P65BTK is a novel potential actionable target in KRAS-mutated/EGFR-wild type lung adenocarcinoma

Research output: Contribution to journalArticle

Federica Giordano, Valentina Vaira, Diego Cortinovis, Sara Bonomo, Joyce Goedmakers, Federica Brena, Annamaria Cialdella, Leonarda Ianzano, Irene Forno, Maria Grazia Cerrito, Roberto Giovannoni, Gian Luca Ferri, Ennio Tasciotti, Silve Vicent, Francesco Damarco, Silvano Bosari, Marialuisa Lavitrano, Emanuela Grassilli

Background: Lung cancer is still the main cause of cancer death worldwide despite the availability of targeted therapies and immune-checkpoint inhibitors combined with chemotherapy. Cancer cell heterogeneity and primary or acquired resistance mechanisms cause the elusive behaviour of this cancer and new biomarkers and active drugs are urgently needed to overcome these limitations. p65BTK, a novel isoform of the Bruton Tyrosine Kinase may represent a new actionable target in non-small cell lung cancer (NSCLC). Methods: p65BTK expression was evaluated by immunohistochemistry in 382 NSCLC patients with complete clinico-pathological records including smoking habit, ALK and EGFR status, and in metastatic lymph nodes of 30 NSCLC patients. NSCLC cell lines mutated for p53 and/or a component of the RAS/MAPK pathway and primary lung cancer-derived cells from Kras/Trp53 null mice were used as a preclinical model. The effects of p65BTK inhibition by BTK Tyrosine Kinase Inhibitors (TKIs) (Ibrutinib, AVL-292, RN486) and first-generation EGFR-TKIs (Gefitinib, Erlotinib) on cell viability were evaluated by MTT. The effects of BTK-TKIs on cell growth and clonogenicity were assessed by crystal violet and colony assays, respectively. Cell toxicity assays were performed to study the effect of the combination of non-toxic concentrations of BTK-TKIs with EGFR-TKIs and standard-of-care (SOC) chemotherapy (Cisplatin, Gemcitabine, Pemetrexed). Results: p65BTK was significantly over-expressed in EGFR-wild type (wt) adenocarcinomas (AdC) from non-smoker patients and its expression was also preserved at the metastatic site. p65BTK was also over-expressed in cell lines mutated for KRAS or for a component of the RAS/MAPK pathway and in tumors from Kras/Trp53 null mice. BTK-TKIs were more effective than EGFR-TKIs in decreasing cancer cell viability and significantly impaired cell proliferation and clonogenicity. Moreover, non-toxic doses of BTK-TKIs re-sensitized drug-resistant NSCLC cell lines to both target- and SOC therapy, independently from EGFR/KRAS status. Conclusions: p65BTK results as an emerging actionable target in non-smoking EGFR-wt AdC, also at advanced stages of disease. Notably, these patients are not eligible for EGFR-TKIs-based therapy due to a lack of EGFR mutation. The combination of BTK-TKIs with EGFR-TKIs is cytotoxic for EGFR-wt/KRAS-mutant/p53-null tumors and BTK-TKIs re-sensitizes drug-resistant NSCLC to SOC chemotherapy. Therefore, our data suggest that adding BTK-TKIs to SOC chemotherapy and EGFR-targeted therapy may open new avenues for clinical trials in currently untreatable NSCLC.

Original languageEnglish (US)
Article number260
JournalJournal of Experimental and Clinical Cancer Research
Volume38
Issue number1
DOIs
StatePublished - Jun 14 2019

PMID: 31200752

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P65BTK is a novel potential actionable target in KRAS-mutated/EGFR-wild type lung adenocarcinoma. / Giordano, Federica; Vaira, Valentina; Cortinovis, Diego; Bonomo, Sara; Goedmakers, Joyce; Brena, Federica; Cialdella, Annamaria; Ianzano, Leonarda; Forno, Irene; Cerrito, Maria Grazia; Giovannoni, Roberto; Ferri, Gian Luca; Tasciotti, Ennio; Vicent, Silve; Damarco, Francesco; Bosari, Silvano; Lavitrano, Marialuisa; Grassilli, Emanuela.

In: Journal of Experimental and Clinical Cancer Research, Vol. 38, No. 1, 260, 14.06.2019.

