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Updated: May 17, 2026
Zebrafish Model of Neuroblastoma Metastasis
Published on: March 14, 2021
Molecular evidence for the bi-clonal origin of neuroendocrine tumor derived metastases
Beate Rinner1, Birgit Gallè, Slave Trajanoski
1Center for Medical Research, Medical University of Graz, Stiftingtalstraße 24, Graz, 8010, Austria.
Background:
Reports on common mutations in neuroendocrine tumors (NET) are rare and clonality of NET metastases has not been investigated in this tumor entity yet. We selected one NET and the corresponding lymph node and liver metastases as well as the derivative cell lines to screen for somatic mutations in the primary NET and to track the fate of genetic changes during metastasis and in vitro progression.
Results:
Applying microarray based sequence capture resequencing including 4,935 Exons from of 203 cancer-associated genes and high-resolution copy number and genotype analysis identified multiple somatic mutations in the primary NET, affecting BRCA2, CTNNB1, ERCC5, HNF1A, KIT, MLL, RB1, ROS1, SMAD4, and TP53. All mutations were confirmed in the patients' lymph node and liver metastasis tissue as well as early cell line passages. In contrast to the tumor derived cell line, higher passages of the metastases derived cell lines lacked somatic mutations and chromosomal alterations, while expression of the classical NET marker serotonin was maintained.
Conclusion:
Our study reveals that both metastases have evolved from the same pair of genetically differing NET cell clones. In both metastases, the in vivo dominating "mutant" tumor cell clone has undergone negative selection in vitro being replaced by the "non-mutant" tumor cell population. This is the first report of a bi-clonal origin of NET derived metastases, indicating selective advantage of interclonal cooperation during metastasis. In addition, this study underscores the importance to monitor cell line integrity using high-resolution genome analysis tools.
Insights
Neuroendocrine tumors (NET) metastases show a bi-clonal origin, with differing cell clones cooperating during metastasis. In vitro progression leads to negative selection of mutant clones in these NETs.
Area of Science:
- Oncology
- Genetics
- Cancer Biology
Background:
- Limited data exists on common mutations in neuroendocrine tumors (NETs).
- Clonality of NET metastases remains largely uninvestigated.
- This study examines genetic alterations in primary NETs and their metastases.
Purpose of the Study:
- To identify somatic mutations in primary NETs.
- To track genetic changes during metastasis and in vitro progression.
- To investigate the clonality and origin of NET metastases.
Main Methods:
- Microarray-based sequence capture resequencing of 203 cancer-associated genes.
- High-resolution copy number and genotype analysis.
- Analysis of primary NET, metastases, and derived cell lines.
Main Results:
- Multiple somatic mutations identified in primary NETs (BRCA2, CTNNB1, ERCC5, HNF1A, KIT, MLL, RB1, ROS1, SMAD4, TP53).
- All mutations were confirmed in metastases and early cell line passages.
- Higher cell line passages showed loss of somatic mutations and chromosomal alterations.
Conclusions:
- NET metastases originate from a bi-clonal population of genetically distinct NET cell clones.
- In vitro culture leads to negative selection of the 'mutant' clone, favoring the 'non-mutant' clone.
- Highlights the importance of monitoring cell line integrity using genomic analysis.
