title: Co-activation of STAT3 and YES-Associated Protein 1 (YAP1) Pathway in EGFR-Mutant NSCLC.
abstract: The efficacy of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) in EGFR-mutant non-small cell lung cancer (NSCLC) is limited by adaptive activation of cell survival signals. We hypothesized that both signal transducer and activator of transcription 3 (STAT3) and Src-YES-associated protein 1 (YAP1) signaling are dually activated during EGFR TKI treatment to limit therapeutic response.
  
  
  
  We used MTT and clonogenic assays, immunoblotting, and quantitative polymerase chain reaction to evaluate the efficacy of EGFR TKI alone and in combination with STAT3 and Src inhibition in three EGFR-mutant NSCLC cell lines. The Chou-Talalay method was used for the quantitative determination of drug interaction. We examined tumor growth inhibition in one EGFR-mutant NSCLC xenograft model (n = 4 mice per group). STAT3 and YAP1 expression was evaluated in tumors from 119 EGFR-mutant NSCLC patients (64 in an initial cohort and 55 in a validation cohort) by quantitative polymerase chain reaction. Kaplan-Meier and Cox regression analyses were used to assess the correlation between survival and gene expression. All statistical tests were two-sided.
  
  
  
  We discovered that lung cancer cells survive initial EGFR inhibitor treatment through activation of not only STAT3 but also Src-YAP1 signaling. Cotargeting EGFR, STAT3, and Src was synergistic in two EGFR-mutant NSCLC cell lines with a combination index of 0.59 (95% confidence interval [CI] = 0.54 to 0.63) for the PC-9 and 0.59 (95% CI = 0.54 to 0.63) for the H1975 cell line. High expression of STAT3 or YAP1 predicted worse progression-free survival (hazard ratio [HR] = 3.02, 95% CI = 1.54 to 5.93, P = .001, and HR = 2.57, 95% CI = 1.30 to 5.09, P = .007, respectively) in an initial cohort of 64 EGFR-mutant NSCLC patients treated with firstline EGFR TKIs. Similar results were observed in a validation cohort.
  
  
  
  Our study uncovers a coordinated signaling network centered on both STAT3 and Src-YAP signaling that limits targeted therapy response in lung cancer and identifies an unforeseen rational upfront polytherapy strategy to minimize residual disease and enhance clinical outcomes.
authors.affiliation: Institut d'Investigació en Ciències Germans Trias i Pujol, Badalona, Spain.
authors.forename: Imane
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authors.affiliation: Instituto Oncológico Dr. Rosell (IOR), Quirón-Dexeus University Institute, Barcelona, Spain.
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authors.affiliation: Medical Oncology, Azienda Ospedaliera Universitaria Integrata, University of Verona, Verona, Italy.
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authors.affiliation: Pangaea Biotech, Laboratory of Molecular Biology, Quirón-Dexeus University Institute, Barcelona, Spain.
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authors.affiliation: Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, China.|Laboratory of Cellular and Molecular Biology, Jiangsu Province Academy of Traditional Chinese Medicine and Jiangsu Branch of China Academy of Chinese Medical Sciences, Nanjing, China.
authors.forename: Xueting
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authors.affiliation: Shangai Pulmonary Hospital, Tongji University School of Medicine, Shangai, China.
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authors.affiliation: Pangaea Biotech, Laboratory of Molecular Biology, Quirón-Dexeus University Institute, Barcelona, Spain.
authors.forename: Carles Codony
authors.initials: CC
authors.lastname: Servat
authors.affiliation: Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, China.|Laboratory of Cellular and Molecular Biology, Jiangsu Province Academy of Traditional Chinese Medicine and Jiangsu Branch of China Academy of Chinese Medical Sciences, Nanjing, China.
authors.forename: Jie
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authors.lastname: Yang
authors.affiliation: Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, China.|Laboratory of Cellular and Molecular Biology, Jiangsu Province Academy of Traditional Chinese Medicine and Jiangsu Branch of China Academy of Chinese Medical Sciences, Nanjing, China.
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authors.affiliation: Hospital de Cruces de Barakaldo, Bizcaia, Spain.
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authors.affiliation: Hospital de Cruces de Barakaldo, Bizcaia, Spain.
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authors.affiliation: Service de Pathologie Respiratoire et d'Allergologie, CHU, Limoges France.
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authors.affiliation: Hospital Clínic, Barcelona, Spain.|Translational Genomics and Targeted Therapeutics in Solid Tumors, August Pi i Sunyer Biomedical Research Institute (IDIBAPS), Barcelona, Spain.
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authors.lastname: Reguart
authors.affiliation: Hospital La Princesa, Madrid, Spain.
authors.forename: Charo Garcia
authors.initials: CG
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authors.affiliation: Pangaea Biotech, Laboratory of Molecular Biology, Quirón-Dexeus University Institute, Barcelona, Spain.
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authors.lastname: Teixido
authors.affiliation: Hospital Puerta de Hierro, Madrid, Spain.
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authors.affiliation: Institut d'Investigació en Ciències Germans Trias i Pujol, Badalona, Spain.
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authors.affiliation: Institut d'Investigació en Ciències Germans Trias i Pujol, Badalona, Spain.
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authors.affiliation: Institut d'Investigació en Ciències Germans Trias i Pujol, Badalona, Spain.
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authors.affiliation: WV Clinical and Translational Science Institute, Morgantown, WV, USA.
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authors.affiliation: Institut Catalàd'Oncologia, Hospital Germans Trias i Pujol, Badalona, Spain.
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authors.affiliation: Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, China.|Laboratory of Cellular and Molecular Biology, Jiangsu Province Academy of Traditional Chinese Medicine and Jiangsu Branch of China Academy of Chinese Medical Sciences, Nanjing, China.
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authors.initials: TG
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authors.affiliation: Institut d'Investigació en Ciències Germans Trias i Pujol, Badalona, Spain.|Instituto Oncológico Dr. Rosell (IOR), Quirón-Dexeus University Institute, Barcelona, Spain.|Institut Catalàd'Oncologia, Hospital Germans Trias i Pujol, Badalona, Spain.|Institut Catalàd'Oncologia, Hospital Germans Trias i Pujol, Badalona, Spain.
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mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D000230:Adenocarcinoma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism*; D000077192:Adenocarcinoma of Lung; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D010750:Phosphoproteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000008:administration & dosage; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012189:Retrospective Studies; D050796:STAT3 Transcription Factor / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors; D000091102:YAP-Signaling Proteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D014157:Transcription Factors; D000091102:YAP-Signaling Proteins; C088374:YAP1 protein, human; C512478:EGFR protein, human; D066246:ErbB Receptors