Research output: Contribution to journalArticle

Harvard

Giordano, F, Vaira, V, Cortinovis, D, Bonomo, S, Goedmakers, J, Brena, F, Cialdella, A, Ianzano, L, Forno, I, Cerrito, MG, Giovannoni, R, Ferri, GL, Tasciotti, E, Vicent, S, Damarco, F, Bosari, S, Lavitrano, M & Grassilli, E 2019, 'P65BTK is a novel potential actionable target in KRAS-mutated/EGFR-wild type lung adenocarcinoma' Journal of Experimental and Clinical Cancer Research, vol. 38, no. 1, 260. https://doi.org/10.1186/s13046-019-1199-7

APA

Giordano, F., Vaira, V., Cortinovis, D., Bonomo, S., Goedmakers, J., Brena, F., ... Grassilli, E. (2019). P65BTK is a novel potential actionable target in KRAS-mutated/EGFR-wild type lung adenocarcinoma. Journal of Experimental and Clinical Cancer Research, 38(1), [260]. https://doi.org/10.1186/s13046-019-1199-7

Vancouver

Giordano F, Vaira V, Cortinovis D, Bonomo S, Goedmakers J, Brena F et al. P65BTK is a novel potential actionable target in KRAS-mutated/EGFR-wild type lung adenocarcinoma. Journal of Experimental and Clinical Cancer Research. 2019 Jun 14;38(1). 260. https://doi.org/10.1186/s13046-019-1199-7

Author

Giordano, Federica ; Vaira, Valentina ; Cortinovis, Diego ; Bonomo, Sara ; Goedmakers, Joyce ; Brena, Federica ; Cialdella, Annamaria ; Ianzano, Leonarda ; Forno, Irene ; Cerrito, Maria Grazia ; Giovannoni, Roberto ; Ferri, Gian Luca ; Tasciotti, Ennio ; Vicent, Silve ; Damarco, Francesco ; Bosari, Silvano ; Lavitrano, Marialuisa ; Grassilli, Emanuela. / P65BTK is a novel potential actionable target in KRAS-mutated/EGFR-wild type lung adenocarcinoma. In: Journal of Experimental and Clinical Cancer Research. 2019 ; Vol. 38, No. 1.

BibTeX

@article{3233e6c895534e579bb758f9a77174de,
title = "P65BTK is a novel potential actionable target in KRAS-mutated/EGFR-wild type lung adenocarcinoma",
abstract = "Background: Lung cancer is still the main cause of cancer death worldwide despite the availability of targeted therapies and immune-checkpoint inhibitors combined with chemotherapy. Cancer cell heterogeneity and primary or acquired resistance mechanisms cause the elusive behaviour of this cancer and new biomarkers and active drugs are urgently needed to overcome these limitations. p65BTK, a novel isoform of the Bruton Tyrosine Kinase may represent a new actionable target in non-small cell lung cancer (NSCLC). Methods: p65BTK expression was evaluated by immunohistochemistry in 382 NSCLC patients with complete clinico-pathological records including smoking habit, ALK and EGFR status, and in metastatic lymph nodes of 30 NSCLC patients. NSCLC cell lines mutated for p53 and/or a component of the RAS/MAPK pathway and primary lung cancer-derived cells from Kras/Trp53 null mice were used as a preclinical model. The effects of p65BTK inhibition by BTK Tyrosine Kinase Inhibitors (TKIs) (Ibrutinib, AVL-292, RN486) and first-generation EGFR-TKIs (Gefitinib, Erlotinib) on cell viability were evaluated by MTT. The effects of BTK-TKIs on cell growth and clonogenicity were assessed by crystal violet and colony assays, respectively. Cell toxicity assays were performed to study the effect of the combination of non-toxic concentrations of BTK-TKIs with EGFR-TKIs and standard-of-care (SOC) chemotherapy (Cisplatin, Gemcitabine, Pemetrexed). Results: p65BTK was significantly over-expressed in EGFR-wild type (wt) adenocarcinomas (AdC) from non-smoker patients and its expression was also preserved at the metastatic site. p65BTK was also over-expressed in cell lines mutated for KRAS or for a component of the RAS/MAPK pathway and in tumors from Kras/Trp53 null mice. BTK-TKIs were more effective than EGFR-TKIs in decreasing cancer cell viability and significantly impaired cell proliferation and clonogenicity. Moreover, non-toxic doses of BTK-TKIs re-sensitized drug-resistant NSCLC cell lines to both target- and SOC therapy, independently from EGFR/KRAS status. Conclusions: p65BTK results as an emerging actionable target in non-smoking EGFR-wt AdC, also at advanced stages of disease. Notably, these patients are not eligible for EGFR-TKIs-based therapy due to a lack of EGFR mutation. The combination of BTK-TKIs with EGFR-TKIs is cytotoxic for EGFR-wt/KRAS-mutant/p53-null tumors and BTK-TKIs re-sensitizes drug-resistant NSCLC to SOC chemotherapy. Therefore, our data suggest that adding BTK-TKIs to SOC chemotherapy and EGFR-targeted therapy may open new avenues for clinical trials in currently untreatable NSCLC.",
keywords = "BTK inhibitors, Chemotherapy, Drug resistance, EGFR, EGFR inhibitors, NSCLC, p65BTK, Targeted therapy",
author = "Federica Giordano and Valentina Vaira and Diego Cortinovis and Sara Bonomo and Joyce Goedmakers and Federica Brena and Annamaria Cialdella and Leonarda Ianzano and Irene Forno and Cerrito, {Maria Grazia} and Roberto Giovannoni and Ferri, {Gian Luca} and Ennio Tasciotti and Silve Vicent and Francesco Damarco and Silvano Bosari and Marialuisa Lavitrano and Emanuela Grassilli",
year = "2019",
month = "6",
day = "14",
doi = "10.1186/s13046-019-1199-7",
language = "English (US)",
volume = "38",
journal = "Journal of Experimental and Clinical Cancer Research",
issn = "0392-9078",
publisher = "BioMed Central",
number = "1",

