An altered metabolism is involved in the development of clear cell - renal cell carcinoma (ccRCC), and in this tumor many altered genes play a fundamental role in controlling cell metabolic activities. We delineated a large-scale metabolomic profile of human ccRCC, and integrated it with transcriptomic data to connect the variations in cancer metabolism with gene expression changes. Moreover, to better analyze the specific contribution of metabolic gene alterations potentially associated with tumorigenesis and tumor progression, we evaluated the transcription profile of primary renal tumor cells. Untargeted metabolomic analysis revealed a signature of an increased glucose uptake and utilization in ccRCC. In addition, metabolites related to pentose phosphate pathway were also altered in the tumor samples in association with changes in Krebs cycle intermediates and related metabolites. We identified NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 4-like 2 (NDUFA4L2) as the most highly expressed gene in renal cancer cells and evaluated its role in sustaining angiogenesis, chemoresistance, and mitochondrial dysfunction. Finally, we showed that silencing of NDUFA4L2 affects cell viability, increases mitochondrial mass, and induces ROS generation in hypoxia.

Epigenetic mechanisms, such as DNA methylation, affect tubular maladaptive response after Acute Kidney Injury (AKI) and accelerate renal aging. Upon ischemia/reperfusion (I/R) injury, Complement activation leads to C5a release that mediates damage; however, little is known about the effect of C5a-C5a Receptor (C5aR) interaction in Renal Tubular Epithelial Cells (RTEC). Through a whole-genome DNA methylation analysis in cultured RTEC, we found that C5a induced aberrant methylation, particularly in regions involved in cell cycle control, DNA damage and Wnt signaling. The most represented genes were BCL9 , CYP1B1 and CDK6 . C5a stimulation of RTEC led to up-regulation of SA-β Gal and cell cycle arrest markers such as p53 and p21. C5a increased also IL-6 , MCP-1 and CTGF gene expression, consistent with SASP development. In accordance, in a swine model of renal I/R injury, we found the increased expression of Wnt4 and βcatenin correlating with SA-β Gal, p21, p16 and IL-6 positivity. Administration of Complement Inhibitor (C1-Inh), antagonized SASP by reducing SA-β Gal, p21, p16, IL-6 and abrogating Wnt4/βcatenin activation. Thus, C5a affects the DNA methylation of genes involved in tubular senescence. Targeting epigenetic programs and Complement may offer novels strategies to protect tubular cells from accelerated aging and to counteract progression to Chronic Kidney Disease

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Background: Solid organ transplantation is an available therapeutic option for cystic fibrosis (CF) patients without lung transplantation. However, the use of immunosuppressive agents may cause severe adverse events. In particular, patients treated with mTOR-inhibitors (mTOR-I) may aggravate pulmonary complications. It has been recently described that these drugs may induce epithelial to mesenchymal transition (EMT) of airway cells. Objective: The purpose of this study was to evaluate the effects of mTOR-I on primary bronchial epithelial cells carrying F508del. Materials and Methods: Human bronchial epithelial cells homozygous for F508del were treated with 5 and 100nM EVE for 24 hours and their RNA was extracted and hybridized to the Human HT-12 v3 Expression BeadChip (Illumina). Microarray results were validated by Real-Time PCR. Transepithelial resistance was measured by Millicell-ERS ohmmeter. Results: High dosage EVE induced a significant up-regulation of 48 genes and a down-regulation of 14 genes. After pathway analysis by GSEA, we found that most of them were implicated in the inflammatory and pro-fibrotic pathways. Real-Time PCR confirmed that 100nM EVE was able to up-regulate some identified genes (IL-1 α IL-8, Pim-1) as well as pro-fibrotic elements ( α -SMA, connective tissue growth factor and metalloproteinase-12). Additionally, high dosage of EVE was also able to reduce the transepithelial resistance. In contrast, a lower level of EVE did not produce similar effects. Conclusion: Although performed in vitro, our study suggested that in solid organ transplant recipients with CF without a lung transplant, mTOR-I should be used at a low dosage to reduce its contribution to pulmonary inflammation and fibrosis.