Effective biomarkers predictive of the response to treatments are key for precision medicine. This study identifies the staining pattern of the centromeric histone 3 variant, CENP-A, as a predictive biomarker of locoregional disease curability by chemoradiation therapy. We compared by imaging the subnuclear distribution of CENP-A in normal and tumoral tissues, and in a retrospective study in biopsies of 62 locally advanced head and neck squamous cell carcinoma (HNSCC) patients treated by chemoradiation therapy. We looked for predictive factors of locoregional disease control and patient’s survival, including CENP-A patterns, Ki67, HPV status and anisokaryosis. In different normal tissues, we reproducibly found a CENP-A subnuclear pattern characterized by CENP-A clusters both localized at the nuclear periphery and regularly spaced. In corresponding tumors, both features are lost. In locally advanced HNSCC, a specific CENP-A pattern identified in pretreatment biopsies predicts definitive locoregional disease control after chemoradiation treatment in 96% (24/25) of patients (OR = 17.6 CI 95% [2.6; 362.8], p = 0.002), independently of anisokaryosis, Ki67 labeling or HPV status. The characteristics of the subnuclear pattern of CENP-A in cell nuclei revealed by immunohistochemistry could provide an easy to use a reliable marker of disease curability by chemoradiation therapy in locally advanced HNSCC patients.

The PIWI proteins emerging in the development of human cancers, edify PIWI-piRNA ribonucleoproteic complexes acting as pivotal regulators of genome integrity, differentiation and homeostasis. The aim of this study is to analyze the four PIWILs gene expression in invasive breast carcinomas (IBCs): at RNA level using quantitative RT-PCR (n = 526) and protein level using immunohistochemistry (n = 150). In normal breast tissue, PIWILs 2 and 4 were solely expressed, whereas an abnormal emergence of PIWIL1 and 3 was observed in respectively 30% and 6% of IBCs. Conversely, PIWIL2 was underexpressed in 48.3% and PIWIL4 downregulated in 43.3% of IBCs. Significant positive associations were observed between PIWIL4 underexpression, HR+ status and HR+ ERBB2+ molecular subtype and PIWIL2 underexpression, PR- status, ERBB2- status and molecular subtype. Similar patterns of PIWIL deregulation were observed in a multitumoral panel, suggesting a generic mechanism in most cancers. PIWIL2-4 underexpression was mainly regulated at epigenetic or post-transcriptional levels. PIWIL2 underexpression was significantly associated with DNA methylation and strong cytotoxic immune response. PIWIL2-4 were mainly associated with genes implicated in cell proliferation. As a result of this study, characterization of the PIWIL-piRNA pathway in IBCs opens interesting therapeutic perspectives using piRNAs, hypomethylating drugs, checkpoints immunotherapies and anti-PIWIL 1–3 antibodies.

Introduction To analyse the Epstein Barr Virus (EBV) part in biological and clinical characteristics of patients treated for a classical Hodgkin lymphoma (cHL) in France. Methods Biopathological datas were centrally reviewed in 299 patients with cHL. Patients were treated in or according to the Euronet PHL-C1 trial between November 2008 to February 2013. Results Median age at diagnosis was 14 [3-18], F/M ratio was 0,84; 0,47 between 3 to 10 years, and 0,9 from 11 to 18. CHL subtypes were nodular sclerosis (cHL NS) 265/299 (88,6%), mixed cellularity (cHL MC) 22/299 (7,4%), lymphocyte rich 2/299 (0,7%) and 10/299 (3,3%) were unclassified. 68/299 (22,7%) present a cHL EBV+, significantly more frequent in 3-10 years patients n=17/34 (50%), than in >11years n=51/265 (19,2%) p<0,001, and in cHL MC subtype n=15/22 (68,2%), than in cHL NS subtype n=49/265 (18,5%) p<0,001. EBV serology was recorded in 100/299 (33,4%) cases. Anti VCA-IgG were positive in 70/100 (70%) cases, anti EBNA-IgG were positive in 61/100 (61%) cases, and anti VCA-IgM were positive in 5/100 (5%) cases. In EBV+ cases, 22/23 (95,7%) were VCA-IgG+ (p=0,002), 19/23 (82,6%) were EBNA-IgG+ (p=0,01), and 2/23 (8,7%) were VCA-IgM+ (p=0,3). EBV PCR was recorded in 108/299 (36,1%) cases, and was positive in 22/108 (20,4%). In EBV+ cases, 13/28 (46,4%) had a PCR+ (p<0,001). No significant difference in sex distribution (p=0,12) and staging disease (p=0,84) was present in cHL EBV+/- . Immunostaining was: 284/284 (100%) CD30+, (no data for 15 patients); CD15 was + 242/284 (85,2%) cases and 55/62 (88,7%)in cHL EBV+ (p=0,38). CD20 was + 62/287 cases (21,6%) and 15/63 (23,8%) in cHL EBV+ (p=0,23). PAX5 was + 211/255 (82,7%) with cHL EBV+ 44/52 (84,6%) (p=0,69). Overall, no significant differences between immunostaining in cHL EBV+and negative distribution has been highlighted. No significant difference in overall survival (p=0,35) and event free survival (p=0,9) has been raised between EBV positive and negative population. Conclusion In this cohort of 299 French children and adolescents with cHL, cHL MC subtype is present in only 12.4%. EBV cHL represent 22,7% of our cohort and is significantly associated with young age and cHL MC, without survival or relapse impact. Although EBV in cHL is well-known, EBV immune role in HL pathology need further research.