Telomerase variant A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal carcinomas

Yuwei Zhang, Rodrigo Calado, Mahadev Rao, Julie A. Hong, Alan K. Meeker, Bogdan Dumitriu, Scott Atay, Peter J. McCormick, Susan H. Garfield, Danny Wangsa, Hesed M. Padilla-Nash, Sandra Burkett, Mary Zhang, Tricia F. Kunst, Nathan R. Peterson, Sichuan Xi, Suzanne Inchauste, Nasser K. Altorki, Alan G. Casson, David G. Beer & 4 others Curtis C. Harris, Thomas Ried, Neal S. Young, David S. Schrump

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Abstract

Background: Although implicated in the pathogenesis of several chronic inflammatory disorders and hematologic malignancies, telomerase mutations have not been thoroughly characterized in human cancers. The present study was performed to examine the frequency and potential clinical relevance of telomerase mutations in esophageal carcinomas. Methods: Sequencing techniques were used to evaluate mutational status of telomerase reverse transcriptase (TERT) and telomerase RNA component (TERC) in neoplastic and adjacent normal mucosa from 143 esophageal cancer (EsC) patients. MTS, flow cytometry, time lapse microscopy, and murine xenograft techniques were used to assess proliferation, apoptosis, chemotaxis, and tumorigenicity of EsC cells expressing either wtTERT or TERT variants. Immunoprecipitation, immunoblot, immunofluorescence, promoter-reporter and qRT-PCR techniques were used to evaluate interactions of TERT and several TERT variants with BRG-1 and β-catenin, and to assess expression of cytoskeletal proteins, and cell signaling. Fluorescence in-situ hybridization and spectral karyotyping techniques were used to examine telomere length and chromosomal stability. Results: Sequencing analysis revealed one deletion involving TERC (TERC del 341-360), and two non-synonymous TERT variants [A279T (2 homozygous, 9 heterozygous); A1062T (4 heterozygous)]. The minor allele frequency of the A279T variant was five-fold higher in EsC patients compared to healthy blood donors (p<0.01). Relative to wtTERT, A279T decreased telomere length, destabilized TERT-BRG-1-β-catenin complex, markedly depleted β-catenin, and down-regulated canonical Wnt signaling in cancer cells; these phenomena coincided with decreased proliferation, depletion of additional cytoskeletal proteins, impaired chemotaxis, increased chemosensitivity, and significantly decreased tumorigenicity of EsC cells. A279T expression significantly increased chromosomal aberrations in mouse embryonic fibroblasts (MEFs) following Zeocin™ exposure, as well as Li Fraumeni fibroblasts in the absence of pharmacologically-induced DNA damage. Conclusions: A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal cancer cells. These findings warrant further analysis of A279T expression in esophageal cancers and premalignant esophageal lesions.

Original languageEnglish
Article numbere101010
JournalPLoS One
Volume9
Issue number7
DOIs
Publication statusPublished - 01-07-2014

Fingerprint

telomerase
Telomerase
telomeres
Telomere
carcinoma
Esophageal Neoplasms
Carcinoma
esophageal neoplasms
RNA-directed DNA polymerase
Catenins
Cytoskeletal Proteins
Cells
Chemotaxis
Fibroblasts
cytoskeletal proteins
chemotaxis
RNA
Spectral Karyotyping
Cell signaling
fibroblasts

