Ragnhild Adelheid Lothe

• Yaqub, Sheraz; Valberg, Morten; Bjørnbeth, Bjørn Atle; Angelsen, Jon-Helge; Brudvik, Kristoffer Watten & Eilertsen, Ina [Show all 43 contributors for this article] (2026). Aspirin after curative-intent treatment of colorectal cancer liver metastases (ASAC): a multicentre, phase 3, randomised, double-blind, placebo-controlled trial. The Lancet Gastroenterology and Hepatology. ISSN 2468-1253. doi: 10.1016/s2468-1253(26)00260-8. Full text in Research Archive Show summary

  Background Colorectal cancer frequently metastasises to the liver. Recurrence after curative-intent treatment of colorectal cancer liver metastases is common. Aspirin has shown preventive and antitumour effects in colorectal cancer, but its role after treatment of metastatic disease is uncertain. We aimed to establish whether adjuvant aspirin improves disease-free survival after curative-intent treatment of colorectal cancer liver metastases. Methods In this phase 3, randomised, double-blind, placebo-controlled trial conducted at 14 centres in Norway, Sweden, and Denmark, patients who had undergone curative-intent treatment for colorectal cancer liver metastases were randomly assigned (1:1), stratified by study site using a centrally generated computerised sequence with variable block sizes, to aspirin 160 mg per day or matching placebo for up to 3 years or until recurrence, death, or treatment discontinuation. The primary outcome was disease-free survival in the full analysis set, comprising all patients who initiated study treatment. The trial is completed and this is the final analysis. This trial is registered with ClinicalTrials.gov, NCT03326791, and EudraCT, 2014-003601-15. Findings Between Dec 15, 2017, and Jan 27, 2022, 466 patients were randomly assigned, of whom 428 initiated study treatment and were included in the full analysis set (217 assigned aspirin and 211 assigned placebo); 272 (64%) were male, 156 (36%) were female, and the mean age was 62·2 years. 269 disease-free survival events occurred (138 in the aspirin group and 131 in the placebo group). Aspirin did not improve disease-free survival (hazard ratio [HR] for recurrence or death 1·08 [95·44% CI 0·85–1·38]; p=0·58). Median disease-free survival was 1·13 years (95·44% CI 0·92–1·44) with aspirin and 1·40 years (1·11–1·69) with placebo. Overall survival at 36 months was 76% (95% CI 69·4–82·7) with aspirin and 85% (79·5–90·8) with placebo (HR for death 1·64 [95% CI 1·01–2·65]; p=0·045). Time to recurrence did not differ between groups (adjusted cause-specific HR 1·06 [95% CI 0·83–1·35]; p=0·64). 24 serious adverse events occurred in 17 (8%) of 217 patients in the aspirin group and five serious adverse events occurred in four (2%) of 211 patients in the placebo group. Clinically relevant adverse events and serious adverse events that occurred more commonly in the aspirin group than in the placebo group were myocardial infarction (two vs 0), epistaxis (four vs 0), duodenal ulcer (two vs 0), and cerebral haemorrhage (two vs 0). There were no treatment-related deaths. Interpretation Adjuvant aspirin did not improve disease-free survival and was associated with lower overall survival and more serious adverse events. These findings do not support routine use of aspirin after curative-intent treatment of colorectal cancer liver metastases.

• Panagopoulos, Ioannis; Stranger, Ane; Sveen, Anita; Jareid, Mie Linnea; Kidd, Susanne Gundersen & Bischof, Katharina [Show all 12 contributors for this article] (2026). Fusion gene heterogeneity and kinase enrichment in high-grade serous carcinomas. Neoplasia. ISSN 1522-8002. 79, p. 101332–101332. doi: 10.1016/j.neo.2026.101332. Full text in Research Archive
• Nunes, Luis; Nesbakken, Arild; Lothe, Ragnhild Adelheid & Sveen, Anita (2026). Temporal Trends in Cancer Mortality Among People Younger Than 50 Years in Norway. JAMA Network Open. 9(8), p. e2629437–e2629437. doi: 10.1001/jamanetworkopen.2026.29437. Full text in Research Archive
• Sveen, Anita; Stenersen, Jakob Mørkved; Asenso, Theophilus Quachie; Klokkerud, Solveig Knoph; Kranjec, Christian & Eilertsen, Ina [Show all 11 contributors for this article] (2026). Exceptional treatment responses and molecular markers among 103 colorectal cancer cell lines. Molecular Biomedicine. 7. doi: 10.1186/s43556-026-00551-z. Full text in Research Archive Show summary

  Patients with colorectal cancer have few molecularly guided treatment options. Cancers with exceptional treatment response are valuable models for discovery of molecular response mechanisms and might reveal new predictive biomarkers to guide treatment selection. However, the rarity of events is a clinical challenge. We performed integrated high-throughput drug sensitivity testing (n = 620 drugs) and molecular analyses of 103 colorectal cancer cell lines to build an in vitro foundation for exceptional treatment responses and their molecular markers. Exceptional responses were scored by a two-way outlier detection approach and were identified in 67 unique cell line-drug pairs (0.13% of all pairs tested). This involved 35 cell lines (34.0%) and 49 drugs (7.9%) representing standard, experimental, and non-oncology drugs of diverse classes. Plausible response mechanisms on the gene, protein, and/or pathway expression levels were identified at baseline in 89.6% of the 67 pairs. Experimental validation confirmed treatment-induced marker suppression in seven of eight selected pairs, including target engagement of kinase inhibitors and downregulation of markers of extended response mechanisms. Recurrent exceptional sensitivity to the same drug in different cell lines involved functionally convergent molecular mechanisms, but rarely precisely the same marker. Most exceptional sensitivities (77.8%) could be predicted with multivariable transcriptomic models. In conclusion, this study provides in vitro support for biomarker-guided prescreening to potentially obtain exceptional treatment response to diverse drugs and therapeutic targets in colorectal cancer. Most exceptional responses have a clear molecular underpinning. The study also provides a large and openly available pharmaco-transcriptomics resource for colorectal cancer cell lines.

• Kryeziu, Kushtrim; Klokkerud, Solveig Knoph; Totland, Max Zachrisson; Moosavi, Hossein; Reims, Henrik Mikael & Bergsland, Christian Holst [Show all 17 contributors for this article] (2026). Patient-derived organoids from metastatic colorectal cancer mirror tumor heterogeneity and predict patient survival and drug sensitivity. Cell Reports Medicine. 7(6). doi: 10.1016/j.xcrm.2026.102840. Full text in Research Archive Show summary

  This study reports the pharmacogenomic analysis of tumor heterogeneity in a living biobank of 213 organoids derived from liver metastases of 102 patients with metastatic colorectal cancer. Successful organoid establishment reflects poorer chemosensitivity and patient survival. Molecular fidelity is confirmed in tumor-organoid pairs, and multi-modal phenotypes are defined based on organoid morphology. Cystic morphology associates with intestinal stem cell markers and greater drug sensitivity, while solid morphology is linked to markers of plasticity and aggressiveness. The potential to identify treatments less affected by tumor heterogeneity is supported by multi-lesion analyses in 65 patients. Clinical translation is illustrated in two prospective cases of pharmacogenomics-guided treatment, including successful chemotherapy rechallenge and targeted therapy resistance in cancers with low and high heterogeneity. All pharmacogenomic data are openly available as a functional oncology resource to support interpretation of ex vivo drug sensitivities into clinical “actionability.”

• Kanetsky, Peter A.; Almstrup, Kristian; Cherlin, Svetlana; Cortessis, Victoria K.; Ferlin, Alberto & Gietema, Jourik A. [Show all 29 contributors for this article] (2026). The Testicular Cancer Consortium (TECAC): Filling Knowledge Gaps in the Genetic Etiology of Testicular Germ Cell Tumors. Andrology. ISSN 2047-2919. doi: 10.1111/andr.70168. Full text in Research Archive
• Kranjec, Christian; Eilertsen, Ina; Nunes, Luis; Moosavi, Hossein; Berg, Kaja Christine Graue & Eknæs, Mette [Show all 12 contributors for this article] (2026). Common gene mutations in 103 authenticated colorectal cancer cell lines. Oncogenesis. 15. doi: 10.1038/s41389-026-00599-0. Full text in Research Archive Show summary

  Colorectal cancer (CRC) cell lines represent the main molecular subtypes of tumors and are valuable models for preclinical investigations. However, cell lines can diverge over time and careful selection of models based on their molecular features is key. We have authenticated 103 commonly used CRC cell lines and present the mutation profiles of 20 CRC-relevant genes sequenced to an average depth of 575 times coverage. The cell lines reflected the distinct mutation patterns of hypermutation phenotypes associated with microsatellite instability and pathogenic POLE mutations. Hypermutated cell lines appeared to have a stronger mutational divergence and more frequent subclonal mutations, while mutations not associated with hypermutation were more frequently homozygous or hemizygous, classified as pathogenic, and subject to stronger selection pressure. Loss of heterozygosity at mutated loci was primarily observed in tumor suppressor genes. Genetic interactions based on co-occurring mutations identified cell lines representative of particularly aggressive subtypes of CRC, including concurrent BRAF p.V600 and truncating APC mutations, as well as APC/TP53/RAS triple mutations with double hits of APC. This study provides a resource to guide the selection of cell lines for functional studies of CRC, and detailed mutation data including classifications of pathogenicity, variant allele frequencies and illustrations of the mutation distribution along the length of encoded proteins are included.

• Sveen, Anita; Bergsland, Christian Holst; Niederdorfer, Barbara; Berstad, Audun Elnæs; Bjerkehagen, Bodil & Boye, Kjetil [Show all 8 contributors for this article] (2026). Pembrolizumab in advanced malignant peripheral nerve sheath tumors: a single-arm phase 2 trial. NPJ precision oncology. 10. doi: 10.1038/s41698-026-01303-6. Full text in Research Archive Show summary

  We report a single-arm phase 2 trial of pembrolizumab in advanced malignant peripheral nerve sheath tumors at Oslo University Hospital. Long-lasting response in 1 of 8 patients was associated with high tumor density of PD-1+CD8+ T cells. Mixed response in another patient was associated with PD-L1 expression, and PD-L1 was mostly not co-expressed on CD68+ macrophages or CD8+ cells. The study was prematurely closed due to a slow accrual rate.

• Ugalde-Morales, Emilio; Wilf, Rona; Pluta, John; Ploner, Alexander; Fan, Mengyao & Damra, Mohammad [Show all 49 contributors for this article] (2025). Identification of genes associated with testicular germ cell tumor susceptibility through a transcriptome-wide association study. American Journal of Human Genetics. ISSN 0002-9297. 112(3), p. 630–643. doi: 10.1016/j.ajhg.2025.01.022. Full text in Research Archive Show summary

  Transcriptome-wide association studies (TWASs) have the potential to identify susceptibility genes associated with testicular germ cell tumors (TGCTs). We conducted a comprehensive TGCT TWAS by integrating genome-wide association study (GWAS) summary data with predicted expression models from normal testis, TGCT tissues, and a cross-tissue panel that encompasses shared regulatory features across 22 normal tissues, including the testis. Gene associations were evaluated while accounting for variant-level effects from GWASs, followed by fine-mapping analyses in regions exhibiting multiple TWAS signals, and finally supplemented by colocalization analysis. Expression and protein patterns of identified TWAS genes were further examined in relevant tissues. Our analysis tested 19,805 gene-disease links, revealing 165 TGCT-associated genes with a false discovery rate of less than 0.01. We prioritized 46 candidate genes by considering GWAS-inflated signals, correlations between neighboring genes, and evidence of colocalization. Among these, 23 genes overlap with 22 GWAS loci, with 7 being associations not previously implicated in TGCT risk. Additionally, 23 genes located within 21 loci are at least 1 Mb away from published GWAS index variants. The 46 prioritized genes display expression levels consistent with expected expression levels in human gonadal cell types and precursor tumor cells and significant enrichment in TGCTs. Additionally, immunohistochemistry revealed protein-level accumulation of two candidate genes, ARID3B and GINM1, in both precursor and tumor cells. These findings enhance our understanding of the genetic predisposition to TGCTs and underscore the importance of further functional investigations into these candidate genes.

• Moosavi, Hossein; Kryeziu, Kushtrim; Eilertsen, Ina; Nunes, Luis; Hektoen, Merete & Niederdorfer, Barbara [Show all 13 contributors for this article] (2025). Molecular prognostic factors for liver transplantation of unresectable metastatic colorectal cancer. British Journal of Surgery. ISSN 0007-1323. 112(4). doi: 10.1093/bjs/znaf072. Full text in Research Archive
• Bogaard, Mari; Strømme, Jonas Meier; Kidd, Susanne Gundersen; Johannessen, Bjarne; Bakken, Anne Cathrine & Lothe, Ragnhild Adelheid [Show all 9 contributors for this article] (2024). GRIN3A: A biomarker associated with a cribriform pattern and poor prognosis in prostate cancer. Neoplasia. ISSN 1522-8002. 55. doi: 10.1016/j.neo.2024.101023. Full text in Research Archive Show summary

  Prostate cancer with a cribriform pattern, including invasive cribriform carcinoma (ICC) and/or intraductal carcinoma (IDC) is associated with a poor prognosis, and the underlying mechanisms are unclear. Therefore, we aimed to identify biomarkers for this feature. Using a radical prostatectomy cohort, we performed within-patient differential expression analyses with RNA sequencing data to compare samples with a cribriform pattern to those with non-cribriform Gleason pattern 4 (NcGP4; n=13). ACSM1, GRIN3A, PCDHB2, and REG4 were identified as differentially expressed, and validation was performed using real-time reverse transcription polymerase chain reaction (n=99; 321 RNA samples) and RNA in situ hybridization on tissue microarrays (n=479; 2047 tissue cores). GRIN3A was significantly higher expressed in cribriform pattern vs. NcGP4, when assessed within the same patient (n=27; p=0.005) and between different patients (n=83; p=0.001). Tissue cores with IDC more often expressed GRIN3A compared to ICC, NcGP4, and benign tissue (52 % vs. ≤ 32 %). When IDC and NcGP4 was compared within the same patient (173 pairs of tissue cores; 54 patients), 38 (22 %) of the tissue microarray core pairs had GRIN3A expression in only IDC, 33 (19 %) had expression in both IDC and NcGP4, 14 (8 %) in only NcGP4 and 88 (51 %) were negative in both entities (p=0.001). GRIN3A was as well associated with biochemical recurrence (log-rank, p=0.002). In conclusion, ectopic GRIN3A expression is an RNA-based biomarker for the presence of cribriform prostate cancer, particularly for IDC.

• Sveen, Anita; Johannessen, Bjarne; Klokkerud, Solveig Margrete Knoph; Kraggerud, Sigrid M.; Meza, Leonardo Zepeda & Bjørnslett, Merete Pauline [Show all 13 contributors for this article] (2024). Evolutionary mode and timing of dissemination of high-grade serous carcinomas. JCI Insight. 9:e170423(3), p. 1–19. doi: 10.1172/jci.insight.170423. Full text in Research Archive Show summary

  Dissemination within the peritoneal cavity is a main determinant of poor patient outcomes from high-grade serous carcinomas (HGSCs). The dissemination process is poorly understood from a cancer evolutionary perspective. We reconstructed the evolutionary trajectories across a median of 5 tumor sites and regions from each of 23 patients based on deep whole-exome sequencing. Polyclonal cancer origin was detected in 1 patient. Ovarian tumors had more complex subclonal architectures than other intraperitoneal tumors in each patient, which indicated that tumors developed earlier in the ovaries. Three common modes of dissemination were identified, including monoclonal or polyclonal dissemination of monophyletic (linear) or polyphyletic (branched) subclones. Mutation profiles of initial or disseminated clones varied greatly among cancers, but recurrent mutations were found in 7 cancer-critical genes, including TP53, BRCA1, BRCA2, and DNMT3A, and in the PI3K/AKT1 pathway. Disseminated clones developed late in the evolutionary trajectory models of most cancers, in particular in cancers with DNA damage repair deficiency. Polyclonal dissemination was predicted to occur predominantly as a single and rapid wave, but chemotherapy exposure was associated with higher genomic diversity of disseminated clones. In conclusion, we described three common evolutionary dissemination modes across HGSCs and proposed factors associated with dissemination diversity.

