Reliable assessment of telomere maintenance mechanisms in neuroblastoma.
Alternative lengthening of telomeres
Neuroblastoma
Telomerase
Telomere maintenance
Journal
Cell & bioscience
ISSN: 2045-3701
Titre abrégé: Cell Biosci
Pays: England
ID NLM: 101561195
Informations de publication
Date de publication:
24 Sep 2022
24 Sep 2022
Historique:
received:
29
05
2022
accepted:
03
09
2022
entrez:
24
9
2022
pubmed:
25
9
2022
medline:
25
9
2022
Statut:
epublish
Résumé
Telomere maintenance mechanisms (TMM) are a hallmark of high-risk neuroblastoma, and are conferred by activation of telomerase or alternative lengthening of telomeres (ALT). However, detection of TMM is not yet part of the clinical routine, and consensus on TMM detection, especially on ALT assessment, remains to be achieved. Whole genome sequencing (WGS) data of 68 primary neuroblastoma samples were analyzed. Telomere length was calculated from WGS data or by telomere restriction fragment analysis (n = 39). ALT was assessed by C-circle assay (CCA, n = 67) and detection of ALT-associated PML nuclear bodies (APB) by combined fluorescence in situ hybridization and immunofluorescence staining (n = 68). RNA sequencing was performed (n = 64) to determine expression of TERT and telomeric long non-coding RNA (TERRA). Telomerase activity was examined by telomerase repeat amplification protocol (TRAP, n = 15). Tumors were considered as telomerase-positive if they harbored a TERT rearrangement, MYCN amplification or high TERT expression (45.6%, 31/68), and ALT-positive if they were positive for APB and CCA (19.1%, 13/68). If all these markers were absent, tumors were considered TMM-negative (25.0%, 17/68). According to these criteria, the majority of samples were classified unambiguously (89.7%, 61/68). Assessment of additional ALT-associated parameters clarified the TMM status of the remaining seven cases with high likelihood: ALT-positive tumors had higher TERRA expression, longer telomeres, more telomere insertions, a characteristic pattern of telomere variant repeats, and were associated with ATRX mutations. We here propose a workflow to reliably detect TMM in neuroblastoma. We show that unambiguous classification is feasible following a stepwise approach that determines both, activation of telomerase and ALT. The workflow proposed in this study can be used in clinical routine and provides a framework to systematically and reliably determine telomere maintenance mechanisms for risk stratification and treatment allocation of neuroblastoma patients.
Sections du résumé
BACKGROUND
BACKGROUND
Telomere maintenance mechanisms (TMM) are a hallmark of high-risk neuroblastoma, and are conferred by activation of telomerase or alternative lengthening of telomeres (ALT). However, detection of TMM is not yet part of the clinical routine, and consensus on TMM detection, especially on ALT assessment, remains to be achieved.
METHODS
METHODS
Whole genome sequencing (WGS) data of 68 primary neuroblastoma samples were analyzed. Telomere length was calculated from WGS data or by telomere restriction fragment analysis (n = 39). ALT was assessed by C-circle assay (CCA, n = 67) and detection of ALT-associated PML nuclear bodies (APB) by combined fluorescence in situ hybridization and immunofluorescence staining (n = 68). RNA sequencing was performed (n = 64) to determine expression of TERT and telomeric long non-coding RNA (TERRA). Telomerase activity was examined by telomerase repeat amplification protocol (TRAP, n = 15).
RESULTS
RESULTS
Tumors were considered as telomerase-positive if they harbored a TERT rearrangement, MYCN amplification or high TERT expression (45.6%, 31/68), and ALT-positive if they were positive for APB and CCA (19.1%, 13/68). If all these markers were absent, tumors were considered TMM-negative (25.0%, 17/68). According to these criteria, the majority of samples were classified unambiguously (89.7%, 61/68). Assessment of additional ALT-associated parameters clarified the TMM status of the remaining seven cases with high likelihood: ALT-positive tumors had higher TERRA expression, longer telomeres, more telomere insertions, a characteristic pattern of telomere variant repeats, and were associated with ATRX mutations.
CONCLUSIONS
CONCLUSIONS
We here propose a workflow to reliably detect TMM in neuroblastoma. We show that unambiguous classification is feasible following a stepwise approach that determines both, activation of telomerase and ALT. The workflow proposed in this study can be used in clinical routine and provides a framework to systematically and reliably determine telomere maintenance mechanisms for risk stratification and treatment allocation of neuroblastoma patients.
Identifiants
pubmed: 36153564
doi: 10.1186/s13578-022-00896-2
pii: 10.1186/s13578-022-00896-2
pmc: PMC9508734
doi:
Types de publication
Journal Article
Langues
eng
Pagination
160Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : FI 1926/2-1
Organisme : Deutsche Forschungsgemeinschaft
ID : as part of the SFB1399
Organisme : Deutsche Forschungsgemeinschaft
ID : BA 6984/1-1
Organisme : Else Kröner-Fresenius-Stiftung
ID : 2016-Kolleg-19
Organisme : Bundesministerium für Bildung und Forschung
ID : 01ZX1303
Organisme : Bundesministerium für Bildung und Forschung
ID : 01ZX1603
Organisme : Bundesministerium für Bildung und Forschung
ID : 01ZX1307
Organisme : Bundesministerium für Bildung und Forschung
ID : 01ZX1607
Organisme : Universität zu Köln
ID : Köln Fortune Program/Faculty of Medicine
Informations de copyright
© 2022. The Author(s).
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