Aurélie Beaufrère1, Stefano Caruso2, Julien Calderaro3, Nicolas Poté4, Jean-Charles Bijot5, Gabielle Couchy6, François Cauchy7, Valérie Vilgrain8, Jessica Zucman-Rossi9, Valérie Paradis10. 1. Université de Paris, Paris, France; APHP, Department of Pathology, Hôpital Beaujon, 100 boulevard du Général Leclerc, Clichy, 92110, France; INSERM UMR 1149, Centre de Recherche sur l'Inflammation, 16 rue Henri Huchard, Paris, 75018, France. 2. Centre de Recherche des Cordeliers, INSERM, Functional Genomics of Solid Tumors laboratory, F-75006 Paris, France. 3. Department of Pathology, Hôpital Henri Mondor, AP-HP, 51 Avenue du Maréchal de Lattre de Tassigny, Créteil, 94010, France. 4. Université de Paris, Paris, France; Department of Pathology, Hôpital Bichat, AP-HP.Nord, 46 Rue Henri Huchard, Paris, 75018, France. 5. Université de Paris, Paris, France; Department of Radiology, Hôpital Beaujon, AP-HP, 100 boulevard du Général Leclerc, Clichy, 92110, France. 6. Université de Paris, Paris, France; Centre de Recherche des Cordeliers, INSERM, Functional Genomics of Solid Tumors laboratory, F-75006 Paris, France. 7. Université de Paris, Paris, France; INSERM UMR 1149, Centre de Recherche sur l'Inflammation, 16 rue Henri Huchard, Paris, 75018, France; Department of HPB and Pancreatic surgery, Beaujon AP-HP, Clichy, 92110, France. 8. Université de Paris, Paris, France; INSERM UMR 1149, Centre de Recherche sur l'Inflammation, 16 rue Henri Huchard, Paris, 75018, France; Department of Radiology, Hôpital Beaujon, AP-HP, 100 boulevard du Général Leclerc, Clichy, 92110, France. 9. Université de Paris, Paris, France; Centre de Recherche des Cordeliers, INSERM, Functional Genomics of Solid Tumors laboratory, F-75006 Paris, France; Department of Oncology, Hopital Européen Georges Pompidou, AP-HP, F-75015, Paris, France. 10. Université de Paris, Paris, France; APHP, Department of Pathology, Hôpital Beaujon, 100 boulevard du Général Leclerc, Clichy, 92110, France; INSERM UMR 1149, Centre de Recherche sur l'Inflammation, 16 rue Henri Huchard, Paris, 75018, France. Electronic address: valerie.paradis@aphp.fr.
Abstract
BACKGROUND & AIMS: Microvascular invasion (MVI), a major risk factor for tumor recurrence after surgery in hepatocellular carcinoma (HCC), is only detectable by microscopic examination of the surgical specimen. We aimed to define a transcriptomic signature associated with MVI in HCC than can be applied to formalin-fixed paraffin-embedded (FFPE) biopsies for use in clinical practice. METHODS: To identify a gene expression signature related to MVI by using NanoString technology, we selected a set of 200 genes according to the literature and RNA-sequencing data obtained from a cohort of 150 frozen HCC samples previously published. We used 178 FFPE-archived HCC samples, including 109 surgical samples for the training set and 69 paired pre-operative biopsies for the validation set. In 14 cases of the training set, a paired biopsy was available and was also analyzed. RESULTS: We identified a 6-gene signature (ROS1, UGT2B7, FAS, ANGPTL7, GMNN, MKI67) strongly associated with MVI in the training set of FFPE surgical HCC samples, with 82% accuracy (sensitivity 82%, specificity 81%, AUC 0.82). The NanoString gene expression was highly correlated in 14 paired surgical/biopsy HCC samples (mean R: 0.97). In the validation set of 69 FFPE HCC biopsies, the 6-gene NanoString signature predicted MVI with 74% accuracy (sensitivity 73%, specificity 76%, AUC 0.74). Moreover, on multivariate analysis, the MVI signature was associated with overall survival in both sets (hazard ratio 2.29; 95% CI 1.03-5.07; p = 0.041). CONCLUSION: We defined a 6-gene signature that can accurately predict MVI in FFPE HCC biopsy samples, which is also associated with overall survival, although its survival impact must be confirmed by extensive study with further clinical data. LAY SUMMARY: Microvascular invasion, a major risk factor for tumor recurrence after surgery in hepatocellular carcinoma, is only detectable by microscopic examination of a surgical specimen. In this study, we defined a relevant surrogate signature of microvascular invasion in hepatocellular carcinoma that may be applied in clinical practice with routine tumor biopsy and integrated into the therapeutic strategy.
BACKGROUND & AIMS: Microvascular invasion (MVI), a major risk factor for tumor recurrence after surgery in hepatocellular carcinoma (HCC), is only detectable by microscopic examination of the surgical specimen. We aimed to define a transcriptomic signature associated with MVI in HCC than can be applied to formalin-fixed paraffin-embedded (FFPE) biopsies for use in clinical practice. METHODS: To identify a gene expression signature related to MVI by using NanoString technology, we selected a set of 200 genes according to the literature and RNA-sequencing data obtained from a cohort of 150 frozen HCC samples previously published. We used 178 FFPE-archived HCC samples, including 109 surgical samples for the training set and 69 paired pre-operative biopsies for the validation set. In 14 cases of the training set, a paired biopsy was available and was also analyzed. RESULTS: We identified a 6-gene signature (ROS1, UGT2B7, FAS, ANGPTL7, GMNN, MKI67) strongly associated with MVI in the training set of FFPE surgical HCC samples, with 82% accuracy (sensitivity 82%, specificity 81%, AUC 0.82). The NanoString gene expression was highly correlated in 14 paired surgical/biopsy HCC samples (mean R: 0.97). In the validation set of 69 FFPE HCC biopsies, the 6-gene NanoString signature predicted MVI with 74% accuracy (sensitivity 73%, specificity 76%, AUC 0.74). Moreover, on multivariate analysis, the MVI signature was associated with overall survival in both sets (hazard ratio 2.29; 95% CI 1.03-5.07; p = 0.041). CONCLUSION: We defined a 6-gene signature that can accurately predict MVI in FFPE HCC biopsy samples, which is also associated with overall survival, although its survival impact must be confirmed by extensive study with further clinical data. LAY SUMMARY: Microvascular invasion, a major risk factor for tumor recurrence after surgery in hepatocellular carcinoma, is only detectable by microscopic examination of a surgical specimen. In this study, we defined a relevant surrogate signature of microvascular invasion in hepatocellular carcinoma that may be applied in clinical practice with routine tumor biopsy and integrated into the therapeutic strategy.