A Nonfucosylated Anti-HER2 Antibody Augments Antibody-Dependent Cellular Cytotoxicity in Breast Cancer Patients

Suzuki, Eiji; Niwa, Rinpei; Saji, Shigehira; Muta, Mariko; Hirose, Makiko; Iida, Shigeru; Shiotsu, Yukimasa; Satoh, Mitsuo; Shitara, Kenya; Kondo, Masahide; Toi, Masakazu

doi:10.1158/1078-0432.ccr-06-1335

Cited by 99 publications

(56 citation statements)

References 36 publications

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“…S7B). In contrast to these results and in agreement with earlier studies (28,29), when PBMCs were used as effector cells, Herceptin potentiated efficient lysis of SkBr3 cells (Fig. 4B).…”

Section: E Coli Expressed Aglycosylated Trastuzumab Potentiates Thesupporting

confidence: 93%

Aglycosylated IgG variants expressed in bacteria that selectively bind FcγRI potentiate tumor cell killing by monocyte-dendritic cells

Jung¹,

Reddy²,

Kang

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

140

112

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The N-linked glycan of immunoglobulin G (IgG) is indispensable for the interaction of the Fc domain with Fcγ receptors on effector cells and the clearance of target cells via antibody dependent cell-mediated cytotoxicity (ADCC). Escherichia coli expressed, aglycosylated Fc domains bind effector FcγRs poorly and cannot elicit ADCC. Using a novel bacterial display/flow cytometric library screening system we isolated Fc variants that bind to FcγRI (CD64) with nanomolar affinity. Binding was critically dependent on amino acid substitutions (E382V, and to a lesser extent, M428I) distal to the putative FcγRI binding epitope within the CH3 domain. These mutations did not adversely affect its pH-dependent interaction with FcRn in vitro nor its serum persistence in vivo. Remarkably, the anti-Her2 IgG trastuzumab containing the E382V, M428I substitutions and expressed in E. coli exhibited highly selective binding to FcγRI but not to the other activating receptors (FcγRIIa, FcγRIIIa) nor to the inhibitory receptor, FcγRIIb. In contrast, the glycosylated version of trastuzumab (E382V, M428I) purified from HEK293T cells bound to all Fcγ receptors in a manner similar to that of clinical grade trastuzumab. E. coli-purified trastuzumab (E382V, M428I), but not glycosylated trastuzumab (E382V, M428I) or clinical grade trastuzumab, was capable of potentiating the killing of Her2 overexpressing tumor cells with dendritic cells (DCs) as effectors. These results indicate that aglycosylated IgGs can be engineered to display unique FcγR selectivity profiles that, in turn, mediate ADCC via mechanisms that are not normally displayed by glycosylated monoclonal antibodies.antibody engineering | bacterial display | bacterial expression | directed evolution | effector function

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“…S7B). In contrast to these results and in agreement with earlier studies (28,29), when PBMCs were used as effector cells, Herceptin potentiated efficient lysis of SkBr3 cells (Fig. 4B).…”

Section: E Coli Expressed Aglycosylated Trastuzumab Potentiates Thesupporting

confidence: 93%

Aglycosylated IgG variants expressed in bacteria that selectively bind FcγRI potentiate tumor cell killing by monocyte-dendritic cells

Jung¹,

Reddy²,

Kang

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

140

112

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“…Previously, there is only one report describing the efficacy of defucosylated trastuzumab using PBMCs from breast cancer patients (16). Herein, our present study is the first report indicating usefulness of defucosylated cetuximab and trastuzumab for ADCC using the PBMCs of GI-tract cancer patients.…”

Section: Discussionsupporting

confidence: 49%

“…Thus, the defucosylation technology could be one of the most powerful approaches to enhance clinical efficacy of therapeutic mAbs. There is, however, still limited information describing the clinical usefulness of the defucosylated therapeutic antibody on the ADCC, except for one report showing that the use of the defucosylated antibodies may improve the therapeutic effects of trastuzumab for breast cancer patients (16). Thus, it is necessary to draw solid conclusion for the effectiveness of the defucosylated antibody in cancer patients or immunosuppressive state.…”

