Analysis of in vitro ADCC and clinical response to trastuzumab

Possible relevance of FcγRIIIA/FcγRIIA gene polymorphisms and HER-2 expression levels on breast cancer cell lines

Silvia Boero, Anna Morabito, Barbara Banelli, Barbara Cardinali, Beatrice Dozin, Gianluigi Lunardi, Patrizia Piccioli, Sonia Lastraioli, Roberta Carosio, Sandra Salvi, Alessia Levaggi, Francesca Poggio, Alessia D'Alonzo, Massimo Romani, Lucia Mastro, Alessandro Poggi, Maria Pia Pistillo

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Background: Trastuzumab is a humanized monoclonal antibody (mAb) currently used for the treatment of breast cancer (BC) patients with HER-2 overexpressing tumor subtype. Previous data reported the involvement of FcγRIIIA/IIA gene polymorphisms and/or antibody-dependent cellular cytotoxicity (ADCC) in the therapeutic efficacy of trastuzumab, although results on these issues are still controversial. This study was aimed to evaluate in vitro the functional relationships among FcγRIIIA/IIA polymorphisms, ADCC intensity and HER-2 expression on tumor target cells and to correlate them with response to trastuzumab. Patients and methods: Twenty-five patients with HER-2 overexpressing BC, receiving trastuzumabin a neoadjuvant (NEO) or metastatic (MTS) setting, were genotyped for the FcγRIIIA 158V>F and FcγRIIA 131H>R polymorphisms by a newly developed pyrosequencing assay and by multiplex Tetra-primer-ARMS PCR, respectively. Trastuzumab-mediated ADCC of patients' peripheral blood mononuclear cells (PBMCs) was evaluated prior to therapy and measured by 51Chromium release using as targets three human BC cell lines showing different levels of reactivity with trastuzumab. Results: We found that the FcγRIIIA 158F and/or the FcγRIIA 131R variants, commonly reported as unfavorable in BC, may actually behave as ADCC favorable genotypes, in both the NEO (P ranging from 0.009 to 0.039 and from 0.007 to 0.047, respectively) and MTS (P ranging from 0.009 to 0.032 and P=0.034, respectively) patients. The ADCC intensity was affected by different levels of trastuzumab reactivity with BC target cells. In this context, the MCF-7 cell line, showing the lowest reactivity with trastuzumab, resulted the most suitable cell line for evaluating ADCC and response to trastuzumab. Indeed, we found a statistically significant correlation between an increased frequency of patients showing ADCC of MCF-7 and complete response to trastuzumab in the NEO setting (P=0.006). Conclusions: Although this study was performed in a limited number of patients, it would indicate a correlation of FcγR gene polymorphisms to the ADCC extent in combination with the HER-2 expression levels on tumor target cells in BC patients. However, to confirm our findings further experimental evidences obtained from a larger cohort of BC patients are mandatory.

Original languageEnglish
Article number324
JournalJournal of Translational Medicine
Volume13
Issue number1
DOIs
Publication statusPublished - Oct 8 2015

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erbB-2 Genes
Cytotoxicity
Polymorphism
Genes
Cells
Breast Neoplasms
Cell Line
Antibodies
Tumors
Antibodies, Monoclonal, Humanized
Trastuzumab
In Vitro Techniques
Neoplasms
MCF-7 Cells
Assays
Blood Cells
Blood
Therapeutics
Genotype
Polymerase Chain Reaction

Keywords

  • ADCC
  • Breast cancer patients
  • FcγR single nucleotide polymorphisms
  • Trastuzumab

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Analysis of in vitro ADCC and clinical response to trastuzumab : Possible relevance of FcγRIIIA/FcγRIIA gene polymorphisms and HER-2 expression levels on breast cancer cell lines. / Boero, Silvia; Morabito, Anna; Banelli, Barbara; Cardinali, Barbara; Dozin, Beatrice; Lunardi, Gianluigi; Piccioli, Patrizia; Lastraioli, Sonia; Carosio, Roberta; Salvi, Sandra; Levaggi, Alessia; Poggio, Francesca; D'Alonzo, Alessia; Romani, Massimo; Mastro, Lucia; Poggi, Alessandro; Pistillo, Maria Pia.

In: Journal of Translational Medicine, Vol. 13, No. 1, 324, 08.10.2015.