}

RIS

TY - JOUR

T1 - P65BTK is a novel potential actionable target in KRAS-mutated/EGFR-wild type lung adenocarcinoma

AU - Giordano, Federica

AU - Vaira, Valentina

AU - Cortinovis, Diego

AU - Bonomo, Sara

AU - Goedmakers, Joyce

AU - Brena, Federica

AU - Cialdella, Annamaria

AU - Ianzano, Leonarda

AU - Forno, Irene

AU - Cerrito, Maria Grazia

AU - Giovannoni, Roberto

AU - Ferri, Gian Luca

AU - Tasciotti, Ennio

AU - Vicent, Silve

AU - Damarco, Francesco

AU - Bosari, Silvano

AU - Lavitrano, Marialuisa

AU - Grassilli, Emanuela

PY - 2019/6/14

Y1 - 2019/6/14

N2 - Background: Lung cancer is still the main cause of cancer death worldwide despite the availability of targeted therapies and immune-checkpoint inhibitors combined with chemotherapy. Cancer cell heterogeneity and primary or acquired resistance mechanisms cause the elusive behaviour of this cancer and new biomarkers and active drugs are urgently needed to overcome these limitations. p65BTK, a novel isoform of the Bruton Tyrosine Kinase may represent a new actionable target in non-small cell lung cancer (NSCLC). Methods: p65BTK expression was evaluated by immunohistochemistry in 382 NSCLC patients with complete clinico-pathological records including smoking habit, ALK and EGFR status, and in metastatic lymph nodes of 30 NSCLC patients. NSCLC cell lines mutated for p53 and/or a component of the RAS/MAPK pathway and primary lung cancer-derived cells from Kras/Trp53 null mice were used as a preclinical model. The effects of p65BTK inhibition by BTK Tyrosine Kinase Inhibitors (TKIs) (Ibrutinib, AVL-292, RN486) and first-generation EGFR-TKIs (Gefitinib, Erlotinib) on cell viability were evaluated by MTT. The effects of BTK-TKIs on cell growth and clonogenicity were assessed by crystal violet and colony assays, respectively. Cell toxicity assays were performed to study the effect of the combination of non-toxic concentrations of BTK-TKIs with EGFR-TKIs and standard-of-care (SOC) chemotherapy (Cisplatin, Gemcitabine, Pemetrexed). Results: p65BTK was significantly over-expressed in EGFR-wild type (wt) adenocarcinomas (AdC) from non-smoker patients and its expression was also preserved at the metastatic site. p65BTK was also over-expressed in cell lines mutated for KRAS or for a component of the RAS/MAPK pathway and in tumors from Kras/Trp53 null mice. BTK-TKIs were more effective than EGFR-TKIs in decreasing cancer cell viability and significantly impaired cell proliferation and clonogenicity. Moreover, non-toxic doses of BTK-TKIs re-sensitized drug-resistant NSCLC cell lines to both target- and SOC therapy, independently from EGFR/KRAS status. Conclusions: p65BTK results as an emerging actionable target in non-smoking EGFR-wt AdC, also at advanced stages of disease. Notably, these patients are not eligible for EGFR-TKIs-based therapy due to a lack of EGFR mutation. The combination of BTK-TKIs with EGFR-TKIs is cytotoxic for EGFR-wt/KRAS-mutant/p53-null tumors and BTK-TKIs re-sensitizes drug-resistant NSCLC to SOC chemotherapy. Therefore, our data suggest that adding BTK-TKIs to SOC chemotherapy and EGFR-targeted therapy may open new avenues for clinical trials in currently untreatable NSCLC.