• Majid, Umair; Bergsland, Christian Holst; Sveen, Anita; Bruun, Jarle; Eilertsen, Ina Andrassy & Bækkevold, Espen Sønderaal [Show all 10 contributors for this article] (2024). The prognostic effect of tumor-associated macrophages in stage I-III colorectal cancer depends on T cell infiltration. Cellular Oncology. ISSN 2211-3428. 47(4), p. 1267–1276. doi: 10.1007/s13402-024-00926-w. Full text in Research Archive Show summary

  Background Tumor-associated macrophages (TAMs) are associated with unfavorable patient prognosis in many cancer types. However, TAMs are a heterogeneous cell population and subsets have been shown to activate tumor-infiltrating T cells and confer a good patient prognosis. Data on the prognostic value of TAMs in colorectal cancer are conflicting. We investigated the prognostic effect of TAMs in relation to tumor-infiltrating T cells in colorectal cancers. Methods The TAM markers CD68 and CD163 were analyzed by multiplex fluorescence immunohistochemistry and digital image analysis on tissue microarrays of 1720 primary colorectal cancers. TAM density in the tumor stroma was scored in relation to T cell density (stromal CD3+ and epithelial CD8+ cells) and analyzed in Cox proportional hazards models of 5-year relapse-free survival. Multivariable survival models included clinicopathological factors, MSI status and BRAFV600E mutation status. Results High TAM density was associated with a favorable 5-year relapse-free survival in a multivariable model of patients with stage I–III tumors (p = 0.004, hazard ratio 0.94, 95% confidence interval 0.90–0.98). However, the prognostic effect was dependent on tumoral T-cell density. High TAM density was associated with a good prognosis in patients who also had high T-cell levels in their tumors, while high TAM density was associated with poorer prognosis in patients with low T-cell levels (pinteraction = 0.0006). This prognostic heterogeneity was found for microsatellite stable tumors separately. Conclusions This study supported a phenotypic heterogeneity of TAMs in colorectal cancer, and showed that combined tumor immunophenotyping of multiple immune cell types improved the prediction of patient prognosis.

• Totland, Max; Knudsen, Lars Mørland; Rasmussen, Nikoline Lander; Omori, Yasufumi; Sørensen, Vigdis & Elster, Vilde Cecilie Wivestad [Show all 17 contributors for this article] (2024). The E3 ubiquitin ligase ITCH negatively regulates intercellular communication via gap junctions by targeting connexin43 for lysosomal degradation. Cellular and Molecular Life Sciences (CMLS). ISSN 1420-682X. 81(1), p. 1–20. doi: 10.1007/s00018-024-05165-8. Full text in Research Archive Show summary

  Intercellular communication via gap junctions has a fundamental role in regulating cell growth and tissue homeostasis, and its dysregulation may be involved in cancer development and radio- and chemotherapy resistance. Connexin43 (Cx43) is the most ubiquitously expressed gap junction channel protein in human tissues. Emerging evidence indicates that dysregulation of the sorting of Cx43 to lysosomes is important in mediating the loss of Cx43-based gap junctions in cancer cells. However, the molecular basis underlying this process is currently poorly understood. Here, we identified the E3 ubiquitin ligase ITCH as a novel regulator of intercellular communication via gap junctions. We demonstrate that ITCH promotes loss of gap junctions in cervical cancer cells, which is associated with increased degradation of Cx43 in lysosomes. The data further indicate that ITCH interacts with and regulates Cx43 ubiquitination and that the ITCH-induced loss of Cx43-based gap junctions requires its catalytic HECT (homologous to E6-AP C-terminus) domain. The data also suggest that the ability of ITCH to efficiently promote loss of Cx43-based gap junctions and degradation of Cx43 depends on a functional PY (PPXY) motif in the C-terminal tail of Cx43. Together, these data provide new insights into the molecular basis underlying the degradation of Cx43 and have implications for the understanding of how intercellular communication via gap junctions is lost during cancer development.

• Langerud, Jonas; Eilertsen, Ina Andrassy; Moosavi, Hossein; Klokkerud, Solveig Margrete Knoph; Reims, Henrik Mikael & Backe, Ingeborg Flåten [Show all 13 contributors for this article] (2024). Multiregional transcriptomics identifies congruent consensus subtypes with prognostic value beyond tumor heterogeneity of colorectal cancer. Nature Communications. 15(1). doi: 10.1038/s41467-024-48706-2. Full text in Research Archive Show summary

  Intra-tumor heterogeneity compromises the clinical value of transcriptomic classifications of colorectal cancer. We investigated the prognostic effect of transcriptomic heterogeneity and the potential for classifications less vulnerable to heterogeneity in a single-hospital series of 1093 tumor samples from 692 patients, including multiregional samples from 98 primary tumors and 35 primary-metastasis sets. We show that intra-tumor heterogeneity of the consensus molecular subtypes (CMS) is frequent and has poor-prognostic associations independently of tumor microenvironment markers. Multiregional transcriptomics uncover cancer cell-intrinsic and low-heterogeneity signals that recapitulate the intrinsic CMSs proposed by single-cell sequencing. Further subclassification identifies congruent CMSs that explain a larger proportion of variation in patient survival than intra-tumor heterogeneity. Plasticity is indicated by discordant intrinsic phenotypes of matched primary and metastatic tumors. We conclude that multiregional sampling reconciles the prognostic power of tumor classifications from single-cell and bulk transcriptomics in the context of intra-tumor heterogeneity, and phenotypic plasticity challenges the reconciliation of primary and metastatic subtypes.

• Høland, Maren; Berg, Kaja Christine Graue; Eilertsen, Ina Andrassy; Bjerkehagen, Bodil; Kolberg, Matthias & Pedersen, Kjetil Boye [Show all 16 contributors for this article] (2023). Transcriptomic subtyping of malignant peripheral nerve sheath tumours highlights immune signatures, genomic profiles, patient survival and therapeutic targets. EBioMedicine. 97:104829, p. 1–17. doi: 10.1016/j.ebiom.2023.104829. Full text in Research Archive Show summary

  Background Malignant peripheral nerve sheath tumour (MPNST) is an aggressive orphan disease commonly affecting adolescents or young adults. Current knowledge of molecular tumour biology has been insufficient for development of rational treatment strategies. We aimed to discover molecular subtypes of potential clinical relevance. Methods Fresh frozen samples of MPNSTs (n = 94) and benign neurofibromas (n = 28) from 115 patients in a European multicentre study were analysed by DNA copy number and/or transcriptomic profiling. Unsupervised transcriptomic subtyping was performed and the subtypes characterized for genomic aberrations, clinicopathological associations and patient survival. Findings MPNSTs were classified into two transcriptomic subtypes defined primarily by immune signatures and proliferative processes. “Immune active” MPNSTs (44%) had sustained immune signals relative to neurofibromas, were more frequently low-grade (P = 0.01) and had favourable prognostic associations in a multivariable model of disease-specific survival with clinicopathological factors (hazard ratio 0.25, P = 0.003). “Immune deficient” MPNSTs were more aggressive and characterized by proliferative signatures, high genomic complexity, aberrant TP53 and PRC2 loss, as well as high relative expression of several potential actionable targets (EGFR, ERBB2, EZH2, KIF11, PLK1, RRM2). Integrated gene-wise analyses suggested a DNA copy number-basis for proliferative transcriptomic signatures in particular, and the tumour copy number burden further stratified the transcriptomic subtypes according to patient prognosis (P < 0.01). Interpretation Approximately half of MPNSTs belong to an “immune deficient” transcriptomic subtype associated with an aggressive disease course, PRC2 loss and expression of several potential therapeutic targets, providing a rationale for molecularly-guided intervention trials.

• Bogaard, Mari; Skotheim, Rolf I.; Maltau, Aase Margrethe Victoria; Kidd, Susanne Gundersen; Lothe, Ragnhild Adelheid & Axcrona, Karol [Show all 7 contributors for this article] (2023). "High proliferative cribriform prostate cancer" defines a patient subgroup with an inferior prognosis. Histopathology. ISSN 0309-0167. 83(6), p. 853–869. doi: 10.1111/his.15012. Full text in Research Archive Show summary

  Aims A cribriform pattern, reactive stroma (RS), PTEN, Ki67 and ERG are promising prognostic biomarkers in primary prostate cancer (PCa). We aim to determine the relative contribution of these factors and the Cancer of the Prostate Risk Assessment Postsurgical (CAPRA‐S) score in predicting PCa prognosis. Methods and results We included 475 patients who underwent radical prostatectomy (2010–12, median follow‐up = 8.7 years). Cribriform pattern was identified in 57% of patients, PTEN loss in 55%, ERG expression in 51%, RS in 39% and high Ki67 in 9%. In patients with multiple samples from the same malignant focus and either PTEN loss or high Ki67, intrafocal heterogeneity for PTEN and Ki67 expression was detected in 55% and 89%, respectively. In patients with samples from two or more foci, interfocal heterogeneity was detected in 46% for PTEN and 6% for Ki67. A cribriform pattern and Ki67 were independent predictors of biochemical recurrence (BCR) and clinical recurrence (CR), whereas ERG expression was an independent predictor of CR. Besides CAPRA‐S, a cribriform pattern provided the highest relative proportion of explained variation for predicting BCR (11%), and Ki67 provided the highest relative proportion of explained variation for CR (21%). In patients with a cribriform pattern, high Ki67 was associated with a higher risk of BCR [hazard ratio (HR) = 2.83, P  &lt; 0.001] and CR (HR = 4.35, P  &lt; 0.001). Conclusions High Ki67 in patients with a cribriform pattern identifies a patient subgroup with particularly poor prognosis, which we termed ‘high proliferative cribriform prostate cancer’. These results support reporting a cribriform pattern in pathology reports, and advocate implementing Ki67.

• Grasso, Chiara; Popovic, Maja; Isaevska, Elena; Lazzarato, Fulvio; Fiano, Valentina & Zugna, Daniela [Show all 47 contributors for this article] (2022). Association Study between Polymorphisms in DNA Methylation-Related Genes and Testicular Germ Cell Tumor Risk. Cancer Epidemiology, Biomarkers and Prevention. ISSN 1055-9965. 31(9), p. 1769–1779. doi: 10.1158/1055-9965.EPI-22-0123. Full text in Research Archive
• Carm, Kristina Totland; Johannessen, Bjarne; Bogaard, Mari; Bakken, Anne Cathrine; Maltau, Aase Margrethe Victoria & Hoff, Andreas Midbøe [Show all 10 contributors for this article] (2022). Somatic mutations reveal complex metastatic seeding from multifocal primary prostate cancer. International Journal of Cancer. ISSN 0020-7136. 152(5), p. 945–951. doi: 10.1002/ijc.34226. Full text in Research Archive Show summary

  Primary prostate cancer shows a striking intraorgan molecular heterogeneity, with multiple spatially separated malignant foci in the majority of patients. Metastatic prostate cancer, however, typically reveals more homogenous molecular profiles, suggesting a monoclonal origin of the metastatic lesions. Longitudinal mutational spectra, comparing multiple primary lesions with metastases from the same patients remain poorly defined. We have here analyzed somatic mutations in multisampled, spatio-temporal biobanked lesions (38 samples from primary foci and 1 sample from each of 8 metastases from seven prostate cancer patients) applying a custom-designed panel targeting 68 prostate cancer relevant genes. The metastatic samples were taken at time of primary surgery and up to 7 years later, and sampling included circulating tumor DNA in plasma or solid metastatic tissue samples. A total of 282 somatic mutations were detected, with a range of 0 to 25 mutations per sample. Although seven samples had solely private mutations, the remaining 39 samples had both private and shared mutations. Seventy-four percent of mutations in metastases were not found in any primary samples, and vice versa, 96% of mutations in primary cancers were not found in any metastatic samples. However, for three patients, shared mutations were found suggesting the focus of origin, including mutations in AKT1, FOXA1, HOXB13, RB1 and TP53. In conclusion, the spatio-temporal heterogeneous nature of multifocal disease is emphasized in our study, and underlines the importance of testing a recent sample in genomics-based precision medicine for metastatic prostate cancer.

• Bergsland, Christian Holst; Jeanmougin, Marine; Moosavi, Seyed Hossein; Svindland, Aud; Bruun, Jarle & Nesbakken, Arild [Show all 8 contributors for this article] (2022). Spatial analysis and CD25-expression identify regulatory T cells as predictors of a poor prognosis in colorectal cancer. Modern Pathology. ISSN 0893-3952. 35, p. 1236–1246. doi: 10.1038/s41379-022-01086-8. Full text in Research Archive Show summary

  Regulatory T cells (Tregs) are a heterogeneous cell population that can either suppress or stimulate immune responses. Tumor-infiltrating Tregs are associated with an adverse outcome from most cancer types, but have generally been found to be associated with a good prognosis in colorectal cancer (CRC). We investigated the prognostic heterogeneity of Tregs in CRC by co-expression patterns and spatial analyses with diverse T cell markers, using multiplex fluorescence immunohistochemistry and digital image analysis in two consecutive series of primary CRCs (total n = 1720). Treg infiltration in tumors, scored as FOXP3+ or CD4+/CD25+/FOXP3+ (triple-positive) cells, was strongly correlated to the overall amount of CD3+ and CD8+ T cells, and consequently associated with a favorable 5-year relapse-free survival rate among patients with stage I–III CRC who underwent complete tumor resection. However, high relative expression of the activation marker CD25 in triple-positive Tregs was independently associated with an adverse outcome in a multivariable model incorporating clinicopathological and known molecular prognostic markers (hazard ratio = 1.35, p = 0.028). Furthermore, spatial marker analysis based on Voronoi diagrams and permutation testing of cellular neighborhoods revealed a statistically significant proximity between Tregs and CD8+-cells in 18% of patients, and this was independently associated with a poor survival (multivariable hazard ratio = 1.36, p = 0.017). These results show prognostic heterogeneity of different Treg populations in primary CRC, and highlight the importance of multi-marker and spatial analyses for accurate immunophenotyping of tumors in relation to patient outcome.

• Strømme, Jonas Meier; Johannessen, Bjarne; Kidd, Susanne Gundersen; Bogaard, Mari; Carm, Kristina Totland & Zhang, Xiaokang [Show all 12 contributors for this article] (2022). Expressed prognostic biomarkers for primary prostate cancer independent of multifocality and transcriptome heterogeneity. Cancer Gene Therapy. ISSN 0929-1903. 29(8-9), p. 1276–1284. doi: 10.1038/s41417-022-00444-7. Full text in Research Archive Show summary

  The majority of prostate cancer patients are diagnosed with multiple primary malignant foci. The distinct foci are exceptionally heterogeneous with regard to DNA mutations, but whether this is recapitulated at the transcriptome level remains unknown. In this study, inter- and intrafocal heterogeneity has been assessed by whole-transcriptome sequencing of 87 tissue samples from 23 patients with localized prostate cancer treated with radical prostatectomy. From each patient, multiple samples were taken from one or more malignant foci, in addition to one sample from benign prostate tissue. Transcriptomic profiles of different malignant foci from the same patient showed a similar level of heterogeneity as tumors from different patients. This applies to expression of genes, fusion genes, and somatic mutations. Within-patient pair-wise analyses identified expression patterns linked to ETS status and extraprostatic extension. A set of 62 genes were found with low intrapatient heterogeneity and high interpatient heterogeneity, retaining stable expression profiles across foci within the same patient. Among these, 16 genes are associated with biochemical recurrence in a separately published study and are therefore nominated as biomarkers with prognostic value regardless of which malignant focus is sampled. In conclusion, an extensive heterogeneity in multifocal prostate cancer is confirmed at the gene expression level. Diagnostic biomarkers were identified for ETS positive samples and samples from extraprostatic extensions. Finally, prognostic biomarkers independent of multifocal heterogeneity were found.

