Overexpression and alternative splicing of NF-YA in breast cancer

NF-Y is a CCAAT-binding trimeric transcription factor, whose regulome, interactome and oncogenic potential point to direct involvement in cellular transformation. Yet little is known about the levels of NF-Y subunits in tumors. We focused on breast carcinomas, and analyzed RNA-Seq datasets of TCGA and 54 BRCA cell lines at gene and isoforms level. We partitioned all tumors in the four major subclasses. NF-YA, but not histone-fold subunits NF-YB/NF-YC, is globally overexpressed, correlating with the proliferative Ki67 marker and a common set of 840 genes, with cell-cycle, metabolism GO terms. Their promoters are enriched in NF-Y, GC-rich and E2F sites. Surprisingly, there is an isoform switch, with the "short" isoform -NF-YAs- becoming predominant in tumors. E2F genes are also overexpressed in BRCA, but no switch in isoforms is observed. In Basal-like Claudinlow cell lines and tumors, expression of NF-YAl -long- isoform is high, together with 11 typical EMT markers and low levels of basal Keratins. Analysis of Progression-Free-Intervals indicates that tumors with unbalance of NF-YA isoforms ratios have worst clinical outcomes. The data suggest that NF-YA overexpression increases CCAAT-dependent, pro-growth genes in BRCA. NF-YAs is associated with a proliferative signature, but high levels of NF-YAl signal loss of epithelial features, EMT and acquisition of a more aggressive behavior in a subset of Claudinlow Basal-like tumors

Experimental and theoretical investigations in the stereoselective synthesis of P-stereogenic phosphine oxides

International audienceAn efficient enantioselective strategy for the synthesis of variously substituted phosphine oxides has been developed, incorporating the use of (1S,2S)-2-aminocyclohexanol as the chiral auxiliary. The method relies on three key steps: 1) Highly diastereoselective formation of PV oxazaphospholidine, rationalized by a theoretical study; 2) highly diastereoselective ring-opening of the oxazaphospholidine oxide with organometallic reagents that takes place with inversion of configuration at the P atom; 3) enantioselective synthesis of phosphine oxides by cleavage of the remaining P[BOND]O bond. Interestingly, the use of a PIII phosphine precursor afforded a P-epimer oxazaphospholidine. Hence, the two enantiomeric phosphine oxides can be synthesized starting from either a PV or a PIII phosphine precursor, which constitutes a clear advantage for the stereoselective synthesis of sterically hindered phosphine oxides

"Biological identikit": development of a SNPs-panel for the analysis of forensic DNA phenotyping and ancestry

The need to identify victims of mass disasters and perpetrators of crimes is fundamental for humanitarian, ethical and legal reasons. Individuals, whose genetic profiles are not yet known to investigators, cannot be identified by standard forensic DNA analysis. To date, technological progress allows for the analysis of Forensic DNA Phenotyping (FDP) which provides for the definition of Externally Visible Features (EVCs); and the analysis of the biogeographical ancestry (BGA), which allows to define the ancestral origin. The aim of this study was to evaluate the potential of a new panel of about 900 SNPs in predicting phenotypic traits and biogeographical origin. Recent biological material was analysed and subsequently the analysis was extended to include evidence found at the crime scene or extracted DNA, both preserved for long periods of time. The goal was to create a "biological identikit", useful for determining the phenotypic characteristics of unknown corpses and directing investigators to alleged victims or relatives or in a crime to narrow the circle of suspects. The panel consists of 41 SNPs for phenotyping and 850 SNPs for ancestry. It was tested on 8 recent samples and 26 samples extracted and stored for a long time. All the 891 SNPs were successfully incorporated into a single two-step multiplex PCR reaction using the IonAmpliSeq ™ Library Plus and applied for massive parallel sequencing with the Ion S5 platform using up to 0.05 ng / µl of DNA. The analysis of the results was carried out with an in-house predictive algorithm and by consulting 20 databases containing population frequencies. By comparing the results obtained with identikit or video-photographic surveys, it was possible to predict the phenotype and ancestry with an accuracy more than 90%. Although these new markers are unable to identify an individual, they can be a valuable investigative tool useful for personal identification