Anti‑tumor effect of neratinib against lung cancer cells harboring HER2 oncogene alterations

Ogoshi,Yusuke; Shien,Kazuhiko; Yoshioka,Takahiro; Torigoe,Hidejiro; Sato,Hiroki; Sakaguchi,Masakiyo; Tomida,Shuta; Namba,Kei; Kurihara,Eisuke; Takahashi,Yuta; Suzawa,Ken; Yamamoto,Hiromasa; Soh,Junichi; Toyooka,Shinichi

doi:10.3892/ol.2019.9908

Anti‑tumor effect of neratinib against lung cancer cells harboring HER2 oncogene alterations

Authors:
- Yusuke Ogoshi
- Kazuhiko Shien
- Takahiro Yoshioka
- Hidejiro Torigoe
- Hiroki Sato
- Masakiyo Sakaguchi
- Shuta Tomida
- Kei Namba
- Eisuke Kurihara
- Yuta Takahashi
- Ken Suzawa
- Hiromasa Yamamoto
- Junichi Soh
- Shinichi Toyooka
View Affiliations

Affiliations: Department of Thoracic, Breast and Endocrinological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700‑8558, Japan, Department of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700‑8558, Japan, Department of Biobank, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700‑8558, Japan
Published online on: January 8, 2019 https://doi.org/10.3892/ol.2019.9908
Pages: 2729-2736
Copyright: © Ogoshi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Human epidermal growth factor receptor 2 (HER2) is a member of the ErbB family of receptor tyrosine kinases. Numerous studies have reported the amplification and overexpression of HER2 in several types of cancer, including non‑small cell lung cancer (NSCLC). However, the benefits of HER2‑targeted therapy have not been fully established. In the present study, the anti‑tumor effect of neratinib, an irreversible pan‑HER tyrosine kinase inhibitor (TKI), against NSCLC cells harboring HER2 alterations was investigated. The sensitivity of normal bronchial epithelial cells (BEAS‑2B) ectopically overexpressing wild‑type or mutant HER2 to neratinib was assessed. Furthermore, the anti‑tumor activity of neratinib in several NSCLC cell lines harboring HER2 alterations was determined in vitro and in vivo, and the association between their genetic alterations and sensitivity to neratinib treatment was investigated. BEAS‑2B cells ectopically overexpressing wild‑type HER2 or mutants (A775insYVMA, G776VC, G776LC, P780insGSP, V659E, G660D and S310F) exhibited constitutive autophosphorylation of HER2, as determined by western blotting. While these BEAS‑2B cells were sensitive to neratinib, they were insensitive to erlotinib, a first‑generation epidermal growth factor receptor‑TKI. Neratinib also exerted anti‑proliferative effects on HER2‑altered (H2170, Calu‑3 and H1781) NSCLC cell lines. Neratinib was also demonstrated to exert strong tumor growth inhibitory activity in mouse xenograft models using HER2‑altered lung cancer cells. The results of the present study strongly suggest that neratinib has potential as a promising therapeutic option for the treatment of HER2‑altered NSCLC.