• Aasebø, Kristine Øverås; Bruun, Jarle; Bergsland, Christian Holst; Nunes, Luis; Eide, Geir Egil & Pfeiffer, Per [Show all 10 contributors for this article] (2022). Prognostic role of tumour infiltrating lymphocytes and macrophages in relation to tumour MSI, CDX2 and BRAF status: a population-based study of metastatic colorectal cancer patients. British Journal of Cancer. ISSN 0007-0920. 126(1), p. 48–56. doi: 10.1038/s41416-021-01586-5. Full text in Research Archive
• Kidd, Susanne Gundersen; Bogaard, Mari; Carm, Kristina Totland; Bakken, Anne Cathrine; Maltau, Aase Margrethe Victoria & Løvf, Marthe [Show all 10 contributors for this article] (2022). In situ expression of ERG protein in the context of tumor heterogeneity identifies prostate cancer patients with inferior prognosis. Molecular Oncology. ISSN 1574-7891. 16(15), p. 2810–2822. doi: 10.1002/1878-0261.13225. Full text in Research Archive Show summary

  Prognostic biomarkers for prostate cancer are needed to improve prediction of disease course and guide treatment decisions. However, biomarker development is complicated by the common multifocality and heterogeneity of the disease. We aimed to determine the prognostic value of candidate biomarkers transcriptional regulator ERG and related ETS family genes, while considering tumor heterogeneity. In a multisampled, prospective, and treatment-naïve radical prostatectomy cohort from one tertiary center (2010–2012, median follow-up 8.1 years), we analyzed ERG protein (480 patients; 2047 tissue cores), and RNA of several ETS genes in a subcohort (165 patients; 778 fresh-frozen tissue samples). Intra- and interfocal heterogeneity was identified in 29% and 33% (ERG protein) and 39% and 27% (ETS RNA) of patients, respectively. ERG protein and ETS RNA was identified exclusively in a nonindex tumor in 31% and 32% of patients, respectively. ERG protein demonstrated independent prognostic value in predicting biochemical (P = 0.04) and clinical recurrence (P = 0.004) and appeared to have greatest prognostic value for patients with Grade Groups 4–5. In conclusion, when heterogeneity is considered, ERG protein is a robust prognostic biomarker for prostate cancer.

• Sveen, Anita; Johannessen, Bjarne; Eilertsen, Ina Andrassy; Røsok, Bård Ingvald; Gulla, Marie & Eide, Peter Andreas Wold [Show all 14 contributors for this article] (2021). The expressed mutational landscape of microsatellite stable colorectal cancers. Genome Medicine. 13. doi: 10.1186/s13073-021-00955-2. Full text in Research Archive Show summary

  Background Colorectal cancer is the 2nd leading cause of cancer-related deaths with few patients benefiting from biomarker-guided therapy. Mutation expression is essential for accurate interpretation of mutations as biomarkers, but surprisingly, little has been done to analyze somatic cancer mutations on the expression level. We report a large-scale analysis of allele-specific mutation expression. Methods Whole-exome and total RNA sequencing was performed on 137 samples from 121 microsatellite stable colorectal cancers, including multiregional samples of primary and metastatic tumors from 4 patients. Data were integrated with allele-specific resolution. Results were validated in an independent set of 241 colon cancers. Therapeutic associations were explored by pharmacogenomic profiling of 15 cell lines or patient-derived organoids. Results The median proportion of expressed mutations per tumor was 34%. Cancer-critical mutations had the highest expression frequency (gene-wise mean of 58%), independent of frequent allelic imbalance. Systematic deviation from the general pattern of expression levels according to allelic frequencies was detected, including preferential expression of mutated alleles dependent on the mutation type and target gene. Translational relevance was suggested by correlations of KRAS/NRAS or TP53 mutation expression levels with downstream oncogenic signatures (p < 0.03), overall survival among patients with stage II and III cancer (KRAS/NRAS: hazard ratio 6.1, p = 0.0070), and targeted drug sensitivity. The latter was demonstrated for EGFR and MDM2 inhibition in pre-clinical models. Conclusions Only a subset of mutations in microsatellite stable colorectal cancers were expressed, and the “expressed mutation dose” may provide an opportunity for more fine-tuned biomarker interpretations.

• Eide, Peter Andreas Wold; Moosavi, Seyed Hossein; Eilertsen, Ina Andrassy; Brunsell, Tuva Høst; Langerud, Jonas & Berg, Kaja Christine Graue [Show all 11 contributors for this article] (2021). Metastatic heterogeneity of the consensus molecular subtypes of colorectal cancer. NPJ GENOMIC MEDICINE. 6:59, p. 1–9. doi: 10.1038/s41525-021-00223-7. Full text in Research Archive Show summary

  Abstract Gene expression-based subtypes of colorectal cancer have clinical relevance, but the representativeness of primary tumors and the consensus molecular subtypes (CMS) for metastatic cancers is not well known. We investigated the metastatic heterogeneity of CMS. The best approach to subtype translation was delineated by comparisons of transcriptomic profiles from 317 primary tumors and 295 liver metastases, including multi-metastatic samples from 45 patients and 14 primary-metastasis sets. Associations were validated in an external data set ( n  = 618). Projection of metastases onto principal components of primary tumors showed that metastases were depleted of CMS1-immune/CMS3-metabolic signals, enriched for CMS4-mesenchymal/stromal signals, and heavily influenced by the microenvironment. The tailored CMS classifier (available in an updated version of the R package CMScaller) therefore implemented an approach to regress out the liver tissue background. The majority of classified metastases were either CMS2 or CMS4. Nonetheless, subtype switching and inter-metastatic CMS heterogeneity were frequent and increased with sampling intensity. Poor-prognostic value of CMS1/3 metastases was consistent in the context of intra-patient tumor heterogeneity.

• Kryeziu, Kushtrim; Moosavi, Seyed Hossein; Bergsland, Christian Holst; Guren, Marianne; Eide, Peter Andreas Wold & Totland, Max [Show all 11 contributors for this article] (2021). Increased sensitivity to SMAC mimetic LCL161 identified by longitudinal ex vivo pharmacogenomics of recurrent, KRAS mutated rectal cancer liver metastases. Journal of Translational Medicine. 19:384, p. 1–11. doi: 10.1186/s12967-021-03062-3. Full text in Research Archive Show summary

  Tumor heterogeneity is a primary cause of treatment failure. However, changes in drug sensitivity over time are not well mapped in cancer. Patient-derived organoids (PDOs) may predict clinical drug responses ex vivo and ofer an opportunity to evaluate novel treatment strategies in a personalized fashion. Here we have evaluated spatio-temporal functional and molecular dynamics of fve PDO models established after hepatic re-resections and neoadjuvant combination chemotherapies in a patient with microsatellite stable and KRAS mutated metastatic rectal cancer. Histopathological diferentiation phenotypes of the PDOs corresponded with the liver metastases, and ex vivo drug sensitivities generally refected clinical responses and selection pressure, assessed in comparison to a reference data set of PDOs from metastatic colorectal cancers. PDOs from the initial versus the two recurrent metastatic settings showed heterogeneous cell morphologies, protein marker expression, and drug sensitivities. Exploratory analyses of a drug screen library of 33 investigational anticancer agents showed the strongest ex vivo sensitivity to the SMAC mimetic LCL161 in PDOs of recurrent disease compared to those of the initial metastasis. Functional analyses confrmed target inhibition and apoptosis induction in the LCL161 sensitive PDOs from the recurrent metastases. Gene expression analyses indicated an association between LCL161 sensitivity and tumor necrosis factor alpha signaling and RIPK1 gene expression. In conclusion, LCL161 was identifed as a possible experimental therapy of a metastatic rectal cancer that relapsed after hepatic resection and standard systemic treatment.

• Bruun, Jarle; Eide, Peter Andreas Wold; Bergsland, Christian Holst; Brück, Oscar; Svindland, Aud & Arjama, Mariliina [Show all 13 contributors for this article] (2021). E-cadherin is a robust prognostic biomarker in colorectal cancer and low expression is associated with sensitivity to inhibitors of topoisomerase, aurora, and HSP90 in preclinical models. Molecular Oncology. ISSN 1574-7891. 00, p. 1–18. doi: 10.1002/1878-0261.13159. Full text in Research Archive Show summary

  Cell–cell and cell–matrix adhesion proteins that have been implicated in colorectal epithelial integrity and epithelial-to-mesenchymal transition could be robust prognostic and potential predictive biomarkers for standard and novel therapies. We analyzed in situ protein expression of E-cadherin (ECAD), integrin β4 (ITGB4), zonula occludens 1 (ZO-1), and cytokeratins in a single-hospital series of Norwegian patients with colorectal cancer (CRC) stages I–IV (n = 922) using multiplex fluorescence-based immunohistochemistry (mfIHC) on tissue microarrays. Pharmacoproteomic associations were explored in 35 CRC cell lines annotated with drug sensitivity data on > 400 approved and investigational drugs. ECAD, ITGB4, and ZO-1 were positively associated with survival, while cytokeratins were negatively associated with survival. Only ECAD showed independent prognostic value in multivariable Cox models. Clinical and molecular associations for ECAD were technically validated on a different mfIHC platform, and the prognostic value was validated in another Norwegian series (n = 798). In preclinical models, low and high ECAD expression differentially associated with sensitivity to topoisomerase, aurora, and HSP90 inhibitors, and EGFR inhibitors. E-cadherin protein expression is a robust prognostic biomarker with potential clinical utility in CRC.

• Moosavi, Seyed Hossein; Eide, Peter Andreas Wold; Eilertsen, Ina Andrassy; Brunsell, Tuva Høst; Berg, Kaja Christine Graue & Røsok, Bård Ingvald [Show all 12 contributors for this article] (2021). De novo transcriptomic subtyping of colorectal cancer liver metastases in the context of tumor heterogeneity. Genome Medicine. 13, p. 143–143. doi: 10.1186/s13073-021-00956-1. Full text in Research Archive Show summary

  Background Gene expression-based subtyping has the potential to form a new paradigm for stratified treatment of colorectal cancer. However, current frameworks are based on the transcriptomic profiles of primary tumors, and metastatic heterogeneity is a challenge. Here we aimed to develop a de novo metastasis-oriented framework. Methods In total, 829 transcriptomic profiles from patients with colorectal cancer were analyzed, including primary tumors, liver metastases, and non-malignant liver samples. High-resolution microarray gene expression profiling was performed of 283 liver metastases from 171 patients treated by hepatic resection, including multiregional and/or multi-metastatic samples from each of 47 patients. A single randomly selected liver metastasis sample from each patient was used for unsupervised subtype discovery by nonnegative matrix factorization, and a random forest prediction model was trained to classify multi-metastatic samples, as well as liver metastases from two independent series of 308 additional patients. Results Initial comparisons with non-malignant liver samples and primary colorectal tumors showed a highly variable degree of influence from the liver microenvironment in metastases, which contributed to inter-metastatic transcriptomic heterogeneity, but did not define subtype distinctions. The de novo liver metastasis subtype (LMS) framework recapitulated the main distinction between epithelial-like and mesenchymal-like tumors, with a strong immune and stromal component only in the latter. We also identified biologically distinct epithelial-like subtypes originating from different progenitor cell types. LMS1 metastases had several transcriptomic features of cancer aggressiveness, including secretory progenitor cell origin, oncogenic addictions, and microsatellite instability in a microsatellite stable background, as well as frequent RAS/TP53 co-mutations. The poor-prognostic association of LMS1 metastases was independent of mutation status, clinicopathological variables, and current subtyping frameworks (consensus molecular subtypes and colorectal cancer intrinsic subtypes). LMS1 was also the least heterogeneous subtype in comparisons of multiple metastases per patient, and tumor heterogeneity did not confound the prognostic value of LMS1. Conclusions We report the first large study of multi-metastatic gene expression profiling of colorectal cancer. The new metastasis-oriented subtyping framework showed potential for clinically relevant transcriptomic classification in the context of metastatic heterogeneity, and an LMS1 mini-classifier was constructed to facilitate prognostic stratification and further clinical testing.

• Pluta, John; Pyle, Louise C.; Nead, Kevin T.; Wilf, Rona; Li, Mingyao & Mitra, Nandita [Show all 57 contributors for this article] (2021). Identification of 22 susceptibility loci associated with testicular germ cell tumors. Nature Communications. 12(1), p. 1–13. doi: 10.1038/s41467-021-24334-y. Full text in Research Archive
• Kidd, Susanne Gundersen; Carm, Kristina Totland; Bogaard, Mari; Olsen, Linn Guro; Bakken, Anne Cathrine & Løvf, Marthe [Show all 10 contributors for this article] (2021). High expression of SCHLAP1 in primary prostate cancer is an independent predictor of biochemical recurrence, despite substantial heterogeneity. Neoplasia. ISSN 1522-8002. 23(6), p. 634–641. doi: 10.1016/j.neo.2021.05.012. Full text in Research Archive Show summary

  In primary prostate cancer, the common multifocality and heterogeneity are major obstacles in finding robust prognostic tissue biomarkers. The long noncoding RNA SCHLAP1 has been suggested, but its prognostic value has not been investigated in the context of tumor heterogeneity. In the present study, expression of SCHLAP1 was investigated using real-time RT-PCR in a multisampled series of 778 tissue samples from radical prostatectomies of 164 prostate cancer patients (median follow-up time 7.4 y). The prognostic value of SCHLAP1 was evaluated with biochemical recurrence as endpoint. In total, 29% of patients were classified as having high expression of SCHLAP1 in at least one malignant sample. Among these, inter- and intrafocal heterogeneity was detected in 72% and 56%, respectively. High expression of SCHLAP1 was shown to be a predictor of biochemical recurrence in both uni- and multivariable cox regression analyses ( P < 0.001 and P = 0.02). High expression of SCHLAP1 was also significantly associated with adverse clinicopathological characteristics, including grade group, high pT stage, invasive cribriform growth/intraductal carcinoma of the prostate, and reactive stroma. In conclusion, high expression of SCHLAP1 in at least one malignant sample is a robust prognostic biomarker in primary prostate cancer. For the first time, high SCHLAP1 expression has been associated with the aggressive histopathologic feature reactive stroma. The expression of SCHLAP1 is highly heterogeneous, and analysis of multiple samples is therefore crucial in determination of the SCHLAP1 status of a patient.

• Slørdahl, Kathinka; Klotz, Dagmar; Olsen, Jan Åge; Skovlund, Eva; Undseth, Christine & Abildgaard, Heidi L. [Show all 17 contributors for this article] (2021). Treatment outcomes and prognostic factors after chemoradiotherapy for anal cancer. Acta Oncologica. ISSN 0284-186X. 60(7), p. 921–930. doi: 10.1080/0284186X.2021.1918763. Full text in Research Archive Show summary

  Background Squamous cell carcinoma of the anus (SCCA) is a rare malignancy with rising incidence, associated with human papilloma virus (HPV). Chemoradiotherapy (CRT) is the preferred treatment. The purpose was to investigate treatment failure, survival and prognostic factors after CRT. Material and methods In this prospective observational study from a large regional centre, 141 patients were included from 2013 to 2017, and 132 were eligible for analysis. The main inclusion criteria were SCCA, planned radiotherapy, and performance status (ECOG) ≤2. Patient characteristics, disease stage, treatment, and treatment response were prospectively registered. Disease-free survival (DFS), overall survival (OS), and locoregional treatment failure after CRT were analysed. Hazard ratios (HRs) were estimated with Cox`s proportional hazards model. Results Median follow-up was 54 (range 6–71) months. Eighteen patients (14%) had treatment failures after CRT; of these 10 (8%) had residual tumour, and 8 (6%) relapse as first failure. The first treatment failure was locoregional (11 patients), distant (5 patients), and both (2 patients). Salvage abdomino-perineal resection was performed in 10 patients, 2 had resections of metastases, and 3 both. DFS was 85% at 3 years and 78% at 5 years. OS was 93% at 3 years and 86% at 5 years. In analyses adjusted for age and gender, HPV negative tumours (HR 2.5, p = 0.024), N3 disease (HR 2.6, p = 0.024), and tumour size ≥4 cm (HR 2.4, p = 0.038) were negative prognostic factors for DFS. Conclusion State-of-the-art chemoradiotherapy for SCCA resulted in excellent outcomes, and improved survival compared with previous national data, with <15% treatment failures and a 3-year DFS of >80%.