Section: Introductionmentioning

confidence: 99%

Augmentation of antibody-dependent cellular cytotoxicity with defucosylated monoclonal antibodies in patients with GI-tract cancer

et al. 2017

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Abstract. Enhancement of antibody-dependent cellular cytotoxicity (ADCC) with some modalities may be a promising approach to enhance the efficacy of therapeutic monoclonal antibodies (mAbs). It has previously been demonstrated that the removal of fucose from antibody oligosaccharides (defucosylation) leads to augmentation of ADCC activity. To establish clinically relevant evidence of this procedure, the present study evaluated trastuzumab-and cetuximab-mediated ADCC by comparing defucosylated mAbs with conventional mAbs using peripheral blood mononuclear cells (PBMCs). PBMCs were isolated from 20 patients with gastrointestinal tract cancer and 10 healthy volunteers. ADCCs were measured using PBMCs as effector cells and two gastric cancer cell lines as target cells. ADCCs were significantly enhanced with defucosylated mAbs compared with conventional mAbs using PBMC from the healthy donors and patients with cancer. The results confirmed that the cetuximab-and trastuzumab-mediated ADCCs in advanced disease were impaired in comparison to those in early disease or healthy individuals. However, when the defucosylated mAbs were used instead of the conventional mAbs, the ADCC activities in the advanced cases were almost comparable with those in early disease or healthy individuals. Furthermore, the expression of ADCC associated molecules were modified toward immunosuppressive status with a mitogen-activated protein kinase inhibitor in vitro, the conventional cetuximab-and trastuzumab-mediated ADCC was downregulated, and the defucosylated mAbs overcome the downregulation of ADCC. In conclusion, defucosylated therapeutic mAbs may enhance ADCC activities in patients with cancer, which may lead to more effective anti-cancer treatments. IntroductionAlthough improvement of systemic chemotherapy has been demonstrated in gastrointestinal (GI) tract cancer including gastric and colon cancer, the overall survival rate in patients with advanced stage is still poor (1). Therefore, it would be desirable to develop molecular target therapy more efficiently in GI-tract cancer. For example, for the human epidermal growth factor receptor (EGFR) related 2 (HER2)-overexpressing gastric cancer, Trastuzumab in combination with chemotherapy vs. chemotherapy alone for treatment of HER2-positive advanced gastric cancer (ToGA) trial concluded that anti-HER2 monoclonal antibody (trastuzumab) plus chemotherapy is a standard treatment option, in which trastuzumab plus chemotherapy showed better survival in comparison to chemotherapy alone (2).It is generally accepted that trastuzumab can act on gastric cancer cells through both anti-proliferative function directly to cancer cells and antibody-dependent cellular cytotoxicity (ADCC) activity via immune cells (3,4). It has been reported that Trastuzumab-mediated ADCC can be influenced by several factors including single nucleotide polymorphisms (SNPs) in the Fc gamma receptor (FcγR) genes (5-7) or natural killer (NK) cell dysfunction (8). In fact, the SNPs can alter the FcγR binding affinity to the t...

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“…This agent showed promising evidence of activity in its phase I study and further studies are underway to determine its antitumor efficacy (Kiewe et al, 2006). In another approach, the in vitro ADCC activity of trastuzumab appears to be augmented by defucosylation, and the fucosyl-negative version of trastuzumab has been proposed for clinical testing (Suzuki et al, 2007).…”

Section: Mechanism Of Action Of Trastuzumab -Immunological Targetingmentioning

confidence: 99%

Targeting the function of the HER2 oncogene in human cancer therapeutics

2007

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A Nonfucosylated Anti-HER2 Antibody Augments Antibody-Dependent Cellular Cytotoxicity in Breast Cancer Patients

Cited by 99 publications

References 36 publications

Aglycosylated IgG variants expressed in bacteria that selectively bind FcγRI potentiate tumor cell killing by monocyte-dendritic cells

Aglycosylated IgG variants expressed in bacteria that selectively bind FcγRI potentiate tumor cell killing by monocyte-dendritic cells

Augmentation of antibody-dependent cellular cytotoxicity with defucosylated monoclonal antibodies in patients with GI-tract cancer

Targeting the function of the HER2 oncogene in human cancer therapeutics

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