title: Cytotoxicity of allitinib, an irreversible anti-EGFR agent, in a large panel of human cancer-derived cell lines: KRAS mutation status as a predictive biomarker.
abstract: BACKGROUND
  The epidermal growth factor receptor (EGFR) is a member of the HER family of growth factors that activates several intracellular signaling pathways promoting proliferation and survival. EGFR over-expression is frequently associated with gene mutation or amplification, thereby constituting a major target for molecular therapies. Recently, a new generation of EGFR inhibitors has been developed with pan-HER properties and irreversible actions. Allitinib® (AST1306) is an orally active, highly selective irreversible inhibitor of the HER family of receptor tyrosine kinases with promising efficacies. In the present study we aimed to investigate the cytotoxicity of allitinib in a large panel of human cancer-derived cell lines and to correlate its efficacy to the mutational status of the EGFR, KRAS, BRAF, PI3KCA and PTEN genes. In addition, we aimed to evaluate the functional role of KRAS mutations in the response to this new inhibitor.
  
  
  RESULTS
  In total 76 different cancer-derived cell lines, representing 11 distinct histological types, were analyzed and classified into three groups: highly sensitive (HS), moderately sensitive (MS) and resistant (R). We found that 28 (36.8 %) cancer-derived cell lines exhibited a HS phenotype, 19 (25.0 %) a MS phenotype and 29 (38.1 %) a R phenotype. Allitinib showed a stronger cytotoxicity in head and neck, esophageal, melanoma and lung cancer-derived cell lines. We found that KRAS mutations were significantly associated with the R phenotype. To substantiate these results, an allitinib-sensitive lung cancer-derived cell line (H292) was transfected with plasmids carrying the two most common activating KRAS mutations (p.G12D and p.G12S). We found that both mutations reverted the allitinib-sensitive phenotype in these cells.
  
  
  CONCLUSIONS
  The current study represents the largest in vitro assessment of allitinib cytotoxicity performed to date. Through this study, we identified cancer types that could potentially benefit from this drug. Additionally, our findings suggest that prevalent KRAS mutations constitute potential predictive biomarkers for allitinib response.
authors.affiliation: Molecular Oncology Research Center, Barretos Cancer Hospital, Rua Antenor Duarte Villela, 1331, CEP 14784 400, Barretos, São Paulo, Brazil.
authors.forename: Renato José
authors.initials: RJ
authors.lastname: Silva-Oliveira
authors.affiliation: Molecular Oncology Research Center, Barretos Cancer Hospital, Rua Antenor Duarte Villela, 1331, CEP 14784 400, Barretos, São Paulo, Brazil.
authors.forename: Viviane Aline Oliveira
authors.initials: VA
authors.lastname: Silva
authors.affiliation: Molecular Oncology Research Center, Barretos Cancer Hospital, Rua Antenor Duarte Villela, 1331, CEP 14784 400, Barretos, São Paulo, Brazil.|Life and Health Sciences Research Institute (ICVS), Health Sciences School, University of Minho, Braga, Portugal.|ICVS/3B's-PT Government Associate Laboratory, Braga, Guimarães, Portugal.
authors.forename: Olga
authors.initials: O
authors.lastname: Martinho
authors.affiliation: Molecular Oncology Research Center, Barretos Cancer Hospital, Rua Antenor Duarte Villela, 1331, CEP 14784 400, Barretos, São Paulo, Brazil.
authors.forename: Adriana
authors.initials: A
authors.lastname: Cruvinel-Carloni
authors.affiliation: Molecular Oncology Research Center, Barretos Cancer Hospital, Rua Antenor Duarte Villela, 1331, CEP 14784 400, Barretos, São Paulo, Brazil.
authors.forename: Matias Eliseo
authors.initials: ME
authors.lastname: Melendez
authors.affiliation: Molecular Oncology Research Center, Barretos Cancer Hospital, Rua Antenor Duarte Villela, 1331, CEP 14784 400, Barretos, São Paulo, Brazil.
authors.forename: Marcela Nunes
authors.initials: MN
authors.lastname: Rosa
authors.affiliation: Molecular Oncology Research Center, Barretos Cancer Hospital, Rua Antenor Duarte Villela, 1331, CEP 14784 400, Barretos, São Paulo, Brazil.
authors.forename: Flávia Escremim
authors.initials: FE
authors.lastname: de Paula
authors.affiliation: Molecular Oncology Research Center, Barretos Cancer Hospital, Rua Antenor Duarte Villela, 1331, CEP 14784 400, Barretos, São Paulo, Brazil.|Department of Medical Oncology, Barretos Cancer Hospital, Barretos, São Paulo, Brazil.
authors.forename: Luciano
authors.initials: L
authors.lastname: de Souza Viana
authors.affiliation: Molecular Oncology Research Center, Barretos Cancer Hospital, Rua Antenor Duarte Villela, 1331, CEP 14784 400, Barretos, São Paulo, Brazil.
authors.forename: André Lopes
authors.initials: AL
authors.lastname: Carvalho
authors.affiliation: Molecular Oncology Research Center, Barretos Cancer Hospital, Rua Antenor Duarte Villela, 1331, CEP 14784 400, Barretos, São Paulo, Brazil. ruireis.hcb@gmail.com.|Life and Health Sciences Research Institute (ICVS), Health Sciences School, University of Minho, Braga, Portugal. ruireis.hcb@gmail.com.|ICVS/3B's-PT Government Associate Laboratory, Braga, Guimarães, Portugal. ruireis.hcb@gmail.com.
authors.forename: Rui Manuel
authors.initials: RM
authors.lastname: Reis
mesh_terms: D000178:Acrylamides / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004252:DNA Mutational Analysis; D004354:Drug Screening Assays, Antitumor; D066246:ErbB Receptors / Q000037:antagonists & inhibitors; D006801:Humans; D009154:Mutation; D009369:Neoplasms / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics*; D011799:Quinazolines / Q000494:pharmacology*
chemical_list: C568037:AST 1306; D000178:Acrylamides; C117307:KRAS protein, human; D011799:Quinazolines; C512478:EGFR protein, human; D066246:ErbB Receptors; D016283:Proto-Oncogene Proteins p21(ras)
keywords: Allitinib; EGFR inhibitor; In vitro screening; KRAS mutation