• Wise, Jillian; Nakken, Sigve; Steen, Chloe Beate; Vodak, Daniel; Trøen, Gunhild & Johannessen, Bjarne [Show all 24 contributors for this article] (2020). Mutational dynamics and immune evasion in diffuse large B-cell lymphoma explored in a relapse-enriched patient series. Blood Advances. ISSN 2473-9529. 4(9), p. 1859–1866. doi: 10.1182/bloodadvances.2019001325. Full text in Research Archive Show summary

  • Diagnostic and relapse diffuse large B-cell lymphoma (DLBCL) biopsies reveal increased mutational burden/loss of heterozygosity in HLA-A. • Serially sampled tumor biopsies provide insight into therapeutic targets and evolutionary divergence in relapsed/refractory DLBCL.

• Smeby, Jørgen; Kryeziu, Kushtrim; Berg, Kaja Christine Graue; Eilertsen, Ina Andrassy; Eide, Peter Andreas Wold & Johannessen, Bjarne [Show all 11 contributors for this article] (2020). Molecular correlates of sensitivity to PARP inhibition beyond homologous recombination deficiency in pre-clinical models of colorectal cancer point to wild-type TP53 activity. EBioMedicine. 59:102923, p. 1–13. doi: 10.1016/j.ebiom.2020.102923. Full text in Research Archive Show summary

  Background PARP inhibitors are active in various tumour types beyond BRCA-mutant cancers, but their activity and molecular correlates in colorectal cancer (CRC) are not well studied. Methods Mutations and genome-wide mutational patterns associated with homologous recombination deficiency (HRD) were investigated in 255 primary CRCs with whole-exome sequencing and/or DNA copy number data. Efficacy of five PARP inhibitors and their molecular correlates were evaluated in 93 CRC cell lines partly annotated with mutational-, DNA copy number-, and/or gene expression profiles. Post-treatment gene expression profiling and specific protein expression analyses were performed in two pairs of PARP inhibitor sensitive and resistant cell lines. Findings A subset of microsatellite stable (MSS) CRCs had truncating mutations in homologous recombination-related genes, but these were not associated with genomic signatures of HRD. Eight CRC cell lines (9%) were sensitive to PARP inhibition, but sensitivity was not predicted by HRD-related genomic and transcriptomic signatures. In contrast, drug sensitivity in MSS cell lines was strongly associated with TP53 wild-type status (odds ratio 15.7, p = 0.023) and TP53-related expression signatures. Increased downstream TP53 activity was among the primary response mechanisms, and TP53 inhibition antagonized the effect of PARP inhibitors. Wild-type TP53-mediated suppression of RAD51 was identified as a possible mechanism of action for sensitivity to PARP inhibition. Interpretation PARP inhibitors are active in a subset of CRC cell lines and preserved TP53 function may increase the likelihood of response.

• Berg, Kaja Christine Graue; Brunsell, Tuva Høst; Sveen, Anita; Alagaratnam, Sharmini; Bjørnslett, Merete Pauline & Hektoen, Merete [Show all 11 contributors for this article] (2020). Genomic and prognostic heterogeneity among RAS/BRAFV600E/TP53 co-mutated resectable colorectal liver metastases. Molecular Oncology. ISSN 1574-7891. p. 1–16. doi: 10.1002/1878-0261.12885. Full text in Research Archive Show summary

  Hepatic resection is potentially curative for patients with colorectal liver metastases, but the treatment benefit varies. KRAS/NRAS (RAS)/TP53 co‐mutations are associated with a poor prognosis after resection, but there is large variation in patient outcome within the mutation groups, and genetic testing is currently not used to evaluate benefit from surgery. We have investigated the potential for improved prognostic stratification by combined biomarker analysis with DNA copy number aberrations (CNAs), and taking tumor heterogeneity into account. We determined the mutation status of RAS, BRAFV600, and TP53 in 441 liver lesions from 171 patients treated by partial hepatectomy for metastatic colorectal cancer. CNAs were profiled in 232 tumors from 67 of the patients. Mutations and high‐level amplifications of cancer‐critical genes, the latter including ERBB2 and EGFR, were predominantly homogeneous within patients. RAS/BRAFV600E and TP53 co‐mutations were associated with a poor patient outcome (hazard ratio, HR, 3.9, 95% confidence interval, CI, 1.3–11.1, P = 0.012) in multivariable analyses with clinicopathological variables. The genome‐wide CNA burden and intrapatient intermetastatic CNA heterogeneity varied within the mutation groups, and the CNA burden had prognostic associations in univariable analysis. Combined prognostic analyses of RAS/BRAFV600E/TP53 mutations and CNAs, either as a high CNA burden or high intermetastatic CNA heterogeneity, identified patients with a particularly poor outcome (co‐mutation/high CNA burden: HR 2.7, 95% CI 1.2–5.9, P = 0.013; co‐mutation/high CNA heterogeneity: HR 2.5, 95% CI 1.1–5.6, P = 0.022). In conclusion, DNA copy number profiling identified genomic and prognostic heterogeneity among patients with resectable colorectal liver metastases with co‐mutated RAS/BRAFV600E/TP53.

• Bergsland, Christian Holst; Bruun, Jarle; Guren, Marianne; Svindland, Aud; Bjørnslett, Merete Pauline & Smeby, Jørgen [Show all 16 contributors for this article] (2020). Prediction of relapse-free survival according to adjuvant chemotherapy and regulator of chromosome condensation 2 (RCC2) expression in colorectal cancer. ESMO Open. 5:e001040(6), p. 1–12. doi: 10.1136/esmoopen-2020-001040. Full text in Research Archive Show summary

  Background There is a need for improved selection of patients for adjuvant chemotherapy after resection of non-metastatic colorectal cancer (CRC). Regulator of chromosome condensation 2 (RCC2) is a potential prognostic biomarker. We report on the establishment of a robust protocol for RCC2 expression analysis and prognostic tumour biomarker evaluation in patients who did and did not receive adjuvant chemotherapy. Materials and methods RCC2 was analysed in 2916 primary CRCs from the QUASAR2 randomised trial and two single-hospital Norwegian series. A new protocol using fluorescent antibody staining and digital image analysis was optimised. Biomarker value for 5-year relapse-free survival was analysed in relation to tumour stage, adjuvant chemotherapy and the molecular markers microsatellite instability, KRAS/BRAFV600E/TP53 mutations and CDX2 expression. Results Low RCC2 expression was scored in 41% of 2696 evaluable samples. Among patients with stage I–III CRC who had not received adjuvant chemotherapy, low RCC2 expression was an independent marker of inferior 5-year relapse-free survival in multivariable Cox models including clinicopathological factors and molecular markers (HR 1.45, 95% CI 1.09 to 1.94, p=0.012, N=521). RCC2 was not prognostic in patients who had received adjuvant chemotherapy, neither in QUASAR2 nor the pooled Norwegian series. The interaction between RCC2 and adjuvant chemotherapy for prediction of patient outcome was significant in stage III, and strongest among patients with microsatellite stable tumours (pinteraction=0.028). Conclusions Low expression of RCC2 is a biomarker for poor prognosis in patients with stage I–III CRC and seems to be a predictive biomarker for effect of adjuvant chemotherapy.

• Flatin, Bjørnar Tovson Bae; Vedeld, Hege Marie; Pinto, Rita; Langerud, Jonas; Lind, Guro Elisabeth & Lothe, Ragnhild A [Show all 8 contributors for this article] (2020). Multiregional assessment of CIMP in primary colorectal cancers: Phenotype concordance but marker variability. International Journal of Cancer. ISSN 0020-7136. p. 1–6. doi: 10.1002/ijc.33425. Full text in Research Archive Show summary

  Intratumor heterogeneity of colorectal cancers (CRCs) is manifested both at the genomic and epigenomic levels. Early genetic aberrations in carcinogenesis are clonal and present throughout the tumors, but less is known about the heterogeneity of the epigenetic CpG island methylator phenotype (CIMP). CIMP characterizes a subgroup of CRCs thought to originate from specific precursor lesions, and it is defined by widespread DNA methylation within promoter regions. In this work, we investigated CIMP in two to four multiregional samples from 30 primary tumors (n = 86 samples) using the consensus Weisenberger gene panel (CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1). Twenty‐nine of 30 tumors (97%) showed concordant CIMP status in all samples, and percent methylated reference (PMR) values of all five markers had higher intertumor than intratumor variation (P value = 1.5e−09). However, a third of the CIMP+ tumors exhibited discrepancies in methylation status in at least one of the five gene markers. To conclude, CIMP status was consistent within primary CRCs, and it is likely a clonal phenotype. However, spatial discordances of the individual genes suggest that large‐scale analysis of multiregional samples could be of interest for identifying CIMP markers that are robust to intratumor heterogeneity.

• Hoff, Andreas Midbøe; Kraggerud, Sigrid M.; Alagaratnam, Sharmini; Berg, Kaja Christine Graue; Johannessen, Bjarne & Høland, Maren [Show all 11 contributors for this article] (2020). Frequent copy number gains of SLC2A3 and ETV1 in testicular embryonal carcinomas. Endocrine-Related Cancer. ISSN 1351-0088. 27(9), p. 457–468. doi: 10.1530/ERC-20-0064. Full text in Research Archive Show summary

  Testicular germ cell tumours (TGCTs) appear as different histological subtypes or mixtures of these. They show similar, multiple DNA copy number changes, where gain of 12p is pathognomonic. However, few high-resolution analyses have been performed and focal DNA copy number changes with corresponding candidate target genes remain poorly described for individual subtypes. We present the first high-resolution DNA copy number aberration (CNA) analysis on the subtype embryonal carcinomas (ECs), including 13 primary ECs and 5 EC cell lines. We identified recurrent gains and losses and allele-specific CNAs. Within these regions, we nominate 30 genes that may be of interest to the EC subtype. By in silico analysis of data from 150 TGCTs from The Cancer Genome Atlas (TCGA), we further investigated CNAs, RNA expression, somatic mutations and fusion transcripts of these genes. Among primary ECs, ploidy ranged between 2.3 and 5.0, and the most common aberrations were DNA copy number gains at chromosome (arm) 7, 8, 12p, and 17, losses at 4, 10, 11, and 18, replicating known TGCT genome characteristics. Gain of whole or parts of 12p was found in all samples, including a highly amplified 100 kbp segment at 12p13.31, containing SLC2A3. Gain at 7p21, encompassing ETV1 , was the second most frequent aberration. In conclusion, we present novel CNAs and the genes located within these regions, where the copy number gain of SLC2A3 and ETV1 are of interest, and which copy number levels also correlate with expression in TGCTs.

• Jeanmougin, Marine; Håvik, Annette Bentsen; Cekaite, Lina; Brandal, Petter; Sveen, Anita & Meling, Torstein Ragnar [Show all 11 contributors for this article] (2020). Improved prognostication of glioblastoma beyond molecular subtyping by transcriptional profiling of the tumor microenvironment. Molecular Oncology. ISSN 1574-7891. p. 1016–1027. doi: 10.1002/1878-0261.12668. Full text in Research Archive Show summary

  Glioblastoma (GBM), the most aggressive form of brain cancer, is characterized by a high level of molecular heterogeneity, and infiltration by various immune and stromal cell populations. Important advances have been made in deciphering the microenvironment of GBMs, but its association with existing molecular subtypes and its potential prognostic role remain elusive. We have investigated the abundance of infiltrating immune and stromal cells in silico, from gene expression profiles. Two cohorts, including in‐house normal brain and glioma samples (n = 70) and a large sample set from TCGA (n = 393), were combined into a single exploratory dataset. A third independent cohort (n = 124) was used for validation. Tumors were clustered based on their microenvironment infiltration profiles, and associations with known GBM molecular subtypes and patient outcome were tested a posteriori in a multivariable setting. We identified a subset of GBM samples with significantly higher abundances of most immune and stromal cell populations. This subset showed increased expression of both immune suppressor and immune effector genes compared to other GBMs and was enriched for the mesenchymal molecular subtype. Survival analyses suggested that tumor microenvironment infiltration pattern was an independent prognostic factor for GBM patients. Among all, patients with the mesenchymal subtype with low immune and stromal infiltration had the poorest survival. By combining molecular subtyping with gene expression measures of tumor infiltration, the present work contributes with improving prognostic models in GBM.

• Lopes, Nair Susana da Costa Florim Ribeiro; Bergsland, Christian Holst; Bruun, Jarle; Bjørnslett, Merete Pauline; Vieira, André Filipe & Mesquita, Patrícia [Show all 14 contributors for this article] (2020). A panel of intestinal differentiation markers (CDX2, GPA33, and LI-cadherin) identifies gastric cancer patients with favourable prognosis. Gastric Cancer. ISSN 1436-3291. 23(5), p. 811–823. doi: 10.1007/s10120-020-01064-6. Full text in Research Archive
• Bruun, Jarle; Kryeziu, Kushtrim; Eide, Peter Andreas Wold; Moosavi, Seyed Hossein; Eilertsen, Ina Andrassy & Røsok, Bård Ingvald [Show all 21 contributors for this article] (2020). Patient-Derived Organoids from Multiple Colorectal Cancer Liver Metastases Reveal Moderate Intra-patient Pharmacotranscriptomic Heterogeneity. Clinical Cancer Research. ISSN 1078-0432. 26(15), p. 4107–4119. doi: 10.1158/1078-0432.CCR-19-3637. Full text in Research Archive Show summary

  Abstract Purpose: Molecular tumor heterogeneity may have important implications for the efficacy of targeted therapies in metastatic cancers. Inter-metastatic heterogeneity of sensitivity to anticancer agents has not been well explored in colorectal cancer. Experimental Design: We established a platform for ex vivo pharmacogenomic profiling of patient-derived organoids (PDO) from resected colorectal cancer liver metastases. Drug sensitivity testing (n = 40 clinically relevant agents) and gene expression profiling were performed on 39 metastases from 22 patients. Results: Three drug–response clusters were identified among the colorectal cancer metastases, based primarily on sensitivities to EGFR and/or MDM2 inhibition, and corresponding with RAS mutations and TP53 activity. Potentially effective therapies, including off-label use of drugs approved for other cancer types, could be nominated for eighteen patients (82%). Antimetabolites and targeted agents lacking a decisive genomic marker had stronger differential activity than most approved chemotherapies. We found limited intra-patient drug sensitivity heterogeneity between PDOs from multiple (2–5) liver metastases from each of ten patients. This was recapitulated at the gene expression level, with a highly proportional degree of transcriptomic and pharmacological variation. One PDO with a multi-drug resistance profile, including resistance to EGFR inhibition in a RAS-mutant background, showed sensitivity to MEK plus mTOR/AKT inhibition, corresponding with low-level PTEN expression. Conclusions: Intra-patient inter-metastatic pharmacological heterogeneity was not pronounced and ex vivo drug screening may identify novel treatment options for metastatic colorectal cancer. Variation in drug sensitivities was reflected at the transcriptomic level, suggesting potential to develop gene expression–based predictive signatures to guide experimental therapies.