Research output: Contribution to journalArticle

Boero, Silvia ; Morabito, Anna ; Banelli, Barbara ; Cardinali, Barbara ; Dozin, Beatrice ; Lunardi, Gianluigi ; Piccioli, Patrizia ; Lastraioli, Sonia ; Carosio, Roberta ; Salvi, Sandra ; Levaggi, Alessia ; Poggio, Francesca ; D'Alonzo, Alessia ; Romani, Massimo ; Mastro, Lucia ; Poggi, Alessandro ; Pistillo, Maria Pia. / Analysis of in vitro ADCC and clinical response to trastuzumab : Possible relevance of FcγRIIIA/FcγRIIA gene polymorphisms and HER-2 expression levels on breast cancer cell lines. In: Journal of Translational Medicine. 2015 ; Vol. 13, No. 1.
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abstract = "Background: Trastuzumab is a humanized monoclonal antibody (mAb) currently used for the treatment of breast cancer (BC) patients with HER-2 overexpressing tumor subtype. Previous data reported the involvement of FcγRIIIA/IIA gene polymorphisms and/or antibody-dependent cellular cytotoxicity (ADCC) in the therapeutic efficacy of trastuzumab, although results on these issues are still controversial. This study was aimed to evaluate in vitro the functional relationships among FcγRIIIA/IIA polymorphisms, ADCC intensity and HER-2 expression on tumor target cells and to correlate them with response to trastuzumab. Patients and methods: Twenty-five patients with HER-2 overexpressing BC, receiving trastuzumabin a neoadjuvant (NEO) or metastatic (MTS) setting, were genotyped for the FcγRIIIA 158V>F and FcγRIIA 131H>R polymorphisms by a newly developed pyrosequencing assay and by multiplex Tetra-primer-ARMS PCR, respectively. Trastuzumab-mediated ADCC of patients' peripheral blood mononuclear cells (PBMCs) was evaluated prior to therapy and measured by 51Chromium release using as targets three human BC cell lines showing different levels of reactivity with trastuzumab. Results: We found that the FcγRIIIA 158F and/or the FcγRIIA 131R variants, commonly reported as unfavorable in BC, may actually behave as ADCC favorable genotypes, in both the NEO (P ranging from 0.009 to 0.039 and from 0.007 to 0.047, respectively) and MTS (P ranging from 0.009 to 0.032 and P=0.034, respectively) patients. The ADCC intensity was affected by different levels of trastuzumab reactivity with BC target cells. In this context, the MCF-7 cell line, showing the lowest reactivity with trastuzumab, resulted the most suitable cell line for evaluating ADCC and response to trastuzumab. Indeed, we found a statistically significant correlation between an increased frequency of patients showing ADCC of MCF-7 and complete response to trastuzumab in the NEO setting (P=0.006). Conclusions: Although this study was performed in a limited number of patients, it would indicate a correlation of FcγR gene polymorphisms to the ADCC extent in combination with the HER-2 expression levels on tumor target cells in BC patients. However, to confirm our findings further experimental evidences obtained from a larger cohort of BC patients are mandatory.",
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TY - JOUR

T1 - Analysis of in vitro ADCC and clinical response to trastuzumab

T2 - Possible relevance of FcγRIIIA/FcγRIIA gene polymorphisms and HER-2 expression levels on breast cancer cell lines

