MicroRNA expression profiling with a droplet digital PCR assay enables molecular diagnosis and prognosis of cancers of unknown primary. Issue 10 (23rd June 2021)

• Record Type: Journal Article
• Authors: Laprovitera, Noemi; Riefolo, Mattia; Porcellini, Elisa; Durante, Giorgio; Garajova, Ingrid; Vasuri, Francesco; Aigelsreiter, Ariane; Dandachi, Nadia; Benvenuto, Giuseppe; Agostinis, Federico; Sabbioni, Silvia; Berindan Neagoe, Ioana; Romualdi, Chiara; Ardizzoni, Andrea; Trerè, Davide; Pichler, Martin; D'Errico, Antonietta; Ferracin, Manuela
• Is Part Of: Molecular oncology. Volume 15:Issue 10(2021)
• Page Start: 2732
• Page End: 2751
• Publication Date: 2021-06-23
• Subjects: cancer of unknown primary -- droplet digital PCR -- metastasis -- microRNAs -- molecular diagnostics; Cancer -- Molecular aspects -- Periodicals; 616.994005
• DOI: 10.1002/1878-0261.13026
• Languages: English
• ISSNs: 1574-7891

Abstract

Abstract : Metastasis is responsible for the majority of cancer‐related deaths. Particularly, challenging is the management of metastatic cancer of unknown primary site (CUP), whose tissue of origin (TOO) remains undetermined even after extensive investigations and whose therapy is rather unspecific and poorly effective. Molecular approaches to identify the most probable TOO of CUPs can overcome some of these issues. In this study, we applied a predetermined set of 89 microRNAs (miRNAs) to infer the TOO of 53 metastatic cancers of unknown or uncertain origin. The miRNA expression was assessed with droplet digital PCR in 159 samples, including primary tumors from 17 tumor classes (reference set) and metastases of known and unknown origin (test set). We combined two different statistical models for class prediction to obtain the most probable TOOs: the nearest shrunken centroids approach of Prediction Analysis of Microarrays (PAMR) and the least absolute shrinkage and selection operator (LASSO) models. The molecular test was successful for all formalin‐fixed paraffin‐embedded samples and provided a TOO identification within 1 week from the biopsy procedure. The most frequently predicted origins were gastrointestinal, pancreas, breast, lung, and bile duct. The assay was applied also to multiple metastases from the same CUP, collected from different metastatic sites: The predictions showed a strong agreement, intrinsically validating our assay. The final CUPs' TOO prediction was compared with the clinicopathological hypothesis of primary site. Moreover, a panel of 13 miRNAs proved to have prognostic value and be associated with overall survival in CUP patients. Our study demonstrated that miRNA expression profiling in CUP samples could be employed as diagnostic and prognostic test. Our molecular analysis can be performed on request, concomitantly with standard diagnostic workup and in association with genetic profiling, to offer valuable indications about the possible primary site, thereby supporting treatment decisions. Abstract : Cancer of unknown primary site (CUP) patients suffer the burden of a metastatic disease whose site of origin is unidentifiable, even after intensive clinical investigations. The lack of a recognized primary site limits patients' treatment options. In this manuscript, we present a molecular assay, based on digital miRNA profiling, that allowed the prediction of the primary site of 53 CUPs.