Fig. 2
Prognostic value of circulating microRNAs and TP53 mutational status of matched RMs. (a) A 4-prognostic microRNA signature was assessed on RMs collected at disease onset (RMp) or relapse (RMr) and compared with tumor relapse. Patients were stratified according to their clinical outcome. Relative expression of microRNAs is shown. Dynamics of TP53 VAF is indicated. (b) The same microRNA signature was assessed on tissues from pt#3 (left) and pt#2 (right). For pt#3, the RM of the first primitive tumor (RMp#1) developed on the palatine tonsil, on which he recurred, was compared with the RM of the second primitive tumor (RMp#2) on floor of the mouth, on which the patient has ever not recurred. For pt#2, RMp has been compared with the PHE lesion, developed one year before the first recurrence. Relative expression of microRNAs is shown. Dynamics of TP53 VAF is indicated. (c-d) Box plot and KM analysis showing the diagnostic and prognostic value of our circulating microRNA signature to early predict local recurrence. microRNAs expression of sera collected at 1 day or 15 days post-surgery has been normalized to microRNAs expression of matched pre-surgery sera. (e-f) KM and ROC curve analyses according to mutational status of RMs and microRNAs signature expression at 1 day (left) or 15 days post-surgery (right). For each KM, HR value and the relative confident interval (CI) 95% has been indicated. (g) Supervised clustering (left) analysis representing the expression of the 3 prognostic microRNAs in normal tissues from 13 HNSCC patients cultured with CM from Cal27 cells according to patient’s outcome. Colors represent folds of modulation of CM vs RPMI. Box plot (right) representing the expression level of miR-21-3p and miR-21-5p significantly (p=0.02) up-regulated in histologically tumor-free tissues from n=5 recurrent patients vs n=8 patients with no evidence of disease (NED) for at least 36 months cultured with CM as compared to the same tissues cultured with RPMI. (h) Supervised clustering (left) analysis representing the expression of the 3 prognostic microRNAs in normal tissues from 13 HNSCC patients cultured with RPMI medium according to their clinical outcome. Box plot (right) showing miR-96-5p up-regulation (p=0.04) in histologically tumor-free tissues from n=5 recurrent patients vs n=8 patients with NED for at least 36 months, cultured in the presence of RPMI. Raw data of miRNAs expression and the relative patient outcome are available in Suppl. Tables, sheet 5-6. HR: hazard ratio; NED: no evidence of the disease; Rec: recurrence; RFS: recurrence-free survival.