• Glaire, Mark A; Domingo, Enric; Sveen, Anita; Bruun, Jarle; Nesbakken, Arild & Nicholson, George [Show all 16 contributors for this article] (2019). Tumour-infiltrating CD8+ lymphocytes and colorectal cancer recurrence by tumour and nodal stage. British Journal of Cancer. ISSN 0007-0920. 121, p. 474–482. doi: 10.1038/s41416-019-0540-4. Full text in Research Archive
• Brunsell, Tuva Høst; Sveen, Anita; Bjørnbeth, Bjørn Atle; Røsok, Bård Ingvald; Danielsen, Stine Aske & Brudvik, Kristoffer Watten [Show all 13 contributors for this article] (2019). High Concordance and Negative Prognostic Impact of RAS/BRAF/PIK3CA Mutations in Multiple Resected Colorectal Liver Metastases. Clinical colorectal cancer. ISSN 1533-0028. 19(1), p. 26–47. doi: 10.1016/j.clcc.2019.09.003. Full text in Research Archive Show summary

  Background The prevalence and clinical implications of genetic heterogeneity in patients with multiple colorectal liver metastases remain largely unknown. In a prospective series of patients undergoing resection of colorectal liver metastases, the aim was to investigate the inter-metastatic and primary-to-metastatic heterogeneity of mutations in KRAS, NRAS, BRAF, and PIK3CA and their prognostic impact. Patients and Methods We analyzed the mutation status among 372 liver metastases and 78 primary tumors from 106 patients by methods used in clinical routine testing, by Sanger sequencing, by next-generation sequencing (NGS), and/or by droplet digital polymerase chain reaction. The 3-year cancer-specific survival (CSS) was analyzed using the Kaplan-Meier method. Results Although Sanger sequencing indicated inter-metastatic mutation heterogeneity in 14 of 97 patients (14%), almost all cases were refuted by high-sensitive NGS. Also, heterogeneity among metastatic deposits was concluded only for PIK3CA in 2 patients. Similarly, primary-to-metastatic heterogeneity was indicated in 8 of 78 patients (10%) using Sanger sequencing but for only 2 patients after NGS, showing the emergence of 1 KRAS and 1 PIK3CA mutation in the metastatic lesions. KRAS mutations were present in 53 of 106 patients (50%) and were associated with poorer 3-year CSS after liver resection (37% vs. 61% for KRAS wild-type; P = .004). Poor prognostic associations were found also for the combination of KRAS/NRAS/BRAF mutations compared with triple wild-type (P = .002). Conclusion Intra-patient mutation heterogeneity was virtually undetected, both between the primary tumor and the liver metastases and among the metastatic deposits. KRAS mutations separately, and KRAS/NRAS/BRAF mutations combined, were associated with poor patient survival after partial liver resection.

• Eilertsen, Ina Andrassy; Moosavi, Seyed Hossein; Strømme, Jonas Meier; Nesbakken, Arild; Johannessen, Bjarne & Lothe, Ragnhild A [Show all 7 contributors for this article] (2019). Technical differences between sequencing and microarray platforms impact transcriptomic subtyping of colorectal cancer. Cancer Letters. ISSN 0304-3835. 469, p. 246–255. doi: 10.1016/j.canlet.2019.10.040. Full text in Research Archive Show summary

  Gene expression profiling has increasing relevance in the molecular screening of patients with colorectal cancer (CRC). We investigated potential platform-specific effects on transcriptomic subtyping according to established frameworks by comparisons of expression profiles from RNA sequencing and exon-resolution microarrays in 126 primary microsatellite stable CRCs. There was a strong platform correspondence in global gene expression levels, albeit with systematic technical bias likely attributed to few sequencing reads covering short (<2000 nucleotides) and/or lowly expressed genes (<1 FPKM), as well as over-saturation of highly expressed genes on microarrays. Classification concordances according to both the consensus molecular subtypes and CRC intrinsic subtypes (CRIS) were also strong, but with disproportionate subtype distributions between platforms caused by frequent disagreements in adherence to sample classification thresholds. Subtypes defined largely by genes expressed at low levels, including the CRIS-D subtype and the estimated level of tumor-infiltrating cytotoxic lymphocytes, had a weaker correspondence in classification metrics between platforms. In conclusion, even subtle differences between platforms suggest that clinical translation of transcriptomic CRC subtyping frameworks is dependent on assay standardization, and systematic technical biases reinforce the need for careful selection of classifier genes.

• Smeby, Jørgen; Sveen, Anita; Bergsland, Christian Holst; Eilertsen, Ina Andrassy; Danielsen, Stine Aske & Eide, Peter Andreas Wold [Show all 11 contributors for this article] (2019). Exploratory analyses of consensus molecular subtype-dependent associations of TP53 mutations with immunomodulation and prognosis in colorectal cancer. ESMO Open. 4:e000523(3), p. 1–10. doi: 10.1136/esmoopen-2019-000523. Full text in Research Archive Show summary

  Accumulating evidence suggests immunomodulatory and context-dependent effects of TP53 mutations in cancer. We performed an exploratory analysis of the transcriptional, immunobiological and prognostic associations of TP53 mutations within the gene expression-based consensus molecular subtypes (CMSs) of colorectal cancer (CRC).Materials and methodsIn a single-hospital series of 401 stage I–IV primary CRCs, we sequenced the whole coding region of TP53 and analysed CMS-dependent transcriptional consequences of the mutations by gene expression profiling. Immunomodulatory associations were validated by multiplex, fluorescence-based immunohistochemistry of immune cell markers. Prognostic associations of TP53 mutations were analysed in an aggregated series of 635 patients classified according to CMS, including publicly available data from a French multicentre cohort (GSE39582). mutations were found in 60% of the CRCs. However, gene set enrichment analyses indicated that their transcriptional consequences varied among the CMSs and were most pronounced in CMS1-immune and CMS4-mesenchymal. Subtype specificity was primarily seen as an upregulation of gene sets reflecting cell cycle progression in CMS4 and a downregulation of T cell activity in CMS1. The subtype-dependent immunomodulatory associations were reinforced by significant depletion of several immune cell populations in mutated tumours compared with wild-type (wt) tumours exclusively in CMS1, including cytotoxic lymphocytes (adjusted p value in CMS1=0.002 and CMS2−4&gt;0.9, Microenvironment Cell Populations (MCP)-counter algorithm). This was validated by immunohistochemistry-based quantification of tumour infiltrating CD8+ cells. Within CMS1, the immunomodulatory association of TP53 mutations was strongest among microsatellite stable (MSS) tumours, and this translated into a propensity for metastatic disease and poor prognostic value of the mutations specifically in the CMS1/MSS subtype (both series overall survival: TP53 mutation vs wt: HR 5.52, p=0.028).Integration of TP53 mutation status with the CMS framework in primary CRC suggested subtype-dependent immunobiological associations with prognostic and potentially immunotherapeutic implications, warranting independent validation.

• Sveen, Anita; Kopetz, Scott & Lothe, Ragnhild A (2019). Biomarker-guided therapy for colorectal cancer: strength in complexity. Nature Reviews Clinical Oncology. ISSN 1759-4774. p. 1–22. doi: 10.1038/s41571-019-0241-1. Full text in Research Archive
• Røsok, Bård Ingvald; Brunsell, Tuva Høst; Brudvik, Kristoffer Watten; Carling, Johan Ulrik; Dorenberg, Eric Jürgen & Björnsson, B. [Show all 9 contributors for this article] (2019). Characterization of early recurrences following liver resection by ALPPS and two stage hepatectomy in patients with colorectal liver-metastases and small future liver remnants; a translational substudy of the LIGRO-RCT. HPB. ISSN 1365-182X. 21(8), p. 1017–1023. doi: 10.1016/j.hpb.2018.12.003. Full text in Research Archive
• Berg, Kaja Christine Graue; Sveen, Anita; Høland, Maren; Alagaratnam, Sharmini; Rasmussen, Marianne Berg & Danielsen, Stine Aske [Show all 9 contributors for this article] (2019). Gene expression profiles of CMS2-epithelial/canonical colorectal cancers are largely driven by DNA copy number gains. Oncogene. ISSN 0950-9232. 38(33), p. 6109–6122. doi: 10.1038/s41388-019-0868-5. Full text in Research Archive Show summary

  About 80% of colorectal cancers (CRCs) have chromosomal instability, which is an integral part of aggressive malignancy development, but the importance of specific copy number aberrations (CNAs) in modulating gene expression, particularly within the framework of clinically relevant molecular subtypes, remains mostly elusive. We performed DNA copy number profiling of 257 stage I-IV primary CRCs and integrative gene expression analysis in 151 microsatellite stable (MSS) tumors, focusing on high-level amplifications and the effect of CNAs on the characteristics of the gene expression-based consensus molecular subtypes (CMS). The results were validated in 323 MSS tumors from TCGA. Novel recurrent high-level amplifications (≥15 additional copies) with a major impact on gene expression were found for TOX3 (16q) at 1.5% frequency, as well as for CCND2 (12p) and ANXA11 (10q) at 1% frequency, in addition to the well-known targets ERBB2 (17q) and MYC (8q). Focal amplifications with ≥15 or ≥5 additional copies of at least one of these regions were associated with a poor overall survival among patients with stage I-III MSS CRCs (multivariable hazard ratio ≥3.2, p ≤ 0.01). All high-level amplifications were focal and had a more consistent relationship with gene expression than lower amplitude and/or broad-range amplifications, suggesting specific targeting during carcinogenesis. Genome-wide, copy number driven gene expression was enriched for pathways characteristic of the CMS2-epithelial/canonical subtype, including DNA repair and cell cycle progression. Furthermore, 50% of upregulated genes in CMS2-epithelial/canonical MSS CRCs were driven by CNAs, an enrichment compared with the other CMS groups, and associated with the stronger correspondence between CNAs and gene expression in malignant epithelial cells than in the cells of the tumor microenvironment (fibroblasts, endothelial cells, leukocytes). In conclusion, we identify novel recurrent amplifications with impact on gene expression in CRC and provide the first evidence that CMS2 may have a stronger copy-number related genetic basis than subtypes more heavily influenced by gene expression signals from the tumor microenvironment.

• Smeby, Jørgen; Sveen, Anita; Eilertsen, Ina Andrassy; Danielsen, Stine Aske; Hoff, Andreas Midbøe & Eide, Peter Andreas Wold [Show all 12 contributors for this article] (2019). Transcriptional and functional consequences of TP53 splice mutations in colorectal cancer. Oncogenesis. 8:35(6), p. 1–8. doi: 10.1038/s41389-019-0141-3. Full text in Research Archive Show summary

  TP53 mutations are common in colorectal cancer (CRC). Most TP53 sequencing studies have been restricted to coding regions, but recent studies have revealed that splice mutations can generate transcript variants with distinct tumorigenic and prognostic properties. Here, we performed unrestricted sequencing of all coding sequences and splice regions of TP53 in a single-hospital series of 401 primary CRCs. TP53 splice mutations were detected in 4% of the cases (N = 16), considerably more frequent than reported in major databases, and they were mutually exclusive to exon mutations. RNA sequencing revealed high-level expression of aberrant transcript variants in the majority of splice mutated tumors (75%). Most variants were predicted to produce truncated TP53 proteins, including one sample expressing the potentially oncogenic and druggable p53ψ isoform. Despite heterogeneous transcript structures, downstream transcriptional profiling revealed that TP53 splice mutations had similar effects on TP53 target gene expression and pathway activity as exonic mutations. Intriguingly, TP53 splice mutations were associated with worse 5-year relapse-free survival in stage II disease, compared to both TP53 wild-type and exon mutations (P = 0.007). These data highlight the importance of including splice regions when examining the biological and clinical consequences of TP53 mutations in CRC.

• Carm, Kristina Totland; Hoff, Andreas Midbøe; Bakken, Anne Cathrine; Axcrona, Ulrika; Axcrona, Karol & Lothe, Ragnhild A [Show all 8 contributors for this article] (2019). Interfocal heterogeneity challenges the clinical usefulness of molecular classification of primary prostate cancer. Scientific Reports. 9(1). doi: 10.1038/s41598-019-49964-7. Full text in Research Archive Show summary

  Prostate cancer is a highly heterogeneous disease and typically multiple distinct cancer foci are present at primary diagnosis. Molecular classification of prostate cancer can potentially aid the precision of diagnosis and treatment. A promising genomic classifier was published by The Cancer Genome Atlas (TCGA), successfully classifying 74% of primary prostate cancers into seven groups based on one cancer sample per patient. Here, we explore the clinical usefulness of this classification by testing the classifier's performance in a multifocal context. We analyzed 106 cancer samples from 85 distinct cancer foci within 39 patients. By somatic mutation data from whole-exome sequencing and targeted qualitative and quantitative gene expression assays, 31% of the patients were uniquely classified into one of the seven TCGA classes. Further, different samples from the same focus had conflicting classification in 12% of the foci. In conclusion, the level of both intra- and interfocal heterogeneity is extensive and must be taken into consideration in the development of clinically useful molecular classification of primary prostate cancer.

• Lopes, Nair Susana da Costa Florim Ribeiro; Bergsland, Christian Holst; Bjørnslett, Merete Pauline; Pellinen, Teijo Severi; Svindland, Aud & Nesbakken, Arild [Show all 10 contributors for this article] (2019). Digital image analysis of multiplex fluorescence IHC in colorectal cancer recognizes the prognostic value of CDX2 and its negative correlation with SOX2. Laboratory Investigation. ISSN 0023-6837. 100(1), p. 120–134. doi: 10.1038/s41374-019-0336-4. Full text in Research Archive Show summary

  Flourescence-based multiplex immunohistochemistry (mIHC) combined with multispectral imaging and digital image analysis (DIA) is a quantitative high-resolution method for determination of protein expression in tissue. We applied this method for five biomarkers (CDX2, SOX2, SOX9, E-cadherin, and β-catenin) using tissue microarrays of a Norwegian unselected series of primary colorectal cancer. The data were compared with previously obtained chromogenic IHC data of the same tissue cores, visually assessed by the Allred method. We found comparable results between the methods, although confirmed that DIA offered improved resolution to differentiate cases with high and low protein expression. However, we experienced inherent challenges with digital image analysis of membrane staining, which was better assessed visually. DIA and mIHC enabled quantitative analysis of biomarker coexpression on the same tissue section at the single-cell level, revealing a strong negative correlation between the differentiation markers CDX2 and SOX2. Both methods confirmed known prognostic associations for CDX2, but DIA improved data visualization and detection of clinicopathological and biological associations. In summary, mIHC combined with DIA is an efficient and reliable method to evaluate protein expression in tissue, here shown to recapitulate and improve detection of known clinicopathological and survival associations for the emerging biomarker CDX2, and is therefore a candidate approach to standardize CDX2 detection in pathology laboratories.

• Burocziova, Monika; Burdova, Kamila; Martinikova, Andra S.; Kasparek, Petr; Kleiblova, Petra & Danielsen, Stine Aske [Show all 24 contributors for this article] (2019). Truncated PPM1D impairs stem cell response to genotoxic stress and promotes growth of APC-deficient tumors in the mouse colon. Cell Death & Disease. 10:818(11), p. 1–15. doi: 10.1038/s41419-019-2057-4. Full text in Research Archive
• Eide, Peter Andreas Wold; Eilertsen, Ina Andrassy; Sveen, Anita & Lothe, Ragnhild A (2019). Long noncoding RNA MIR31HG is a bona fide prognostic marker with colorectal cancer cell-intrinsic properties. International Journal of Cancer. ISSN 0020-7136. p. 1–12. doi: 10.1002/ijc.31998. Full text in Research Archive Show summary

  Elevated miR‐31 expression is associated with poor outcome in colorectal cancer (CRC). Whether the prognostic information is independent of known molecular subgroups and gene expression‐based consensus molecular subtypes (CMS) is currently unknown. To investigate this, we analyzed nearly 2000 CRC biopsies and preclinical models. The expression of miR‐31‐5p and its host transcript, long noncoding RNA MIR31HG, was strongly correlated (Spearman's ρ > 0.80). MIR31HG outlier expression was observed in 158/1265 (12%) of pCRCs and was associated with depletion of CMS2‐canonical subgroup (odds ratio = 0.21 [0.11–0.35]) and shorter relapse‐free survival (RFS) in multivariable analysis (adjusted hazard ratio = 2.2 [1.6–3.0]). For stage II disease, 5‐year RFS for patients with MIR31HG outlier status was 49% compared to 77% for those with normal‐like expression. MIR31HG outlier status was associated with inferior outcome also within clinical high risk groups and within the poor prognostic CMS4‐mesenchymal gene expression subtype specifically. Preclinical models with MIR31HG outlier expression were characterized by reduced expression of MYC targets as well as elevated epithelial‐mesenchymal transition, TNF‐α/NFκB, TGF‐β, and IFN‐α/γ gene expression signatures, indicating cancer cell‐intrinsic properties resembling the CMS4 subgroup—associations which were recapitulated in patient biopsies. Moreover, the prognostic value of MIR31HG outlier status was independent of cytotoxic T lymphocyte and fibroblast infiltration. We here present evidence that MIR31HG expression provides clinical stratification beyond major gene expression phenotypes and tumor immune and stromal cell infiltration and propose a model where increased expression is an indicator of a cellular state conferring intrinsic invasive and/or immuno‐evasive capabilities.

