FcgammaRIIIa polymorphisms and cetuximab induced cytotoxicity in squamous cell carcinoma of the head and neck - PubMed (original) (raw)

FcgammaRIIIa polymorphisms and cetuximab induced cytotoxicity in squamous cell carcinoma of the head and neck

Rodney J Taylor et al. Cancer Immunol Immunother. 2009 Jul.

Erratum in

Abstract

Purpose: The interaction of Fc fragments of antibodies with the Fcgamma receptors is an essential checkpoint in antibody-dependent cellular cytotoxicity (ADCC). Specific polymorphisms at position 158 enhance FcgammaRIIIa affinity for IgG1 and are associated with improved clinical outcome in lymphoma patients treated with IgG1 anti-CD20 antibody. The role of ADCC in the therapeutic effects of the alpha-epidermal growth factor receptor (EGFR) mAb, cetuximab, in patients with squamous cell carcinoma of the head and neck (SCCHN) is poorly defined. We employed three SCCHN cell lines to test two hypotheses: (1) SCCHN is susceptible to cetuximab-mediated ADCC, (2) efficacy of ADCC is associated with polymorphisms at position 158 of FcgammaRIIIa.

Experimental design: FcgammaRIIIa-158 polymorphisms were determined for healthy donors, and their purified NK cells were used as effector cells against three SCCHN cell lines in ADCC assays. Cytotoxicity levels were compared for each polymorphism class. Proliferation and cell cycle assays were done to examine the direct effects of cetuximab.

Results: Our results indicate that SCCHN is susceptible to cetuximab-mediated ADCC in vitro. NK cytotoxic efficiency correlates with donor 158-polymorphisms in FcgammaRIIIa. Overall cytotoxicity was greatest for individuals having a single V allele when compared to homozygous F/F individuals; the cumulative percent cytotoxicity for each polymorphism among the cell lines was 58.2% V/V, 50.6% V/F, and 26.1% F/F (P < 0.001). Additionally, the presence of a V allele correlated with superior natural cytotoxicity against NK sensitive targets.

Conclusion: These data have both prognostic and therapeutic relevance and support the design of a prospective trial to determine the influence of FcgammaRIIIa polymorphisms on the clinical outcome of patients with SCCHN treated with alpha-EGFR mAbs.

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Figures

Fig. 1

Fig. 1

Epidermal growth factor receptor expression in SCCHN tumor cell lines. SCCHN cell lines TU167, TU59, and 012SCC were stained with a α-EGFR mAb (clone ICR10) with an IgG2a isotype control and b Cetuximab with human IgG1 isotype control. Open histograms represent the isotype control. Filled histograms represent the mAbs staining. All cell lines show a comparable shift for both α-EGFR and cetuximab mAbs

Fig. 2

Fig. 2

NK FcγRIIIa polymorphisms influencing cytotoxic activity on cetuximab-treated SCCHN TU167. a NK cells purified from PBMCs of healthy donors expressing either F/F, V/F, or V/V alleles were tested for cytotoxicity against TU167 in a standard 51Cr-release assay. Before addition of effector cells, TU167 target cells were pretreated with media alone (dark filled diamond), human IgG1 isotype control at 10 μg/ml (dark filled square), or cetuximab 10 μg/ml (dark filled triangle). Data are representative of independent experiments performed in triplicates, with error bars indicating SEM. b NK cells purified from PBMCs of 17 individual donors F/F (open diamond), V/F (open square), and V/V (open triangle) alleles were tested for cytotoxicity against the TU167 SCCHN cell line using a standard 51Cr-release assay. Before addition of effector cells TU167 targets were pretreated with media alone, human IgG1 isotype control at 10 μg/ml, or cetuximab at 10 μg/ml. Donors with either FcγRIIIa V/V or FcγRIIIa V/F demonstrated significantly higher percent cytotoxicity compared to FcγRIIIa F/F, when incubated with 10 μg/ml cetuximab (P = 0.04 and 0.004, respectively)

Fig. 3

Fig. 3

FcγIIIaR V polymorphism is associated with enhanced natural cytotoxicity against K562 tumor targets. Tumor cell line K562 was subjected to direct NK cytotoxicity in the absence of antibody. Shown here, NK cells from healthy donors bearing FcγIIIaR F/F (open diamond), V/F (open square), and V/V (open triangle) alleles were tested for cytotoxicity against K562 using a standard 51Cr-release assay

Fig. 4

Fig. 4

Dose-dependent anti-proliferative activity of cetuximab measured by [3H] thymidine assays. TU159 cell line was cultured in the presence of various concentrations of cetuximab for 72 h. Proliferation was measured by thymidine incorporation. Cetuximab significantly inhibited TU159 in a dose-dependent fashion

Fig. 5

Fig. 5

Cell cycling and DNA synthesis activity of TU159 with 10 μg/ml cetuximab. Panels a, b, and c indicate representative experiments for TU159 in media alone, 10 μg/ml isotype control or 10 μg/ml cetuximab respectively. With cetuximab incubation, an increase in phase G2 + M (P5 quadrant) was observed compared to isotype control. S-phase (P6 quadrant) was decreased by at least twofold in panel c compared to panels a and b

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