AB - Background: Lung cancer is still the main cause of cancer death worldwide despite the availability of targeted therapies and immune-checkpoint inhibitors combined with chemotherapy. Cancer cell heterogeneity and primary or acquired resistance mechanisms cause the elusive behaviour of this cancer and new biomarkers and active drugs are urgently needed to overcome these limitations. p65BTK, a novel isoform of the Bruton Tyrosine Kinase may represent a new actionable target in non-small cell lung cancer (NSCLC). Methods: p65BTK expression was evaluated by immunohistochemistry in 382 NSCLC patients with complete clinico-pathological records including smoking habit, ALK and EGFR status, and in metastatic lymph nodes of 30 NSCLC patients. NSCLC cell lines mutated for p53 and/or a component of the RAS/MAPK pathway and primary lung cancer-derived cells from Kras/Trp53 null mice were used as a preclinical model. The effects of p65BTK inhibition by BTK Tyrosine Kinase Inhibitors (TKIs) (Ibrutinib, AVL-292, RN486) and first-generation EGFR-TKIs (Gefitinib, Erlotinib) on cell viability were evaluated by MTT. The effects of BTK-TKIs on cell growth and clonogenicity were assessed by crystal violet and colony assays, respectively. Cell toxicity assays were performed to study the effect of the combination of non-toxic concentrations of BTK-TKIs with EGFR-TKIs and standard-of-care (SOC) chemotherapy (Cisplatin, Gemcitabine, Pemetrexed). Results: p65BTK was significantly over-expressed in EGFR-wild type (wt) adenocarcinomas (AdC) from non-smoker patients and its expression was also preserved at the metastatic site. p65BTK was also over-expressed in cell lines mutated for KRAS or for a component of the RAS/MAPK pathway and in tumors from Kras/Trp53 null mice. BTK-TKIs were more effective than EGFR-TKIs in decreasing cancer cell viability and significantly impaired cell proliferation and clonogenicity. Moreover, non-toxic doses of BTK-TKIs re-sensitized drug-resistant NSCLC cell lines to both target- and SOC therapy, independently from EGFR/KRAS status. Conclusions: p65BTK results as an emerging actionable target in non-smoking EGFR-wt AdC, also at advanced stages of disease. Notably, these patients are not eligible for EGFR-TKIs-based therapy due to a lack of EGFR mutation. The combination of BTK-TKIs with EGFR-TKIs is cytotoxic for EGFR-wt/KRAS-mutant/p53-null tumors and BTK-TKIs re-sensitizes drug-resistant NSCLC to SOC chemotherapy. Therefore, our data suggest that adding BTK-TKIs to SOC chemotherapy and EGFR-targeted therapy may open new avenues for clinical trials in currently untreatable NSCLC.

KW - BTK inhibitors

KW - Chemotherapy

KW - Drug resistance

KW - EGFR

KW - EGFR inhibitors

KW - NSCLC

KW - p65BTK

KW - Targeted therapy

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UR - http://www.scopus.com/inward/citedby.url?scp=85067273485&partnerID=8YFLogxK

U2 - 10.1186/s13046-019-1199-7

DO - 10.1186/s13046-019-1199-7

M3 - Article

VL - 38

JO - Journal of Experimental and Clinical Cancer Research

T2 - Journal of Experimental and Clinical Cancer Research

JF - Journal of Experimental and Clinical Cancer Research

SN - 0392-9078

IS - 1

M1 - 260

ER -

ID: 49921213