All Science Journal Classification (ASJC) codes

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Zhang, Yuwei ; Calado, Rodrigo ; Rao, Mahadev ; Hong, Julie A. ; Meeker, Alan K. ; Dumitriu, Bogdan ; Atay, Scott ; McCormick, Peter J. ; Garfield, Susan H. ; Wangsa, Danny ; Padilla-Nash, Hesed M. ; Burkett, Sandra ; Zhang, Mary ; Kunst, Tricia F. ; Peterson, Nathan R. ; Xi, Sichuan ; Inchauste, Suzanne ; Altorki, Nasser K. ; Casson, Alan G. ; Beer, David G. ; Harris, Curtis C. ; Ried, Thomas ; Young, Neal S. ; Schrump, David S. / Telomerase variant A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal carcinomas. In: PLoS One. 2014 ; Vol. 9, No. 7.
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title = "Telomerase variant A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal carcinomas",
abstract = "Background: Although implicated in the pathogenesis of several chronic inflammatory disorders and hematologic malignancies, telomerase mutations have not been thoroughly characterized in human cancers. The present study was performed to examine the frequency and potential clinical relevance of telomerase mutations in esophageal carcinomas. Methods: Sequencing techniques were used to evaluate mutational status of telomerase reverse transcriptase (TERT) and telomerase RNA component (TERC) in neoplastic and adjacent normal mucosa from 143 esophageal cancer (EsC) patients. MTS, flow cytometry, time lapse microscopy, and murine xenograft techniques were used to assess proliferation, apoptosis, chemotaxis, and tumorigenicity of EsC cells expressing either wtTERT or TERT variants. Immunoprecipitation, immunoblot, immunofluorescence, promoter-reporter and qRT-PCR techniques were used to evaluate interactions of TERT and several TERT variants with BRG-1 and β-catenin, and to assess expression of cytoskeletal proteins, and cell signaling. Fluorescence in-situ hybridization and spectral karyotyping techniques were used to examine telomere length and chromosomal stability. Results: Sequencing analysis revealed one deletion involving TERC (TERC del 341-360), and two non-synonymous TERT variants [A279T (2 homozygous, 9 heterozygous); A1062T (4 heterozygous)]. The minor allele frequency of the A279T variant was five-fold higher in EsC patients compared to healthy blood donors (p<0.01). Relative to wtTERT, A279T decreased telomere length, destabilized TERT-BRG-1-β-catenin complex, markedly depleted β-catenin, and down-regulated canonical Wnt signaling in cancer cells; these phenomena coincided with decreased proliferation, depletion of additional cytoskeletal proteins, impaired chemotaxis, increased chemosensitivity, and significantly decreased tumorigenicity of EsC cells. A279T expression significantly increased chromosomal aberrations in mouse embryonic fibroblasts (MEFs) following Zeocin™ exposure, as well as Li Fraumeni fibroblasts in the absence of pharmacologically-induced DNA damage. Conclusions: A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal cancer cells. These findings warrant further analysis of A279T expression in esophageal cancers and premalignant esophageal lesions.",
author = "Yuwei Zhang and Rodrigo Calado and Mahadev Rao and Hong, {Julie A.} and Meeker, {Alan K.} and Bogdan Dumitriu and Scott Atay and McCormick, {Peter J.} and Garfield, {Susan H.} and Danny Wangsa and Padilla-Nash, {Hesed M.} and Sandra Burkett and Mary Zhang and Kunst, {Tricia F.} and Peterson, {Nathan R.} and Sichuan Xi and Suzanne Inchauste and Altorki, {Nasser K.} and Casson, {Alan G.} and Beer, {David G.} and Harris, {Curtis C.} and Thomas Ried and Young, {Neal S.} and Schrump, {David S.}",
year = "2014",
month = "7",
day = "1",
doi = "10.1371/journal.pone.0101010",
language = "English",
volume = "9",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "7",

}

Zhang, Y, Calado, R, Rao, M, Hong, JA, Meeker, AK, Dumitriu, B, Atay, S, McCormick, PJ, Garfield, SH, Wangsa, D, Padilla-Nash, HM, Burkett, S, Zhang, M, Kunst, TF, Peterson, NR, Xi, S, Inchauste, S, Altorki, NK, Casson, AG, Beer, DG, Harris, CC, Ried, T, Young, NS & Schrump, DS 2014, 'Telomerase variant A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal carcinomas', PLoS One, vol. 9, no. 7, e101010. https://doi.org/10.1371/journal.pone.0101010

Telomerase variant A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal carcinomas. / Zhang, Yuwei; Calado, Rodrigo; Rao, Mahadev; Hong, Julie A.; Meeker, Alan K.; Dumitriu, Bogdan; Atay, Scott; McCormick, Peter J.; Garfield, Susan H.; Wangsa, Danny; Padilla-Nash, Hesed M.; Burkett, Sandra; Zhang, Mary; Kunst, Tricia F.; Peterson, Nathan R.; Xi, Sichuan; Inchauste, Suzanne; Altorki, Nasser K.; Casson, Alan G.; Beer, David G.; Harris, Curtis C.; Ried, Thomas; Young, Neal S.; Schrump, David S.

In: PLoS One, Vol. 9, No. 7, e101010, 01.07.2014.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Telomerase variant A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal carcinomas

AU - Zhang, Yuwei

AU - Calado, Rodrigo

AU - Rao, Mahadev

AU - Hong, Julie A.

AU - Meeker, Alan K.

AU - Dumitriu, Bogdan

AU - Atay, Scott

AU - McCormick, Peter J.

AU - Garfield, Susan H.

AU - Wangsa, Danny

AU - Padilla-Nash, Hesed M.

AU - Burkett, Sandra

AU - Zhang, Mary

AU - Kunst, Tricia F.

AU - Peterson, Nathan R.

