have recently reported that rapamycin, reducing Plasminogen activator inhibitor 1 1 expression, was able to decrease extracellular matrix CC5013 deposition in all renal compartments of patients with chronic allograft nephropathy. On the other hand, high concentrations of EVE, through a massive mTORC1 inhibition, may lead to a down regulation of S6K and a subsequent hyper activation of mTORC2 that, sustaining the phosphorylation of AKT at S473, could induce a feedback loop that stimulates PI3K AKT signaling activating the cellularmolecular machinery leading to renal fibrosis. In particular AKT, once activated, could induce, through the inhibition of Glycogen synthase kinase 3, the nuclear translocation of B catenin which stimulates the expression of EMT associated genes.

Our data Inhibitors,Modulators,Libraries confirmed that the knock down of AKT can control the activation of EMT program. This confirms previous results from pharmacodynamic analysis of cancer patient derived tumor material showed increased AKT S473 phosphorylation in some cases after treatment with doses and schedules of EVE defined as biologically optimal through Inhibitors,Modulators,Libraries pharmacokineticpharmacody namic modeling of preclinical and phase I data. Additionally, we emphasized that, as previously dem onstrated, HPSE has a pivotal role in the aforementioned pathway. In fact, the silencing of this enzyme in our cellular model reversed the activation of the EMT. Heparanase is an endo B D glucuronidase that cleaves heparan sulfate side chains at a limited number of sites, hence participates in ECM degradation and remodeling.

The degradation of several con stituents Inhibitors,Modulators,Libraries of the ECM, including heparan sulfate proteo glycans, promotes the release of growth factors such as FGF 2. Moreover, we previously shown that HPSE is necessary to sustain the PI3KAKT pathway mediated by FGF 2 which induces the expression of mesenchymal markers SMA Inhibitors,Modulators,Libraries and Vimentin, leads to degrad ation of the basement membrane by means of the secre tion of matrix metalloproteinases and increases cell motility. The heparanase expression is finely reg ulated by transcription factor, DNA methylation and vari ous endogenous molecules. Finally in order to find new elements involved in EVE induced EMT, we analyzed the differences in the tran scriptomic profile between HK 2 EVE treated cells and controls. Our study was performed using a microarray technology able to evaluate simultan eously the expression of more than 30,000 genes.

Inhibitors,Modulators,Libraries How ever, to take together full advantage of the opportunities offered by this high throughput method, it is necessary to manage, integrate and interpret a huge amount of data correctly. Thus, we decided to use a pathway analysis to focus our research on candidate genes known to be associated with EMT in order to reduce the false positive rate and the puzzling factors not directly associated with the aims of our research. Different statistical algorithms identified two genes significantly up regulated by this drug.