Telomere elongation through hTERT immortalization leads to chromosome repositioning in control cells and genomic instability in Hutchinson-Gilford progeria syndrome fibroblasts, expressing a novel SUN1 isoform

Mehmet U. Bikkul, Richard G.A. Faragher, Gemma Worthington, Peter Meinke, Alastair R.W. Kerr, Aakila Sammy, Kumars Riyahi, Daniel Horton, Eric C. Schirmer, Michael Hubank, Ian R. Kill, Rhona M. Anderson, Predrag Slijepcevic, Evgeny Makarov, Joanna M. Bridger

Research output: Contribution to journalArticle

Abstract

Immortalizing primary cells with human telomerase reverse transcriptase (hTERT) has been common practice to enable primary cells to be of extended use in the laboratory because they avoid replicative senescence. Studying exogenously expressed hTERT in cells also affords scientists models of early carcinogenesis and telomere behavior. Control and the premature ageing disease—Hutchinson-Gilford progeria syndrome (HGPS) primary dermal fibroblasts, with and without the classical G608G mutation have been immortalized with exogenous hTERT. However, hTERT immortalization surprisingly elicits genome reorganization not only in disease cells but also in the normal control cells, such that whole chromosome territories normally located at the nuclear periphery in proliferating fibroblasts become mislocalized in the nuclear interior. This includes chromosome 18 in the control fibroblasts and both chromosomes 18 and X in HGPS cells, which physically express an isoform of the LINC complex protein SUN1 that has previously only been theoretical. Additionally, this HGPS cell line has also become genomically unstable and has a tetraploid karyotype, which could be due to the novel SUN1 isoform. Long-term treatment with the hTERT inhibitor BIBR1532 enabled the reduction of telomere length in the immortalized cells and resulted that these mislocalized internal chromosomes to be located at the nuclear periphery, as assessed in actively proliferating cells. Taken together, these findings reveal that elongated telomeres lead to dramatic chromosome mislocalization, which can be restored with a drug treatment that results in telomere reshortening and that a novel SUN1 isoform combined with elongated telomeres leads to genomic instability. Thus, care should be taken when interpreting data from genomic studies in hTERT-immortalized cell lines.

Original languageEnglish
Pages (from-to)341-356
JournalGenes Chromosomes and Cancer
Volume58
Issue number6
DOIs
Publication statusPublished - 26 Nov 2018

Bibliographical note

© 2018 The Authors. Genes, Chromosomes & Cancer published by Wiley Periodicals, Inc. This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

Keywords

  • BIBR1532
  • chromosome territories
  • genomic instability
  • hTERT
  • Hutchinson-Gilford progeria syndrome
  • M-FISH
  • Q-FISH
  • SUN1
  • SUN1 isoform 9
  • telomeres

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    Bikkul, M. U., Faragher, R. G. A., Worthington, G., Meinke, P., Kerr, A. R. W., Sammy, A., Riyahi, K., Horton, D., Schirmer, E. C., Hubank, M., Kill, I. R., Anderson, R. M., Slijepcevic, P., Makarov, E., & Bridger, J. M. (2018). Telomere elongation through hTERT immortalization leads to chromosome repositioning in control cells and genomic instability in Hutchinson-Gilford progeria syndrome fibroblasts, expressing a novel SUN1 isoform. Genes Chromosomes and Cancer, 58(6), 341-356. https://doi.org/10.1002/gcc.22711