Identification and Characterization of ALK Kinase Splicing Isoforms in Non-Small-Cell Lung Cancer

Lorena Lobo De Figueiredo-Pontes, Daisy Wing Sze Wong, Vicky Pui Chi Tin, Lap Ping Chung, Hiroyuki Yasuda, Norihiro Yamaguchi, Sohei Nakayama, Pasi Antero Jänne, Maria Pik Wong, Susumu Soeda Kobayashi, Daniel Botelho Costa

Research output: Contribution to journalArticle

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Abstract

INTRODUCTION:: Anaplastic lymphoma kinase (ALK) rearrangements are present in an important subset of non-small-cell lung cancer (NSCLC) and predict for response to the tyrosine kinase inhibitor crizotinib. In this study, we evaluated the yet unknown frequency and functional role of ALK splicing isoforms in NSCLC. METHODS:: We analyzed 270 cases of NSCLC for ALK kinase domain splicing aberrations and in addition generated constructs with full-length echinoderm microtubule-associated protein-like 4 (EML4)-ALK (E13;A20) and a splicing isoform. RESULTS:: Splicing isoforms of the kinase domain of ALK - including complete skipping of exon 23 (ALKdel23, ALK p.I1171fs*42) and exon 27 (ALKdel27, ALK p.T1312fs*0) - were identified in 11.1% (30 of 270 cases) of NSCLC, and these changes coexisted with ALK rearrangements, KRAS mutations, and EGFR mutations. ALK splicing isoforms were observed with full-length EML4-ALK in crizotinib-naive and treated NSCLCs. ALK T1312fs*0 was unable to render cells solely dependent on ALK signaling. Unlike EML4-ALK and EML4-ALK p.L1196M, EML4-ALK T1312fs*0 did not autophosphorylate ALK or other phosphotyrosine sites. Coexpression of equal amounts of EML4-ALK T1312fs*0 and EML4-ALK did not result in resistance to crizotinib, whereas coexpression of EML4-ALK L1196M with EML4-ALK resulted in resistance to inhibition of ALK by crizotinib. CONCLUSIONS:: ALK kinase splicing isoforms were present in NSCLC and even if translated seemed to be nonfunctional variants of ALK.

Original languageEnglish
Pages (from-to)248-253
Number of pages6
JournalJournal of Thoracic Oncology
Volume9
Issue number2
DOIs
Publication statusPublished - 2014 Feb
Externally publishedYes

Fingerprint

Non-Small Cell Lung Carcinoma
Protein Isoforms
Phosphotransferases
anaplastic lymphoma kinase
Exons
MAP4
Mutation
Phosphotyrosine

Keywords

  • Alternative splicing
  • Anaplastic lymphoma kinase
  • Crizotinib
  • Exon 23
  • Exon 27
  • Exon skipping
  • Kinase inhibitor
  • Lung cancer
  • Non-small-cell lung cancer
  • Tyrosine kinase

ASJC Scopus subject areas

  • Oncology
  • Pulmonary and Respiratory Medicine

Cite this

De Figueiredo-Pontes, L. L., Wong, D. W. S., Tin, V. P. C., Chung, L. P., Yasuda, H., Yamaguchi, N., ... Costa, D. B. (2014). Identification and Characterization of ALK Kinase Splicing Isoforms in Non-Small-Cell Lung Cancer. Journal of Thoracic Oncology, 9(2), 248-253. https://doi.org/10.1097/JTO.0000000000000050

Identification and Characterization of ALK Kinase Splicing Isoforms in Non-Small-Cell Lung Cancer. / De Figueiredo-Pontes, Lorena Lobo; Wong, Daisy Wing Sze; Tin, Vicky Pui Chi; Chung, Lap Ping; Yasuda, Hiroyuki; Yamaguchi, Norihiro; Nakayama, Sohei; Jänne, Pasi Antero; Wong, Maria Pik; Kobayashi, Susumu Soeda; Costa, Daniel Botelho.

In: Journal of Thoracic Oncology, Vol. 9, No. 2, 02.2014, p. 248-253.