View Figures

View References

1	Mok TS, Wu YL, Thongprasert S, Yang CH, Chu DT, Saijo N, Sunpaweravong P, Han B, Margono B, Ichinose Y, et al: Gefitinib or carboplatin-paclitaxel in pulmonary adenocarcinoma. N Engl J Med. 361:947–957. 2009. View Article : Google Scholar : PubMed/NCBI
2	Reck M, Heigener DF, Mok T, Soria JC and Rabe KF: Management of non-small-cell lung cancer: Recent developments. Lancet. 382:709–719. 2013. View Article : Google Scholar : PubMed/NCBI
3	Oxnard GR, Binder A and Janne PA: New targetable oncogenes in non-small-cell lung cancer. J Clin Oncol. 31:1097–1104. 2013. View Article : Google Scholar : PubMed/NCBI
4	Youlden DR, Cramb SM and Baade PD: The international epidemiology of lung cancer: Geographical distribution and secular trends. J Thorac Oncol. 3:819–831. 2008. View Article : Google Scholar : PubMed/NCBI
5	Moasser MM: The oncogene HER2: Its signaling and transforming functions and its role in human cancer pathogenesis. Oncogene. 26:6469–6487. 2007. View Article : Google Scholar : PubMed/NCBI
6	Yu D and Hung MC: Overexpression of ErbB2 in cancer and ErbB2-targeting strategies. Oncogene. 19:6115–6121. 2000. View Article : Google Scholar : PubMed/NCBI
7	Tan D, Deeb G, Wang J, Slocum HK, Winston J, Wiseman S, Beck A, Sait S, Anderson T and Nwogu C: HER-2/neu protein expression and gene alteration in stage I–IIIA non-small-cell lung cancer: A study of 140 cases using a combination of high throughput tissue microarray, immunohistochemistry, and fluorescent in situ hybridization. Diagn Mol Pathol. 12:201–211. 2003. View Article : Google Scholar : PubMed/NCBI
8	Pellegrini C, Falleni M, Marchetti A, Cassani B, Miozzo M, Buttitta F, Roncalli M, Coggi G and Bosari S: HER-2/Neu alterations in non-small cell lung cancer: A comprehensive evaluation by real time reverse transcription-PCR, fluorescence in situ hybridization, and immunohistochemistry. Clin Cancer Res. 9:3645–3652. 2003.PubMed/NCBI
9	Cappuzzo F, Varella-Garcia M, Shigematsu H, Domenichini I, Bartolini S, Ceresoli GL, Rossi E, Ludovini V, Gregorc V, Toschi L, et al: Increased HER2 gene copy number is associated with response to gefitinib therapy in epidermal growth factor receptor-positive non-small-cell lung cancer patients. J Clin Oncol. 23:5007–5018. 2005. View Article : Google Scholar : PubMed/NCBI
10	Swanton C, Futreal A and Eisen T: Her2-targeted therapies in non-small cell lung cancer. Clin Cancer Res. 12:4377s–4383s. 2006. View Article : Google Scholar : PubMed/NCBI
11	Stephens P, Hunter C, Bignell G, Edkins S, Davies H, Teague J, Stevens C, O'Meara S, Smith R, Parker A, et al: Lung cancer: Intragenic ERBB2 kinase mutations in tumours. Nature. 431:525–526. 2004. View Article : Google Scholar : PubMed/NCBI
12	Shigematsu H, Takahashi T, Nomura M, Majmudar K, Suzuki M, Lee H, Wistuba II, Fong KM, Toyooka S, Shimizu N, et al: Somatic mutations of the HER2 kinase domain in lung adenocarcinomas. Cancer Res. 65:1642–1646. 2005. View Article : Google Scholar : PubMed/NCBI
13	Yamamoto H, Higasa K, Sakaguchi M, Shien K, Soh J, Ichimura K, Furukawa M, Hashida S, Tsukuda K, Takigawa N, et al: Novel germline mutation in the transmembrane domain of HER2 in familial lung adenocarcinomas. J Natl Cancer Inst. 106:djt3382014. View Article : Google Scholar : PubMed/NCBI