AU - Xi, Sichuan

AU - Inchauste, Suzanne

AU - Altorki, Nasser K.

AU - Casson, Alan G.

AU - Beer, David G.

AU - Harris, Curtis C.

AU - Ried, Thomas

AU - Young, Neal S.

AU - Schrump, David S.

PY - 2014/7/1

Y1 - 2014/7/1

N2 - Background: Although implicated in the pathogenesis of several chronic inflammatory disorders and hematologic malignancies, telomerase mutations have not been thoroughly characterized in human cancers. The present study was performed to examine the frequency and potential clinical relevance of telomerase mutations in esophageal carcinomas. Methods: Sequencing techniques were used to evaluate mutational status of telomerase reverse transcriptase (TERT) and telomerase RNA component (TERC) in neoplastic and adjacent normal mucosa from 143 esophageal cancer (EsC) patients. MTS, flow cytometry, time lapse microscopy, and murine xenograft techniques were used to assess proliferation, apoptosis, chemotaxis, and tumorigenicity of EsC cells expressing either wtTERT or TERT variants. Immunoprecipitation, immunoblot, immunofluorescence, promoter-reporter and qRT-PCR techniques were used to evaluate interactions of TERT and several TERT variants with BRG-1 and β-catenin, and to assess expression of cytoskeletal proteins, and cell signaling. Fluorescence in-situ hybridization and spectral karyotyping techniques were used to examine telomere length and chromosomal stability. Results: Sequencing analysis revealed one deletion involving TERC (TERC del 341-360), and two non-synonymous TERT variants [A279T (2 homozygous, 9 heterozygous); A1062T (4 heterozygous)]. The minor allele frequency of the A279T variant was five-fold higher in EsC patients compared to healthy blood donors (p<0.01). Relative to wtTERT, A279T decreased telomere length, destabilized TERT-BRG-1-β-catenin complex, markedly depleted β-catenin, and down-regulated canonical Wnt signaling in cancer cells; these phenomena coincided with decreased proliferation, depletion of additional cytoskeletal proteins, impaired chemotaxis, increased chemosensitivity, and significantly decreased tumorigenicity of EsC cells. A279T expression significantly increased chromosomal aberrations in mouse embryonic fibroblasts (MEFs) following Zeocin™ exposure, as well as Li Fraumeni fibroblasts in the absence of pharmacologically-induced DNA damage. Conclusions: A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal cancer cells. These findings warrant further analysis of A279T expression in esophageal cancers and premalignant esophageal lesions.

AB - Background: Although implicated in the pathogenesis of several chronic inflammatory disorders and hematologic malignancies, telomerase mutations have not been thoroughly characterized in human cancers. The present study was performed to examine the frequency and potential clinical relevance of telomerase mutations in esophageal carcinomas. Methods: Sequencing techniques were used to evaluate mutational status of telomerase reverse transcriptase (TERT) and telomerase RNA component (TERC) in neoplastic and adjacent normal mucosa from 143 esophageal cancer (EsC) patients. MTS, flow cytometry, time lapse microscopy, and murine xenograft techniques were used to assess proliferation, apoptosis, chemotaxis, and tumorigenicity of EsC cells expressing either wtTERT or TERT variants. Immunoprecipitation, immunoblot, immunofluorescence, promoter-reporter and qRT-PCR techniques were used to evaluate interactions of TERT and several TERT variants with BRG-1 and β-catenin, and to assess expression of cytoskeletal proteins, and cell signaling. Fluorescence in-situ hybridization and spectral karyotyping techniques were used to examine telomere length and chromosomal stability. Results: Sequencing analysis revealed one deletion involving TERC (TERC del 341-360), and two non-synonymous TERT variants [A279T (2 homozygous, 9 heterozygous); A1062T (4 heterozygous)]. The minor allele frequency of the A279T variant was five-fold higher in EsC patients compared to healthy blood donors (p<0.01). Relative to wtTERT, A279T decreased telomere length, destabilized TERT-BRG-1-β-catenin complex, markedly depleted β-catenin, and down-regulated canonical Wnt signaling in cancer cells; these phenomena coincided with decreased proliferation, depletion of additional cytoskeletal proteins, impaired chemotaxis, increased chemosensitivity, and significantly decreased tumorigenicity of EsC cells. A279T expression significantly increased chromosomal aberrations in mouse embryonic fibroblasts (MEFs) following Zeocin™ exposure, as well as Li Fraumeni fibroblasts in the absence of pharmacologically-induced DNA damage. Conclusions: A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal cancer cells. These findings warrant further analysis of A279T expression in esophageal cancers and premalignant esophageal lesions.

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