• Kryeziu, Kushtrim; Bruun, Jarle; Guren, Tormod Kyrre; Sveen, Anita & Lothe, Ragnhild A (2019). Combination therapies with HSP90 inhibitors against colorectal cancer. Biochimica et Biophysica Acta. CR. Reviews on Cancer. ISSN 0304-419X. 1871(2), p. 240–247. doi: 10.1016/j.bbcan.2019.01.002. Full text in Research Archive Show summary

  Oncogene stability and homeostasis mediated by the HSP90 chaperone is a crucial protection trait of cancer cells. Therefore, HSP90 represents an attractive therapeutic target for many cancers, including colorectal cancer. Although monotherapy has limited clinical efficacy, preclinical and early-phase clinical studies indicate improved antitumor activity when HSP90 inhibitors are combined with chemotherapies or targeted agents. This may be further improved with a biomarker-guided approach based on oncogenic HSP90 clients, or stratification based on the consensus molecular subtypes of colorectal cancer, suggesting a synergistic activity with 5-fluorouracil in preclinical models of the chemorefractory mesenchymal subtype. Furthermore, HSP90 inhibition may activate mechanisms to turn non-immunogenic tumors hot and improve their recognition by the immune system, suggesting synergy with immune checkpoint blockade.

• Brunsell, Tuva Høst; Cengija, Vanja; Sveen, Anita; Bjørnbeth, Bjørn Atle; Røsok, Bård Ingvald & Brudvik, Kristoffer Watten [Show all 10 contributors for this article] (2019). Heterogeneous radiological response to neoadjuvant therapy is associated with poor prognosis after resection of colorectal liver metastases. European Journal of Surgical Oncology. ISSN 0748-7983. p. 1–7. doi: 10.1016/j.ejso.2019.07.017. Full text in Research Archive Show summary

  Introduction Surgery combined with perioperative chemotherapy has become standard of care in patients with resectable colorectal liver metastases. However, poor outcome is expected for a significant subgroup. The clinical implications of inter-metastatic heterogeneity remain largely unknown. In a prospective, population-based series of patients undergoing resection of multiple colorectal liver metastases, the aim was to investigate the prevalence and prognostic impact of heterogeneous response to neoadjuvant chemotherapy. Materials and Methods Radiological response to treatment was evaluated in a lesion-specific manner in 2–5 metastases per patient. Change of lesion diameter was evaluated and response/progression was classified according to three different size thresholds; 3, 4 and 5 mm. A heterogeneous response was defined as progression and response of different metastases in the same patient. Results In total, 142 patients with 585 liver metastases were examined with the same radiological method (MRI or CT) before and after neoadjuvant treatment. Heterogeneous response to treatment was seen in 16 patients (11%) using the 3 mm size change threshold, and this group had a 5-year cancer-specific survival of 19% compared to 49% for patients with response in all lesions (p = 0.003). Cut-off values of 4–5 mm were less sensitive for detecting a heterogeneous response, but the survival difference was similar and significant. Conclusion A subgroup of patients with multiple colorectal liver metastases had heterogeneous radiological response to neoadjuvant chemotherapy and poor prognosis. The evaluation of response pattern is easy to perform, feasible in clinical practice and, if validated, a promising biomarker for treatment decisions.

• Dienstmann, Rodrigo; Villacampa, Guillermo; Sveen, Anita; Mason, Michael J.; Niedzwiecki, Donna & Nesbakken, Arild [Show all 10 contributors for this article] (2019). Relative contribution of clinicopathological variables, genomic markers, transcriptomic subtyping and microenvironment features for outcome prediction in stage II/III colorectal cancer. Annals of Oncology. ISSN 0923-7534. 30(10), p. 1622–1629. doi: 10.1093/annonc/mdz287. Full text in Research Archive Show summary

  Background It remains unknown to what extent consensus molecular subtype (CMS) groups and immune-stromal infiltration patterns improve our ability to predict outcomes over tumor–node–metastasis (TNM) staging and microsatellite instability (MSI) status in early-stage colorectal cancer (CRC). Patients and methods We carried out a comprehensive retrospective biomarker analysis of prognostic markers in adjuvant chemotherapy-untreated (N = 1656) and treated (N = 980), stage II (N = 1799) and III (N = 837) CRCs. We defined CMS scores and estimated CD8+ cytotoxic lymphocytes (CytoLym) and cancer-associated fibroblasts (CAF) infiltration scores from bulk tumor tissue transcriptomes (CMSclassifier and MCPcounter R packages); constructed a stratified multivariable Cox model for disease-free survival (DFS); and calculated the relative proportion of explained variation by each marker (clinicopathological [ClinPath], genomics [Gen: MSI, BRAF and KRAS mutations], CMS scores [CMS] and microenvironment cells [MicroCells: CytoLym+CAF]). Results In multivariable models, only ClinPath and MicroCells remained significant prognostic factors, with both CytoLym and CAF infiltration scores improving survival prediction beyond other markers. The explained variation for DFS models of ClinPath, MicroCells, Gen markers and CMS4 scores was 77%, 14%, 5.3% and 3.7%, respectively, in stage II; and 55.9%, 35.1%, 4.1% and 0.9%, respectively, in stage III. Patients whose tumors were CytoLym high/CAF low had better DFS than other strata [HR=0.71 (0.6–0.9); P = 0.004]. Microsatellite stable tumors had the strongest signal for improved outcomes with CytoLym high scores (interaction P = 0.04) and the poor prognosis linked to high CAF scores was limited to stage III disease (interaction P = 0.04). Conclusions Our results confirm that tumor microenvironment infiltration patterns represent potent determinants of the risk for distant dissemination in early-stage CRC. Multivariable models suggest that the prognostic value of MSI and CMS groups is largely explained by CytoLym and CAF infiltration patterns.

• Pinto, Diana; Pinto, Carla; Guerra, Joana; Pinheiro, Manuela; Santos, Rui & Vedeld, Hege Marie [Show all 15 contributors for this article] (2018). Contribution of MLH1 constitutional methylation for Lynch syndrome diagnosis in patients with tumor MLH1 downregulation. Cancer Medicine. 7(2), p. 433–444. doi: 10.1002/cam4.1285. Full text in Research Archive Show summary

  Constitutional epimutation of the two major mismatch repair genes, MLH1 and MSH2, has been identified as an alternative mechanism that predisposes to the development of Lynch syndrome. In the present work, we aimed to investigate the prevalence of MLH1 constitutional methylation in colorectal cancer (CRC) patients with abnormal expression of the MLH1 protein in their tumors. In a series of 38 patients who met clinical criteria for Lynch syndrome genetic testing, with loss of MLH1 expression in the tumor and with no germline mutations in the MLH1 gene (35/38) or with tumors presenting the BRAF p.Val600Glu mutation (3/38), we screened for constitutional methylation of the MLH1 gene promoter using methylation‐specific multiplex ligation‐dependent probe amplification (MS‐MLPA) in various biological samples. We found four (4/38; 10.5%) patients with constitutional methylation in the MLH1 gene promoter. RNA studies demonstrated decreased MLH1 expression in the cases with constitutional methylation when compared with controls. We could infer the mosaic nature of MLH1 constitutional hypermethylation in tissues originated from different embryonic germ layers, and in one family we could show that it occurred de novo. We conclude that constitutional MLH1 methylation occurs in a significant proportion of patients who have loss of MLH1 protein expression in their tumors and no MLH1 pathogenic germline mutation. Furthermore, we provide evidence that MLH1 constitutional hypermethylation is the molecular mechanism behind about 3% of Lynch syndrome families diagnosed in our institution, especially in patients with early onset or multiple primary tumors without significant family history.

• Eilertsen, Ina Andrassy; Sveen, Anita; Strømme, Jonas Meier; Skotheim, Rolf I.; Nesbakken, Arild & Lothe, Ragnhild A (2018). Alternative splicing expands the prognostic impact of KRAS in microsatellite stable primary colorectal cancer. International Journal of Cancer. ISSN 0020-7136. 144(4), p. 841–847. doi: 10.1002/ijc.31809. Full text in Research Archive Show summary

  KRAS mutation is a well‐known marker for poor response to targeted treatment and patient prognosis in microsatellite stable (MSS) colorectal cancer (CRC). However, variation in clinical outcomes among patients wild‐type for KRAS underlines that this is not a homogeneous population. Here, we evaluated the prognostic impact of KRAS alternative splicing in relation to mutation status in a single‐hospital series of primary MSS CRCs (N = 258). Using splicing‐sensitive microarrays and RNA sequencing, the relative expression of KRAS‐4A versus KRAS‐4B transcript variants was confirmed to be down‐regulated in CRC compared to normal colonic mucosa (N = 41; p ≤ 0.001). This was independent of mutation status, however, gene set enrichment analysis revealed that the effect of splicing on KRAS signaling was specific to the KRAS wild‐type subgroup, in which low relative KRAS‐4A expression was associated with a higher level of KRAS signaling (p = 0.005). In concordance, the prognostic value of KRAS splicing was also dependent on mutation status, and for patients with Stage I–III KRAS wild‐type MSS CRC, low relative KRAS‐4A expression was associated with inferior overall survival (HR: 2.36, 95% CI: 1.07–5.18, p = 0.033), a result not found in mutant cases (pinteraction = 0.026). The prognostic association in the wild‐type subgroup was independent of clinicopathological factors, including cancer stage in multivariable analysis (HR: 2.68, 95% CI: 1.18–6.09, p = 0.018). This suggests that KRAS has prognostic value beyond mutation status in MSS CRC, and highlights the importance of molecular heterogeneity in the clinically relevant KRAS wild‐type subgroup.

• Høland, Maren; Kolberg, Matthias; Danielsen, Stine Aske; Bjerkehagen, Bodil; Eilertsen, Ina Andrassy & Hektoen, Merete [Show all 14 contributors for this article] (2018). Inferior survival for patients with malignant peripheral nerve sheath tumors defined by aberrant TP53. Modern Pathology. ISSN 0893-3952. p. 1–14. doi: 10.1038/s41379-018-0074-y. Full text in Research Archive Show summary

  Malignant peripheral nerve sheath tumor is a rare and aggressive disease with poor treatment response, mainly affecting adolescents and young adults. Few molecular biomarkers are used in the management of this cancer type, and although TP53 is one of few recurrently mutated genes in malignant peripheral nerve sheath tumor, the mutation prevalence and the corresponding clinical value of the TP53 network remains unsettled. We present a multi-level molecular study focused on aberrations in the TP53 network in relation to patient outcome in a series of malignant peripheral nerve sheath tumors from 100 patients and 38 neurofibromas, including TP53 sequencing, high-resolution copy number analyses of TP53 and MDM2, and gene expression profiling. Point mutations in TP53 were accompanied by loss of heterozygosity, resulting in complete loss of protein function in 8.2% of the malignant peripheral nerve sheath tumors. Another 5.5% had MDM2 amplification. TP53 mutation and MDM2 amplification were mutually exclusive and patients with either type of aberration in their tumor had a worse prognosis, compared to those without (hazard ratio for 5-year disease-specific survival 3.5, 95% confidence interval 1.78–6.98). Both aberrations had similar consequences on the gene expression level, as analyzed by a TP53-associated gene signature, a property also shared with the copy number aberrations and/or loss of heterozygosity at the TP53 locus, suggesting a common “TP53-mutated phenotype” in as many as 60% of the tumors. This was a poor prognostic phenotype (hazard ratio = 4.1, confidence interval:1.7–9.8), thus revealing a TP53-non-aberrant patient subgroup with a favorable outcome. The frequency of the “TP53-mutated phenotype” warrants explorative studies of stratified treatment strategies in malignant peripheral nerve sheath tumor.

• Danielsen, Håvard Emil; Hveem, Tarjei Sveinsgjerd; Domingo, Enric; Pradhan, Manohar; Kleppe, Andreas & Syvertsen, Rolf Anders [Show all 18 contributors for this article] (2018). Prognostic markers for colorectal cancer: Estimating ploidy and stroma. Annals of Oncology. ISSN 0923-7534. 29(3), p. 616–623. doi: 10.1093/annonc/mdx794. Full text in Research Archive
• Oltedal, Satu; Kørner, Hartwig; Aasprong, Ole Gunnar; Hussain, Israr; Tjensvoll, Kjersti & Smaaland, Rune [Show all 12 contributors for this article] (2018). The Prognostic Relevance of Sentinel Lymph Node Metastases Assessed by PHGR1 mRNA Quantification in Stage I to III Colon Cancer. Translational Oncology. ISSN 1944-7124. 11(2), p. 436–443. doi: 10.1016/j.tranon.2018.01.015. Full text in Research Archive
• Pharo, Heidi Dietrichson; Andresen, Kim; Berg, Kaja Christine Graue; Lothe, Ragnhild A; Jeanmougin, Marine & Lind, Guro Elisabeth (2018). A robust internal control for high-precision DNA methylation analyses by droplet digital PCR. Clinical Epigenetics. 10(24). doi: 10.1186/s13148-018-0456-5. Full text in Research Archive Show summary

  Background Droplet digital PCR (ddPCR) allows absolute quantification of nucleic acids and has potential for improved non-invasive detection of DNA methylation. For increased precision of the methylation analysis, we aimed to develop a robust internal control for use in methylation-specific ddPCR. Methods Two control design approaches were tested: (a) targeting a genomic region shared across members of a gene family and (b) combining multiple assays targeting different pericentromeric loci on different chromosomes. Through analyses of 34 colorectal cancer cell lines, the performance of the control assay candidates was optimized and evaluated, both individually and in various combinations, using the QX200™ droplet digital PCR platform (Bio-Rad). The best-performing control was tested in combination with assays targeting methylated CDO1, SEPT9, and VIM. Results A 4Plex panel consisting of EPHA3, KBTBD4, PLEKHF1, and SYT10 was identified as the best-performing control. The use of the 4Plex for normalization reduced the variability in methylation values, corrected for differences in template amount, and diminished the effect of chromosomal aberrations. Positive Droplet Calling (PoDCall), an R-based algorithm for standardized threshold determination, was developed, ensuring consistency of the ddPCR results. Conclusion Implementation of a robust internal control, i.e., the 4Plex, and an algorithm for automated threshold determination, PoDCall, in methylation-specific ddPCR increase the precision of DNA methylation analysis.