14	Greulich H, Kaplan B, Mertins P, Chen TH, Tanaka KE, Yun CH, Zhang X, Lee SH, Cho J, Ambrogio L, et al: Functional analysis of receptor tyrosine kinase mutations in lung cancer identifies oncogenic extracellular domain mutations of ERBB2. Proc Natl Acad Sci USA. 109:14476–14481. 2012. View Article : Google Scholar : PubMed/NCBI
15	Mar N, Vredenburgh JJ and Wasser JS: Targeting HER2 in the treatment of non-small cell lung cancer. Lung Cancer. 87:220–225. 2015. View Article : Google Scholar : PubMed/NCBI
16	De G, rève J, Teugels E, Geers C, Decoster L, Galdermans D, De Mey J, Everaert H, Umelo I, In't Veld P and Schallier D: Clinical activity of afatinib (BIBW 2992) in patients with lung adenocarcinoma with mutations in the kinase domain of HER2/neu. Lung Cancer. 76:123–127. 2012. View Article : Google Scholar : PubMed/NCBI
17	Mazieres J, Peters S, Lepage B, Cortot AB, Barlesi F, Beau-Faller M, Besse B, Blons H, Mansuet-Lupo A, Urban T, et al: Lung cancer that harbors an HER2 mutation: Epidemiologic characteristics and therapeutic perspectives. J Clin Oncol. 31:1997–2003. 2013. View Article : Google Scholar : PubMed/NCBI
18	Suzawa K, Toyooka S, Sakaguchi M, Morita M, Yamamoto H, Tomida S, Ohtsuka T, Watanabe M, Hashida S, Maki Y, et al: Antitumor effect of afatinib, as a human epidermal growth factor receptor 2-targeted therapy, in lung cancers harboring HER2 oncogene alterations. Cancer Sci. 107:45–52. 2016. View Article : Google Scholar : PubMed/NCBI
19	Chan A, Delaloge S, Holmes FA, Moy B, Iwata H, Harvey VJ, Robert NJ, Silovski T, Gokmen E, von Minckwitz G, et al: Neratinib after trastuzumab-based adjuvant therapy in patients with HER2-positive breast cancer (ExteNET): A multicentre, randomised, double-blind, placebo-controlled, phase 3 trial. Lancet Oncol. 17:367–377. 2016. View Article : Google Scholar : PubMed/NCBI
20	Gandhi L, Besse B, Mazieres J, Waqar S, Cortot A, Barlesi F, Quoix E, Otterson G, Ettinger D, Horn L, et al: MA04.02 Neratinib ± temsirolimus in HER2-mutant lung cancers: An international, randomized phase II study. J Thorac Oncol. 12 Suppl:S358–S359. 2017. View Article : Google Scholar
21	Mazieres J, Barlesi F, Filleron T, Besse B, Monnet I, Beau-Faller M, Peters S, Dansin E, Früh M, Pless M, et al: Lung cancer patients with HER2 mutations treated with chemotherapy and HER2-targeted drugs: results from the European EUHER2 cohort. Ann Oncol. 27:281–286. 2016. View Article : Google Scholar : PubMed/NCBI
22	Gandhi J, Zhang J, Xie Y, Soh J, Shigematsu H, Zhang W, Yamamoto H, Peyton M, Girard L, Lockwood WW, et al: Alterations in genes of the EGFR signaling pathway and their relationship to EGFR tyrosine kinase inhibitor sensitivity in lung cancer cell lines. PLoS One. 4:e45762009. View Article : Google Scholar : PubMed/NCBI
23	Girard L, Zöchbauer-Müller S, Virmani AK, Gazdar AF and Minna JD: Genome-wide allelotyping of lung cancer identifies new regions of allelic loss, differences between small cell lung cancer and non-small cell lung cancer, and loci clustering. Cancer Res. 60:4894–4906. 2000.PubMed/NCBI
24	Sakaguchi M, Watanabe M, Kinoshita R, Kaku H, Ueki H, Futami J, Murata H, Inoue Y, Li SA, Huang P, et al: Dramatic increase in expression of a transgene by insertion of promoters downstream of the cargo gene. Mol Biotechnol. 56:621–630. 2014. View Article : Google Scholar : PubMed/NCBI