AU - Boero, Silvia

AU - Morabito, Anna

AU - Banelli, Barbara

AU - Cardinali, Barbara

AU - Dozin, Beatrice

AU - Lunardi, Gianluigi

AU - Piccioli, Patrizia

AU - Lastraioli, Sonia

AU - Carosio, Roberta

AU - Salvi, Sandra

AU - Levaggi, Alessia

AU - Poggio, Francesca

AU - D'Alonzo, Alessia

AU - Romani, Massimo

AU - Mastro, Lucia

AU - Poggi, Alessandro

AU - Pistillo, Maria Pia

PY - 2015/10/8

Y1 - 2015/10/8

N2 - Background: Trastuzumab is a humanized monoclonal antibody (mAb) currently used for the treatment of breast cancer (BC) patients with HER-2 overexpressing tumor subtype. Previous data reported the involvement of FcγRIIIA/IIA gene polymorphisms and/or antibody-dependent cellular cytotoxicity (ADCC) in the therapeutic efficacy of trastuzumab, although results on these issues are still controversial. This study was aimed to evaluate in vitro the functional relationships among FcγRIIIA/IIA polymorphisms, ADCC intensity and HER-2 expression on tumor target cells and to correlate them with response to trastuzumab. Patients and methods: Twenty-five patients with HER-2 overexpressing BC, receiving trastuzumabin a neoadjuvant (NEO) or metastatic (MTS) setting, were genotyped for the FcγRIIIA 158V>F and FcγRIIA 131H>R polymorphisms by a newly developed pyrosequencing assay and by multiplex Tetra-primer-ARMS PCR, respectively. Trastuzumab-mediated ADCC of patients' peripheral blood mononuclear cells (PBMCs) was evaluated prior to therapy and measured by 51Chromium release using as targets three human BC cell lines showing different levels of reactivity with trastuzumab. Results: We found that the FcγRIIIA 158F and/or the FcγRIIA 131R variants, commonly reported as unfavorable in BC, may actually behave as ADCC favorable genotypes, in both the NEO (P ranging from 0.009 to 0.039 and from 0.007 to 0.047, respectively) and MTS (P ranging from 0.009 to 0.032 and P=0.034, respectively) patients. The ADCC intensity was affected by different levels of trastuzumab reactivity with BC target cells. In this context, the MCF-7 cell line, showing the lowest reactivity with trastuzumab, resulted the most suitable cell line for evaluating ADCC and response to trastuzumab. Indeed, we found a statistically significant correlation between an increased frequency of patients showing ADCC of MCF-7 and complete response to trastuzumab in the NEO setting (P=0.006). Conclusions: Although this study was performed in a limited number of patients, it would indicate a correlation of FcγR gene polymorphisms to the ADCC extent in combination with the HER-2 expression levels on tumor target cells in BC patients. However, to confirm our findings further experimental evidences obtained from a larger cohort of BC patients are mandatory.

AB - Background: Trastuzumab is a humanized monoclonal antibody (mAb) currently used for the treatment of breast cancer (BC) patients with HER-2 overexpressing tumor subtype. Previous data reported the involvement of FcγRIIIA/IIA gene polymorphisms and/or antibody-dependent cellular cytotoxicity (ADCC) in the therapeutic efficacy of trastuzumab, although results on these issues are still controversial. This study was aimed to evaluate in vitro the functional relationships among FcγRIIIA/IIA polymorphisms, ADCC intensity and HER-2 expression on tumor target cells and to correlate them with response to trastuzumab. Patients and methods: Twenty-five patients with HER-2 overexpressing BC, receiving trastuzumabin a neoadjuvant (NEO) or metastatic (MTS) setting, were genotyped for the FcγRIIIA 158V>F and FcγRIIA 131H>R polymorphisms by a newly developed pyrosequencing assay and by multiplex Tetra-primer-ARMS PCR, respectively. Trastuzumab-mediated ADCC of patients' peripheral blood mononuclear cells (PBMCs) was evaluated prior to therapy and measured by 51Chromium release using as targets three human BC cell lines showing different levels of reactivity with trastuzumab. Results: We found that the FcγRIIIA 158F and/or the FcγRIIA 131R variants, commonly reported as unfavorable in BC, may actually behave as ADCC favorable genotypes, in both the NEO (P ranging from 0.009 to 0.039 and from 0.007 to 0.047, respectively) and MTS (P ranging from 0.009 to 0.032 and P=0.034, respectively) patients. The ADCC intensity was affected by different levels of trastuzumab reactivity with BC target cells. In this context, the MCF-7 cell line, showing the lowest reactivity with trastuzumab, resulted the most suitable cell line for evaluating ADCC and response to trastuzumab. Indeed, we found a statistically significant correlation between an increased frequency of patients showing ADCC of MCF-7 and complete response to trastuzumab in the NEO setting (P=0.006). Conclusions: Although this study was performed in a limited number of patients, it would indicate a correlation of FcγR gene polymorphisms to the ADCC extent in combination with the HER-2 expression levels on tumor target cells in BC patients. However, to confirm our findings further experimental evidences obtained from a larger cohort of BC patients are mandatory.

KW - ADCC

KW - Breast cancer patients

KW - FcγR single nucleotide polymorphisms

KW - Trastuzumab

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DO - 10.1186/s12967-015-0680-0

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