 Fc 介导的 ADCC 活性，并且在小鼠异种移植肿瘤模型中显着降低了抗肿瘤功效。免疫组织化学显示，与用完整曲妥珠单抗治疗的肿瘤组织相比，用 scIgG-T 治疗的肿瘤组织中免疫细胞浸润减少，这与体外 scIgG-T 介导的 ADCC 减少一致。曲妥珠单抗可被下铰链内的基质金属蛋白酶裂解。由于ADCC等免疫效应功能减弱，scIgG-T在体内的抗肿瘤功效显着降低。结果表明，曲妥珠单抗的较低铰链裂解可能发生在肿瘤微环境中，其中基质金属蛋白酶通常具有高水平表达，并且 scIgG-T 可能会损害其临床抗肿瘤功效。然而，需要进一步的研究来在临床环境中验证这些假设。
Recent studies reported that human IgG antibodies are susceptible to specific proteolytic cleavage in their lower hinge region, and the hinge cleavage results in a loss of Fc-mediated effector functions. Trastuzumab is a humanized IgG1 therapeutic monoclonal antibody for the treatment of HER2-overexpressing breast cancers, and its mechanisms of action consist of inhibition of HER2 signaling and Fc-mediated antibody-dependent cellular cytotoxicity (ADCC). The objective of this study is to investigate the potential effect of proteinase hinge cleavage on the efficacy of trastuzumab using both a breast cancer cell culture method and an in vivo mouse xenograft tumor model. Trastuzumab antibody was incubated with a panel of human matrix metalloproteinases, and proteolytic cleavage in the lower hinge region was detected using both western blotting and mass spectrometry. Single hinge cleaved trastuzumab (scIgG-T) was purified and evaluated for its ability to mediate ADCC and inhibition of breast cancer cell proliferation in vitro as well as anti-tumor efficacy in the mouse xenograft tumor model. Infiltrated immune cells were detected in tumor tissues by immunohistochemistry. scIgG-T retains HER2 antigen binding activity and inhibits HER2-mediated downstream signaling and cell proliferation in vitro when compared with the intact trastuzumab. However, scIgG-T lost Fc-mediated ADCC activity in vitro, and had significantly reduced anti-tumor efficacy in a mouse xenograft tumor model. Immunohistochemistry showed reduced immune cell infiltration in tumor tissues treated with scIgG-T when compared with those treated with the intact trastuzumab, which is consistent with the decreased ADCC mediated by scIgG-T in vitro. Trastuzumab can be cleaved by matrix metalloproteinases within the lower hinge. scIgG-T exhibited a significantly reduced anti-tumor efficacy in vivo due to the weakened immune effector function such as ADCC. The results suggest that the lower hinge cleavage of trastuzumab can occur in the tumor microenvironment where matrix metalloproteinases often have high levels of expression and scIgG-T might compromise its anti-tumor efficacy in the clinic. However, further studies are needed to validate these hypotheses in the clinical setting.
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影响因子：
--
作者：
Brezski RJ;Knight DM;Jordan RE
通讯作者：
Jordan RE
影响因子：
7.4
作者：
Mukherjee S;Roth MJ;Dawsey SM;Yan W;Rodriguez-Canales J;Erickson HS;Hu N;Goldstein AM;Taylor PR;Richardson AM;Tangrea MA;Chuaqui RF;Emmert-Buck MR
通讯作者：
Emmert-Buck MR
影响因子：
7.4
作者：
Beano, Alessandra;Signorino, Elena;Matera, Lina
通讯作者：
Matera, Lina
影响因子：
--
作者：
Goetz, H;Kuschel, M;Woodward, C
通讯作者：
Woodward, C
DOI：
10.1158/1078-0432.ccr-11-2294
发表时间：
2012-06-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Hurvitz SA;Betting DJ;Stern HM;Quinaux E;Stinson J;Seshagiri S;Zhao Y;Buyse M;Mackey J;Driga A;Damaraju S;Sliwkowski MX;Robert NJ;Valero V;Crown J;Falkson C;Brufsky A;Pienkowski T;Eiermann W;Martin M;Bee V;Marathe O;Slamon DJ;Timmerman JM
通讯作者：
Timmerman JM