25	Shien K, Toyooka S, Ichimura K, Soh J, Furukawa M, Maki Y, Muraoka T, Tanaka N, Ueno T, Asano H, et al: Prognostic impact of cancer stem cell-related markers in non-small cell lung cancer patients treated with induction chemoradiotherapy. Lung Cancer. 77:162–167. 2012. View Article : Google Scholar : PubMed/NCBI
26	Shien K, Toyooka S, Yamamoto H, Soh J, Jida M, Thu KL, Hashida S, Maki Y, Ichihara E, Asano H, et al: Acquired resistance to EGFR inhibitors is associated with a manifestation of stem cell-like properties in cancer cells. Cancer Res. 73:3051–3061. 2013. View Article : Google Scholar : PubMed/NCBI
27	Kanda Y: Investigation of the freely available easy-to-use software ‘EZR’ for medical statistics. Bone Marrow Transplant. 48:452–458. 2013. View Article : Google Scholar : PubMed/NCBI
28	Engelman JA, Janne PA, Mermel C, Pearlberg J, Mukohara T, Fleet C, Cichowski K, Johnson BE and Cantley LC: ErbB-3 mediates phosphoinositide 3-kinase activity in gefitinib-sensitive non-small cell lung cancer cell lines. Proc Natl Acad Sc USA. 102:3788–3793. 2005. View Article : Google Scholar
29	Schwab CL, English DP, Black J, Bellone S, Lopez S, Cocco E, Bonazzoli E, Bussi B, Predolini F, Ferrari F, et al: Neratinib shows efficacy in the treatment of HER2 amplified carcinosarcoma in vitro and in vivo. Gynecol Oncol. 139:112–117. 2015. View Article : Google Scholar : PubMed/NCBI
30	Menderes G, Bonazzoli E, Bellone S, Black JD, Lopez S, Pettinella F, Masserdotti A, Zammataro L, Litkouhi B, Ratner E, et al: Efficacy of neratinib in the treatment of HER2/neu-amplified epithelial ovarian carcinoma in vitro and in vivo. Med Oncol. 34:912017. View Article : Google Scholar : PubMed/NCBI
31	Kohsaka S, Nagano M, Ueno T, Suehara Y, Hayashi T, Shimada N, Takahashi K, Suzuki K, Takamochi K, Takahashi F and Mano H: A method of high-throughput functional evaluation of EGFR gene variants of unknown significance in cancer. Sci Transl Med. 9(pii): eaan65662017. View Article : Google Scholar : PubMed/NCBI
32	Minami Y, Shimamura T, Shah K, LaFramboise T, Glatt KA, Liniker E, Borgman CL, Haringsma HJ, Feng W, Weir BA, et al: The major lung cancer-derived mutants of ERBB2 are oncogenic and are associated with sensitivity to the irreversible EGFR/ERBB2 inhibitor HKI-272. Oncogene. 26:5023–5027. 2007. View Article : Google Scholar : PubMed/NCBI
33	Tai W, Mahato R and Cheng K: The role of HER2 in cancer therapy and targeted drug delivery. J Control Release. 146:264–275. 2010. View Article : Google Scholar : PubMed/NCBI
34	Wang SE, Narasanna A, Perez-Torres M, Xiang B, Wu FY, Yang S, Carpenter G, Gazdar AF, Muthuswamy SK and Arteaga CL: HER2 kinase domain mutation results in constitutive phosphorylation and activation of HER2 and EGFR and resistance to EGFR tyrosine kinase inhibitors. Cancer Cell. 10:25–38. 2006. View Article : Google Scholar : PubMed/NCBI
35	Ou SI, Schrock AB, Bocharov EV, Klempner SJ, Haddad CK, Steinecker G, Johnson M, Gitlitz BJ, Chung J, Campregher PV, et al: HER2 transmembrane domain (TMD) mutations (V659/G660) that stabilize homo- and heterodimerization are rare oncogenic drivers in lung adenocarcinoma that respond to Afatinib. J Thorac Oncol. 12:446–457. 2017. View Article : Google Scholar : PubMed/NCBI
36	Eng J, Hsu M, Chaft JE, Kris MG, Arcila ME and Li BT: Outcomes of chemotherapies and HER2 directed therapies in advanced HER2-mutant lung cancers. Lung Cancer. 99:53–56. 2016. View Article : Google Scholar : PubMed/NCBI