Research output: Contribution to journalArticle

De Figueiredo-Pontes, LL, Wong, DWS, Tin, VPC, Chung, LP, Yasuda, H, Yamaguchi, N, Nakayama, S, Jänne, PA, Wong, MP, Kobayashi, SS & Costa, DB 2014, 'Identification and Characterization of ALK Kinase Splicing Isoforms in Non-Small-Cell Lung Cancer', Journal of Thoracic Oncology, vol. 9, no. 2, pp. 248-253. https://doi.org/10.1097/JTO.0000000000000050
De Figueiredo-Pontes, Lorena Lobo ; Wong, Daisy Wing Sze ; Tin, Vicky Pui Chi ; Chung, Lap Ping ; Yasuda, Hiroyuki ; Yamaguchi, Norihiro ; Nakayama, Sohei ; Jänne, Pasi Antero ; Wong, Maria Pik ; Kobayashi, Susumu Soeda ; Costa, Daniel Botelho. / Identification and Characterization of ALK Kinase Splicing Isoforms in Non-Small-Cell Lung Cancer. In: Journal of Thoracic Oncology. 2014 ; Vol. 9, No. 2. pp. 248-253.
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abstract = "INTRODUCTION:: Anaplastic lymphoma kinase (ALK) rearrangements are present in an important subset of non-small-cell lung cancer (NSCLC) and predict for response to the tyrosine kinase inhibitor crizotinib. In this study, we evaluated the yet unknown frequency and functional role of ALK splicing isoforms in NSCLC. METHODS:: We analyzed 270 cases of NSCLC for ALK kinase domain splicing aberrations and in addition generated constructs with full-length echinoderm microtubule-associated protein-like 4 (EML4)-ALK (E13;A20) and a splicing isoform. RESULTS:: Splicing isoforms of the kinase domain of ALK - including complete skipping of exon 23 (ALKdel23, ALK p.I1171fs*42) and exon 27 (ALKdel27, ALK p.T1312fs*0) - were identified in 11.1{\%} (30 of 270 cases) of NSCLC, and these changes coexisted with ALK rearrangements, KRAS mutations, and EGFR mutations. ALK splicing isoforms were observed with full-length EML4-ALK in crizotinib-naive and treated NSCLCs. ALK T1312fs*0 was unable to render cells solely dependent on ALK signaling. Unlike EML4-ALK and EML4-ALK p.L1196M, EML4-ALK T1312fs*0 did not autophosphorylate ALK or other phosphotyrosine sites. Coexpression of equal amounts of EML4-ALK T1312fs*0 and EML4-ALK did not result in resistance to crizotinib, whereas coexpression of EML4-ALK L1196M with EML4-ALK resulted in resistance to inhibition of ALK by crizotinib. CONCLUSIONS:: ALK kinase splicing isoforms were present in NSCLC and even if translated seemed to be nonfunctional variants of ALK.",
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AU - De Figueiredo-Pontes, Lorena Lobo

AU - Wong, Daisy Wing Sze

AU - Tin, Vicky Pui Chi

AU - Chung, Lap Ping

AU - Yasuda, Hiroyuki

AU - Yamaguchi, Norihiro

AU - Nakayama, Sohei

AU - Jänne, Pasi Antero

AU - Wong, Maria Pik

AU - Kobayashi, Susumu Soeda

AU - Costa, Daniel Botelho

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N2 - INTRODUCTION:: Anaplastic lymphoma kinase (ALK) rearrangements are present in an important subset of non-small-cell lung cancer (NSCLC) and predict for response to the tyrosine kinase inhibitor crizotinib. In this study, we evaluated the yet unknown frequency and functional role of ALK splicing isoforms in NSCLC. METHODS:: We analyzed 270 cases of NSCLC for ALK kinase domain splicing aberrations and in addition generated constructs with full-length echinoderm microtubule-associated protein-like 4 (EML4)-ALK (E13;A20) and a splicing isoform. RESULTS:: Splicing isoforms of the kinase domain of ALK - including complete skipping of exon 23 (ALKdel23, ALK p.I1171fs*42) and exon 27 (ALKdel27, ALK p.T1312fs*0) - were identified in 11.1% (30 of 270 cases) of NSCLC, and these changes coexisted with ALK rearrangements, KRAS mutations, and EGFR mutations. ALK splicing isoforms were observed with full-length EML4-ALK in crizotinib-naive and treated NSCLCs. ALK T1312fs*0 was unable to render cells solely dependent on ALK signaling. Unlike EML4-ALK and EML4-ALK p.L1196M, EML4-ALK T1312fs*0 did not autophosphorylate ALK or other phosphotyrosine sites. Coexpression of equal amounts of EML4-ALK T1312fs*0 and EML4-ALK did not result in resistance to crizotinib, whereas coexpression of EML4-ALK L1196M with EML4-ALK resulted in resistance to inhibition of ALK by crizotinib. CONCLUSIONS:: ALK kinase splicing isoforms were present in NSCLC and even if translated seemed to be nonfunctional variants of ALK.

AB - INTRODUCTION:: Anaplastic lymphoma kinase (ALK) rearrangements are present in an important subset of non-small-cell lung cancer (NSCLC) and predict for response to the tyrosine kinase inhibitor crizotinib. In this study, we evaluated the yet unknown frequency and functional role of ALK splicing isoforms in NSCLC. METHODS:: We analyzed 270 cases of NSCLC for ALK kinase domain splicing aberrations and in addition generated constructs with full-length echinoderm microtubule-associated protein-like 4 (EML4)-ALK (E13;A20) and a splicing isoform. RESULTS:: Splicing isoforms of the kinase domain of ALK - including complete skipping of exon 23 (ALKdel23, ALK p.I1171fs*42) and exon 27 (ALKdel27, ALK p.T1312fs*0) - were identified in 11.1% (30 of 270 cases) of NSCLC, and these changes coexisted with ALK rearrangements, KRAS mutations, and EGFR mutations. ALK splicing isoforms were observed with full-length EML4-ALK in crizotinib-naive and treated NSCLCs. ALK T1312fs*0 was unable to render cells solely dependent on ALK signaling. Unlike EML4-ALK and EML4-ALK p.L1196M, EML4-ALK T1312fs*0 did not autophosphorylate ALK or other phosphotyrosine sites. Coexpression of equal amounts of EML4-ALK T1312fs*0 and EML4-ALK did not result in resistance to crizotinib, whereas coexpression of EML4-ALK L1196M with EML4-ALK resulted in resistance to inhibition of ALK by crizotinib. CONCLUSIONS:: ALK kinase splicing isoforms were present in NSCLC and even if translated seemed to be nonfunctional variants of ALK.

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KW - Exon 27

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KW - Tyrosine kinase

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