• Smeby, Jørgen; Sveen, Anita; Merok, Marianne Aarstad; Danielsen, Stine Aske; Eilertsen, Ina Andrassy & Guren, Marianne [Show all 9 contributors for this article] (2018). CMS-dependent prognostic impact of KRAS and BRAFV600E mutations in primary colorectal cancer. Annals of Oncology. ISSN 0923-7534. 29(5), p. 1227–1234. doi: 10.1093/annonc/mdy085. Full text in Research Archive Show summary

  Background: The prognostic impact of KRAS and BRAFV600E mutations in primary colorectal cancer (CRC) varies with microsatellite instability (MSI) status. The gene expression–based consensus molecular subtypes (CMSs) of CRC define molecularly and clinically distinct subgroups, and represent a novel stratification framework in biomarker analysis. We investigated the prognostic value of these mutations within the CMS groups. Patients and methods: Totally 1197 primary tumors from a Norwegian series of CRC stage I–IV were analyzed for MSI and mutation status in hotspots in KRAS (codons 12, 13 and 61) and BRAF (codon 600). A subset was analyzed for gene expression and confident CMS classification was obtained for 317 samples. This cohort was expanded with clinical and molecular data, including CMS classification, from 514 patients in the publically available dataset GSE39582. Gene expression signatures associated with KRAS and BRAFV600E mutations were used to evaluate differential impact of mutations on gene expression among the CMS groups. Results: BRAFV600E and KRAS mutations were both associated with inferior 5-year overall survival (OS) exclusively in MSS tumors (BRAFV600E mutation versus KRAS/BRAF wild-type: Hazard ratio (HR) 2.85, P < 0.001; KRAS mutation versus KRAS/BRAF wild-type: HR 1.30, P = 0.013). BRAFV600E-mutated MSS tumors were strongly enriched and associated with metastatic disease in CMS1, leading to negative prognostic impact in this subtype (OS: BRAFV600E mutation versus wild-type: HR 7.73, P = 0.001). In contrast, the poor prognosis of KRAS mutations was limited to MSS tumors with CMS2/CMS3 epithelial-like gene expression profiles (OS: KRAS mutation versus wild-type: HR 1.51, P = 0.011). The subtype-specific prognostic associations were substantiated by differential effects of BRAFV600E and KRAS mutations on gene expression signatures according to the MSI status and CMS group. Conclusions: BRAFV600E mutations are enriched and associated with metastatic disease in CMS1 MSS tumors, leading to poor prognosis in this subtype. KRAS mutations are associated with adverse outcome in epithelial (CMS2/CMS3) MSS tumors.

• Bruun, Jarle; Sveen, Anita; Barros, Rita; Eide, Peter Andreas Wold; Eilertsen, Ina Andrassy & Kolberg, Matthias [Show all 14 contributors for this article] (2018). Prognostic, predictive, and pharmacogenomic assessments of CDX2 refine stratification of colorectal cancer. Molecular Oncology. ISSN 1574-7891. 12(9), p. 1639–1655. doi: 10.1002/1878-0261.12347. Full text in Research Archive Show summary

  We aimed to refine the value of CDX2 as an independent prognostic and predictive biomarker in colorectal cancer (CRC) according to disease stage and chemotherapy sensitivity in preclinical models. CDX2 expression was evaluated in 1045 stage I–IV primary CRCs by gene expression (n = 403) or immunohistochemistry (n = 642) and in relation to 5‐year relapse‐free survival (RFS), overall survival (OS), and chemotherapy. Pharmacogenomic associations between CDX2 expression and 69 chemotherapeutics were assessed by drug screening of 35 CRC cell lines. CDX2 expression was lost in 11.6% of cases and showed independent poor prognostic value in multivariable models. For individual stages, CDX2 was prognostic only in stage IV, independent of chemotherapy. Among stage I–III patients not treated in an adjuvant setting, CDX2 loss was associated with a particularly poor survival in the BRAF‐mutated subgroup, but prognostic value was independent of microsatellite instability status and the consensus molecular subtypes. In stage III, the 5‐year RFS rate was higher among patients with loss of CDX2 who received adjuvant chemotherapy than among patients who did not. The CDX2‐negative cell lines were significantly more sensitive to chemotherapeutics than CDX2‐positive cells, and the multidrug resistance genes MDR1 and CFTR were significantly downregulated both in CDX2‐negative cells and in patient tumors. Loss of CDX2 in CRC is an adverse prognostic biomarker only in stage IV disease and appears to be associated with benefit from adjuvant chemotherapy in stage III. Early‐stage patients not qualifying for chemotherapy might be reconsidered for such treatment if their tumor has loss of CDX2 and mutated BRAF.

• Løvf, Marthe; Zhao, Sen; Axcrona, Ulrika; Johannessen, Bjarne; Bakken, Anne Cathrine & Carm, Kristina Totland [Show all 13 contributors for this article] (2018). Multifocal primary prostate cancer exhibits high degree of genomic heterogeneity. European Urology. ISSN 0302-2838. p. 1–8. doi: 10.1016/j.eururo.2018.08.009. Full text in Research Archive
• Vedeld, Hege Marie; Nesbakken, Arild; Lothe, Ragnhild A & Lind, Guro Elisabeth (2018). Re-assessing ZNF331 as a DNA methylation biomarker for colorectal cancer. Clinical Epigenetics. 10(1). doi: 10.1186/s13148-018-0503-2. Full text in Research Archive Show summary

  We have previously shown that aberrant promoter methylation of ZNF331 is a potential biomarker for colorectal cancer detection with high sensitivity (71%) and specificity (98%). This finding was recently confirmed by others, and it was additionally suggested that promoter methylation of ZNF331 was an independent prognostic biomarker for colorectal cancer (n = 146). In the current study, our initial colorectal cancer sample series was extended to include a total of 423 cancer tissue samples. Aberrant promoter methylation was found in 71% of the samples, thus repeatedly suggesting the biomarker potential of ZNF331 for detection of colorectal cancer. Furthermore, multivariate Cox’s analysis indicated a trend towards inferior overall survival for colorectal cancer patients with aberrant methylation of ZNF331.

• Berg, Kaja Christine Graue; Eide, Peter Andreas Wold; Eilertsen, Ina Andrassy; Johannessen, Bjarne; Bruun, Jarle & Danielsen, Stine Aske [Show all 14 contributors for this article] (2017). Multi-omics of 34 colorectal cancer cell lines - a resource for biomedical studies. Molecular Cancer. 16:116, p. 1–16. doi: 10.1186/s12943-017-0691-y. Full text in Research Archive
• Sveen, Anita; Bruun, Jarle; Eide, Peter Andreas Wold; Eilertsen, Ina Andrassy; Ramirez, Lorena & Murumagi, Astrid [Show all 17 contributors for this article] (2017). Colorectal cancer consensus molecular subtypes translated to preclinical models uncover potentially targetable cancer cell dependencies. Clinical Cancer Research. ISSN 1078-0432. 24(4), p. 794–806. doi: 10.1158/1078-0432.CCR-17-1234. Full text in Research Archive
• Sveen, Anita; Johannessen, Bjarne; Tengs, Torstein; Danielsen, Stine Aske; Eilertsen, Ina Andrassy & Lind, Guro Elisabeth [Show all 16 contributors for this article] (2017). Multilevel genomics of colorectal cancers with microsatellite instability – clinical impact of JAK1 mutations and consensus molecular subtype 1. Genome Medicine. 9(1). doi: 10.1186/s13073-017-0434-0. Full text in Research Archive Show summary

  Background Approximately 15% of primary colorectal cancers have DNA mismatch repair deficiency, causing a complex genome with thousands of small mutations—the microsatellite instability (MSI) phenotype. We investigated molecular heterogeneity and tumor immunogenicity in relation to clinical endpoints within this distinct subtype of colorectal cancers. Methods A total of 333 primary MSI+ colorectal tumors from multiple cohorts were analyzed by multilevel genomics and computational modeling—including mutation profiling, clonality modeling, and neoantigen prediction in a subset of the tumors, as well as gene expression profiling for consensus molecular subtypes (CMS) and immune cell infiltration. Results Novel, frequent frameshift mutations in four cancer-critical genes were identified by deep exome sequencing, including in CRTC1, BCL9, JAK1, and PTCH1. JAK1 loss-of-function mutations were validated with an overall frequency of 20% in Norwegian and British patients, and mutated tumors had up-regulation of transcriptional signatures associated with resistance to anti-PD-1 treatment. Clonality analyses revealed a high level of intra-tumor heterogeneity; however, this was not associated with disease progression. Among the MSI+ tumors, the total mutation load correlated with the number of predicted neoantigens (P = 4 × 10−5), but not with immune cell infiltration—this was dependent on the CMS class; MSI+ tumors in CMS1 were highly immunogenic compared to MSI+ tumors in CMS2-4. Both JAK1 mutations and CMS1 were favorable prognostic factors (hazard ratios 0.2 [0.05–0.9] and 0.4 [0.2–0.9], respectively, P = 0.03 and 0.02). Conclusions Multilevel genomic analyses of MSI+ colorectal cancer revealed molecular heterogeneity with clinical relevance, including tumor immunogenicity and a favorable patient outcome associated with JAK1 mutations and the transcriptomic subgroup CMS1, emphasizing the potential for prognostic stratification of this clinically important subtype. See related research highlight by Samstein and Chan 10.1186/s13073-017-0438-9

• Dienstmann, Rodrigo; Mason, M; Sinicrope, FA; Phipps, Alison; Tejpar, S. & Nesbakken, Arild [Show all 14 contributors for this article] (2017). Prediction of overall survival in stage II and III colon cancer beyond TNM system: a retrospective, pooled biomarker study. Annals of Oncology. ISSN 0923-7534. 28(5), p. 1023–1031. doi: 10.1093/annonc/mdx052. Full text in Research Archive Show summary

  Background: TNM staging alone does not accurately predict outcome in colon cancer (CC) patients who may be eligible for adjuvant chemotherapy. It is unknown to what extent the molecular markers microsatellite instability (MSI) and mutations in BRAF or KRAS improve prognostic estimation in multivariable models that include detailed clinicopathological annotation. Patients and methods: After imputation of missing at random data, a subset of patients accrued in phase 3 trials with adjuvant chemotherapy (n = 3016)—N0147 (NCT00079274) and PETACC3 (NCT00026273)—was aggregated to construct multivariable Cox models for 5-year overall survival that were subsequently validated internally in the remaining clinical trial samples (n = 1499), and also externally in different population cohorts of chemotherapy-treated (n = 949) or -untreated (n = 1080) CC patients, and an additional series without treatment annotation (n = 782). Results: TNM staging, MSI and BRAFV600E mutation status remained independent prognostic factors in multivariable models across clinical trials cohorts and observational studies. Concordance indices increased from 0.61–0.68 in the TNM alone model to 0.63–0.71 in models with added molecular markers, 0.65–0.73 with clinicopathological features and 0.66–0.74 with all covariates. In validation cohorts with complete annotation, the integrated time-dependent AUC rose from 0.64 for the TNM alone model to 0.67 for models that included clinicopathological features, with or without molecular markers. In patient cohorts that received adjuvant chemotherapy, the relative proportion of variance explained (R2) by TNM, clinicopathological features and molecular markers was on an average 65%, 25% and 10%, respectively. Conclusions: Incorporation of MSI, BRAFV600E and KRAS mutation status to overall survival models with TNM staging improves the ability to precisely prognosticate in stage II and III CC patients, but only modestly increases prediction accuracy in multivariable models that include clinicopathological features, particularly in chemotherapy-treated patients.

• Landskron, Johannes; Kraggerud, Sigrid M.; Wik, Elisabeth; Dørum, Anne; Bjørnslett, Merete Pauline & Melum, Espen [Show all 11 contributors for this article] (2017). C77G in PTPRC (CD45) is no risk allele for ovarian cancer, but associated with less aggressive disease. PLOS ONE. 12:e0182030(7), p. 1–12. doi: 10.1371/journal.pone.0182030. Full text in Research Archive Show summary

  The pan lymphocyte marker CD45 exists in various isoforms arising from alternative splicing of the exons 4, 5 and 6. While naïve T cells express CD45RA translated from an mRNA containing exon 4, exons 4–6 are spliced out to encode the shorter CD45R0 in antigen-experienced effector/memory T cells. The SNP C77G (rs17612648) is located in exon 4 and blocks the exon’s differential splicing from the pre-mRNA, enforcing expression of CD45RA. Several studies have linked C77G to autoimmune diseases but lack of validation in other cohorts has left its role elusive. An incidental finding in an ovarian cancer patient cohort from West Norway (Bergen region, n = 312), suggested that the frequency of C77G was higher among ovarian cancer patients than in healthy Norwegians (n = 1,357) (3.0% vs. 1.8% allele frequency). However, this finding could not be validated in a larger patient cohort from South-East Norway (Oslo region, n = 1,198) with 1.2% allele frequency. Hence, C77G is not associated with ovarian cancer in the Norwegian population. However, its frequency was increased in patients with FIGO stage II, endometrioid histology or an age at diagnosis of 60 years or older indicating a possible association with a less aggressive cancer type.

• Sveen, Anita; Bruun, Jarle; Eide, Peter W.; Eilertsen, Ina A.; Ramirez, Lorena & Murumagi, Astrid [Show all 17 contributors for this article] (2017). Colorectal cancer consensus molecular subtypes define selective activity of HSP90 inhibitors in preclinical models. Cancer Reports and Reviews. doi: 10.1158/1078-0432.CCR-17-1234. Full text in Research Archive
• Lind, Guro Elisabeth; Guriby, Marianne; Ahlquist, Terje Cruickshank; Hussain, Israr; Jeanmougin, Marine & Søreide, Kjetil [Show all 9 contributors for this article] (2017). Prognostic relevance of an epigenetic biomarker panel in sentinel lymph nodes from colon cancer patients. Clinical Epigenetics. 9:97, p. 1–8. doi: 10.1186/s13148-017-0397-4. Full text in Research Archive Show summary

  Background: Patients with early colorectal cancer (stages I–II) generally have a good prognosis, but a subgroup of 15–20% experiences relapse and eventually die of disease. Occult metastases have been suggested as a marker for increased risk of recurrence in patients with node-negative disease. Using a previously identified, highly accurate epigenetic biomarker panel for early detection of colorectal tumors, we aimed at evaluating the prognostic value of occult metastases in sentinel lymph nodes of colon cancer patients. Results: The biomarker panel was analyzed by quantitative methylation-specific PCR in primary tumors and 783 sentinel lymph nodes from 201 patients. The panel status in sentinel lymph nodes showed a strong association with lymph node stage (P = 8.2E−17). Compared with routine lymph node diagnostics, the biomarker panel had a sensitivity of 79% (31/39). Interestingly, among 162 patients with negative lymph nodes from routine diagnostics, 13 (8%) were positive for the biomarker panel. Colon cancer patients with high sentinel lymph node methylation had an inferior prognosis (5-year overall survival P = 3.0E−4; time to recurrence P = 3.1E−4), although not significant. The same trend was observed in multivariate analyses (P = 1.4E−1 and P = 6.7E−2, respectively). Occult sentinel lymph node metastases were not detected in early stage (I–II) colon cancer patients who experienced relapse. Conclusions: Colon cancer patients with high sentinel lymph node methylation of the analyzed epigenetic biomarker panel had an inferior prognosis, although not significant in multivariate analyses. Occult metastases in TNM stage II patients that experienced relapse were not detected.

• Totland, Max; Bergsland, Christian Holst; Fykerud, Tone Aase; Knudsen, Lars Mørland; Rasmussen, Nikoline L. & Eide, Peter Andreas Wold [Show all 11 contributors for this article] (2017). The E3 ubiquitin ligase NEDD4 induces endocytosis and lysosomal sorting of connexin 43 to promote loss of gap junctions. Journal of Cell Science. ISSN 0021-9533. 130(17), p. 2867–2882. doi: 10.1242/jcs.202408. Full text in Research Archive
• Kolberg, Matthias; Bruun, Jarle; Murumagi, Astrid; Mpindi, John P.; Bergsland, Christian Holst & Høland, Maren [Show all 10 contributors for this article] (2017). Drug sensitivity and resistance testing identifies PLK1 inhibitors and gemcitabine as potent drugs for malignant peripheral nerve sheath tumors. Molecular Oncology. ISSN 1574-7891. 11(9), p. 1156–1171. doi: 10.1002/1878-0261.12086. Full text in Research Archive
• Ramalho-Carvalho, João; Martins, João Barbosa; Cekaite, Lina; Sveen, Anita; Torres-Ferreira, Jorge & Graça, Inês [Show all 13 contributors for this article] (2017). Epigenetic disruption of miR-130a promotes prostate cancer by targeting SEC23B and DEPDC1. Cancer Letters. ISSN 0304-3835. 385, p. 150–159. doi: 10.1016/j.canlet.2016.10.028. Full text in Research Archive
• Vedeld, Hege Marie; Merok, Marianne Aarstad; Jeanmougin, Marine; Danielsen, Stine Aske; Honne, Hilde & Presthus, Gro [Show all 13 contributors for this article] (2017). CpG island methylator phenotype identifies high risk patients among microsatellite stable BRAF mutated colorectal cancers. International Journal of Cancer. ISSN 0020-7136. 141(5), p. 967–976. doi: 10.1002/ijc.30796. Full text in Research Archive
• Eide, Peter Andreas Wold; Bruun, Jarle; Lothe, Ragnhild A & Sveen, Anita (2017). CMScaller: An R package for consensus molecular subtyping of colorectal cancer pre-clinical models. Scientific Reports. 7(1). doi: 10.1038/s41598-017-16747-x. Full text in Research Archive Show summary

  Colorectal cancers (CRCs) can be divided into four gene expression-based biologically distinct consensus molecular subtypes (CMS). This classification provides a potential framework for stratified treatment, but to identify novel CMS-drug associations, translation of the subtypes to pre-clinical models is essential. The currently available classifier is dependent on gene expression signals from the immune and stromal compartments of tumors and fails to identify the poor-prognostic CMS4-mesenchymal group in immortalized cell lines, patient-derived organoids and xenografts. To address this, we present a novel CMS classifier based on a filtered set of cancer cell-intrinsic, subtype-enriched gene expression markers. This new classifier, referred to as CMScaller, recapitulated the subtypes in both in vitro and in vivo models (551 in total). Importantly, by analyzing public drug response data from patient-derived xenografts and cell lines, we show that the subtypes are predictive of response to standard CRC drugs. CMScaller is available as an R package.

• Sveen, Anita; Kilpinen, Sami; Ruusulehto, Anja; Lothe, Ragnhild A & Skotheim, Rolf Inge (2016). Aberrant RNA splicing in cancer; Expression changes and driver mutations of splicing factor genes. Oncogene. ISSN 0950-9232. 35(19), p. 2413–2427. doi: 10.1038/onc.2015.318. Full text in Research Archive
• Hoff, Andreas Midbøe; Alagaratnam, Sharmini; Zhao, Sen; Bruun, Jarle; Andrews, PW & Lothe, Ragnhild A [Show all 7 contributors for this article] (2016). Identification of novel fusion genes in testicular germ cell tumors. Cancer Research. ISSN 0008-5472. 76(1), p. 108–116. doi: 10.1158/0008-5472.CAN-15-1790. Full text in Research Archive
• Lorenz, Susanne; Barøy, Tale; Sun, Jinchang; Nome, Torfinn; Vodak, Daniel & Bryne, Jan Christian [Show all 21 contributors for this article] (2016). Unscrambling the genomic chaos of osteosarcoma reveals extensive transcript fusion, recurrent rearrangements and frequent novel TP53 aberrations. OncoTarget. 7(5), p. 5273–5288. doi: 10.18632/oncotarget.6567. Full text in Research Archive Show summary

  In contrast to many other sarcoma subtypes, the chaotic karyotypes of osteosarcoma have precluded the identification of pathognomonic translocations. We here report hundreds of genomic rearrangements in osteosarcoma cell lines, showing clear characteristics of microhomology-mediated break-induced replication (MMBIR) and end-joining repair (MMEJ) mechanisms. However, at RNA level, the majority of the fused transcripts did not correspond to genomic rearrangements, suggesting the involvement of trans-splicing, which was further supported by typical trans-splicing characteristics. By combining genomic and transcriptomic analysis, certain recurrent rearrangements were identified and further validated in patient biopsies, including a PMP22-ELOVL5 gene fusion, genomic structural variations affecting RB1, MTAP/CDKN2A and MDM2, and, most frequently, rearrangements involving TP53. Most cell lines (7/11) and a large fraction of tumor samples (10/25) showed TP53 rearrangements, in addition to somatic point mutations (6 patient samples, 1 cell line) and MDM2 amplifications (2 patient samples, 2 cell lines). The resulting inactivation of p53 was demonstrated by a deficiency of the radiation-induced DNA damage response. Thus, TP53 rearrangements are the major mechanism of p53 inactivation in osteosarcoma. Together with active MMBIR and MMEJ, this inactivation probably contributes to the exceptional chromosomal instability in these tumors. Although rampant rearrangements appear to be a phenotype of osteosarcomas, we demonstrate that among the huge number of probable passenger rearrangements, specific recurrent, possibly oncogenic, events are present. For the first time the genomic chaos of osteosarcoma is characterized so thoroughly and delivered new insights in mechanisms involved in osteosarcoma development and may contribute to new diagnostic and therapeutic strategies.

• Fykerud, Tone Aase; Knudsen, Lars Mørland; Totland, Max; Sørensen, Vigdis; Dahal-Koirala, Shiva & Lothe, Ragnhild A [Show all 8 contributors for this article] (2016). Mitotic cells form actin-based bridges with adjacent cells to provide intercellular communication during rounding. Cell Cycle. ISSN 1538-4101. 15(21), p. 2943–2957. doi: 10.1080/15384101.2016.1231280. Full text in Research Archive Show summary

  In order to achieve accurate chromosome segregation, eukaryotic cells undergo a dramatic change in morphology to obtain a spherical shape during mitosis. Interphase cells communicate directly with each other by exchanging ions and small molecules via gap junctions, which have important roles in controlling cell growth and differentiation. As cells round up during mitosis, the gap junctional communication between mitotic cells and adjacent interphase cells ceases. Whether mitotic cells use alternative mechanisms for mediating direct cell-cell communication during rounding is currently unknown. Here, we have studied the mechanisms involved in the remodeling of gap junctions during mitosis. We further demonstrate that mitotic cells are able to form actin-based plasma membrane bridges with adjacent cells during rounding. These structures, termed “mitotic nanotubes,” were found to be involved in mediating the transport of cytoplasm, including Rab11-positive vesicles, between mitotic cells and adjacent cells. Moreover, a subpool of the gap-junction channel protein connexin43 localized in these intercellular bridges during mitosis. Collectively, the data provide new insights into the mechanisms involved in the remodeling of gap junctions during mitosis and identify actin-based plasma membrane bridges as a novel means of communication between mitotic cells and adjacent cells during rounding.

• Sveen, Anita; Løes, Inger Marie; Alagaratnam, Sharmini; Nilsen, Gro; Høland, Maren & Lingjærde, Ole Christian [Show all 13 contributors for this article] (2016). Intra-patient inter-metastatic genetic heterogeneity in colorectal cancer as a key determinant of survival after curative liver resection. PLoS Genetics. ISSN 1553-7390. 12:e1006225(7). doi: 10.1371/journal.pgen.1006225. Full text in Research Archive
• Glaire, M.A.; Domingo, E.; Vermeulen, L.; Wezel, T van; Liefers, G.J. & Lothe, Ragnhild Adelheid [Show all 20 contributors for this article] (2016). POLE proofreading domain mutation defines a subset of immunogenic colorectal cancers with excellent prognosis. Annals of Oncology. ISSN 0923-7534. 27, p. vi152–vi152. doi: 10.1093/annonc/mdw370.09. Full text in Research Archive
• Domingo, Enric; Freeman-Mills, Luke; Rayner, Emily; Glaire, Mark; Briggs, Sarah & Vermeulen, Louis [Show all 35 contributors for this article] (2016). Somatic POLE proofreading domain mutation, immune response, and prognosis in colorectal cancer: a retrospective, pooled biomarker study. The Lancet Gastroenterology and Hepatology. ISSN 2468-1253. 1(3), p. 207–216. doi: 10.1016/S2468-1253(16)30014-0. Full text in Research Archive
• Kolberg, Matthias; Høland, Maren; Lind, Guro Elisabeth; Ågesen, Trude Holmeide; Skotheim, Rolf Inge & Hall, Kirsten Sundby [Show all 11 contributors for this article] (2015). Protein expression of BIRC5, TK1, and TOP2A in malignant peripheral nerve sheath tumours - A prognostic test after surgical resection. Molecular Oncology. ISSN 1574-7891. 9(6), p. 1129–1139. doi: 10.1016/j.molonc.2015.02.005. Full text in Research Archive
• Cekaite, Lina; Eide, Peter Andreas Wold; Lind, Guro Elisabeth; Skotheim, Rolf I. & Lothe, Ragnhild A (2015). MicroRNAs as growth regulators, their function and biomarker status in colorectal cancer. OncoTarget. 7(6), p. 6476–6505. doi: 10.18632/oncotarget.6390. Full text in Research Archive
• Mesquita, Diana; Barros-Silva, João D.; Santos, Joana; Skotheim, Rolf Inge; Lothe, Ragnhild A & Paulo, Paula [Show all 7 contributors for this article] (2015). Specific and redundant activities of ETV1 and ETV4 in prostate cancer aggressiveness revealed by co-overexpression cellular contexts. OncoTarget. 6(7), p. 5217–5236. doi: 10.18632/oncotarget.2847. Full text in Research Archive
• Brabrand, Sigmund; Johannessen, Bjarne; Axcrona, Ulrika; Kraggerud, Sigrid M.; Berg, Kaja Christine Graue & Bakken, Anne Cathrine [Show all 11 contributors for this article] (2015). Exome sequencing of bilateral testicular germ cell tumors suggests independent development lineages. Neoplasia. ISSN 1522-8002. 17(2), p. 167–174. doi: 10.1016/j.neo.2014.12.005. Full text in Research Archive
• Andresen, Kim; Boberg, Kirsten Muri; Vedeld, Hege Marie; Honne, Hilde; Jebsen, Peter Wilhelm & Hektoen, Merete [Show all 16 contributors for this article] (2015). Four DNA methylation biomarkers in biliary brush samples accurately identify the presence of cholangiocarcinoma. Hepatology. ISSN 0270-9139. 61(5), p. 1651–1659. doi: 10.1002/hep.27707. Full text in Research Archive
• Bruun, Jarle; Kolberg, Matthias; Ahlquist, Terje Cruickshank; Røyrvik, Ellen Christine; Nome, Torfinn & Leithe, Edward [Show all 18 contributors for this article] (2015). Regulator of chromosome condensation 2 identifies high-risk patients within both major phenotypes of colorectal cancer. Clinical Cancer Research. ISSN 1078-0432. 21(16), p. 3759–3770. doi: 10.1158/1078-0432.CCR-14-3294. Full text in Research Archive
• Hoff, Andreas Midbøe; Johannessen, Bjarne; Alagaratnam, Sharmini; Zhao, Sen; Nome, Torfinn & Løvf, Marthe [Show all 11 contributors for this article] (2015). Novel RNA variants in colorectal cancers. OncoTarget. 6(34), p. 36587–36602. doi: 10.18632/oncotarget.5500. Full text in Research Archive

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• Tasken, Kjetil; Russnes, Hege Elisabeth Giercksky; Aas, Eline; Bjørge, Line; Blix, Egil Støre & Ahlquist, Terje C. [Show all 116 contributors for this article] (2022). A national precision cancer medicine implementation initiative for Norway. Nature Medicine. ISSN 1078-8956. 28(5), p. 885–887. doi: 10.1038/s41591-022-01777-4. Full text in Research Archive
• Helland, Åslaug; Russnes, Hege Elisabeth Giercksky; Fagereng, Gro Live; Al-Shibli, Khalid Ibrahim; Andersson, Yvonne & Berg, Thomas [Show all 53 contributors for this article] (2022). Correction to: Improving public cancer care by implementing precision medicine in Norway: IMPRESS-Norway (Journal of Translational Medicine, (2022), 20, 1, (225), 10.1186/s12967-022-03432-5). Journal of Translational Medicine. 20(1). doi: 10.1186/s12967-022-03518-0. Full text in Research Archive
• Helland, Åslaug; Russnes, Hege Elisabeth Giercksky; Fagereng, Gro Live; Al-Shibli, Khalid Ibrahim; Andersson, Yvonne & Berg, Thomas [Show all 53 contributors for this article] (2022). Improving public cancer care by implementing precision medicine in Norway: IMPRESS-Norway. Journal of Translational Medicine. 20(1), p. 1–11. doi: 10.1186/s12967-022-03432-5. Full text in Research Archive Show summary

  Background Matching treatment based on tumour molecular characteristics has revolutionized the treatment of some cancers and has given hope to many patients. Although personalized cancer care is an old concept, renewed attention has arisen due to recent advancements in cancer diagnostics including access to high-throughput sequencing of tumour tissue. Targeted therapies interfering with cancer specific pathways have been developed and approved for subgroups of patients. These drugs might just as well be efficient in other diagnostic subgroups, not investigated in pharma-led clinical studies, but their potential use on new indications is never explored due to limited number of patients. Methods In this national, investigator-initiated, prospective, open-label, non-randomized combined basket- and umbrella-trial, patients are enrolled in multiple parallel cohorts. Each cohort is defined by the patient’s tumour type, molecular profile of the tumour, and study drug. Treatment outcome in each cohort is monitored by using a Simon two-stage-like ‘admissible’ monitoring plan to identify evidence of clinical activity. All drugs available in IMPRESS-Norway have regulatory approval and are funded by pharmaceutical companies. Molecular diagnostics are funded by the public health care system.

• Kryeziu, Kushtrim; Bergsland, Christian Holst; Guren, Tormod Kyrre; Sveen, Anita & Lothe, Ragnhild Adelheid (2022). Multiplex immunohistochemistry of metastatic colorectal cancer and ex vivo tumor avatars. Biochimica et Biophysica Acta. CR. Reviews on Cancer. ISSN 0304-419X. 1877(1). doi: 10.1016/j.bbcan.2022.188682. Full text in Research Archive
• Glaire, Mark; Domingo, Enric; Sveen, Anita; Bruun, Jarle; Nesbakken, Arild & Nicholson, George [Show all 16 contributors for this article] (2019). Correction: Tumour-infiltrating CD8+ lymphocytes and colorectal cancer recurrence by tumour and nodal stage (British Journal of Cancer, (2019), 121, 6, (474-482), 10.1038/s41416-019-0540-4). British Journal of Cancer. ISSN 0007-0920. 121(9). doi: 10.1038/s41416-019-0590-7. Full text in Research Archive
• Bruun, Jarle; Eide, Peter W.; Kryeziu, Kushtrim; Sveen, Anita; Murumagi, Astrid & Kolberg, Matthias [Show all 9 contributors for this article] (2017). Pharmacogenomic profiling to identify novel therapeutic strategies in colorectal cancer. Full text in Research Archive
• Oltedal, Satu; Kørner, Hartwig; Aasprong, Ole Gunnar; Hussain, Israr; Tjensvoll, Kjersti & Smaaland, Rune [Show all 12 contributors for this article] (2017). The Prognostic Relevance of Sentinel Lymph Node Metastases Assessed by PHGR1 mRNA Quantitation in Stage I-III Colon Cancer. Full text in Research Archive
• Jeanmougin, M; Håvik, Annette Bentsen; Cekaite, Lina; Brandal, Petter; Sveen, Anders & Meling, Torstein Ragnar [Show all 11 contributors for this article] (2016). Primary and secondary glioblastomas: global expression analysis of two histologically indistinguishable tumor subtypes. Full text in Research Archive
• Myklebost, Ola; Lothe, Ragnhild A; Gjertsen, Bjørn Tore & Lønning, Per Eystein (2015). Bør hver kreftpasient bli et eget forskningsprosjekt? Full text in Research Archive

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