A broad array of anticancer activities of the immunosuppressive drug mycophenolic acid (MPA) is suggested by pathway analyses of gene expression and validated by proteomic and functional assays
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ABSTRACT: Gene expression microarray was performed to assess the global changes coupled to targeted validation of proteomic profiling to reveal the broad anticancer activities of the immunosuppressive drug Mycophenolic acid (MPA) and elucidate the molecular mechanisms underlying these activities. We conducted gene expression microarray experiments on AGS cancers cells treated with Mycophenolic acid (MPA). A concentration of 2ug/ml was selected to treat AGS cells and the treated cells were harvested at 0, 12, 24, 48 and 72 hours of culture with the drug. Gene expression analyses were carried out using Illumina Human 12 Beadchips.
Project description:Gene expression microarray was performed to assess the global changes coupled to targeted validation of proteomic profiling to reveal the broad anticancer activities of the immunosuppressive drug Mycophenolic acid (MPA) and elucidate the molecular mechanisms underlying these activities.
Project description:Mycophenolic acid (MPA), effectively promoted ES cell pancreatic differentiation with a concomitant reduction of neuronal cells. The targets of MPA were investigated by microarray analysis of cells treated on day 5 with MPA and harvested on day 6 and 8.
Project description:Mycophenolic acid (MPA), effectively promoted ES cell pancreatic differentiation with a concomitant reduction of neuronal cells. The targets of MPA were investigated by microarray analysis of cells treated on day 5 with MPA and harvested on day 6 and 8. Cells were harvested for RNA extraction and hybridization on Affymetrix microarrays. We performed gene expression profiles of differentiated mouse ES cells treated with or without 2 microM MPA at day5, 6, and 8
Project description:To know the differences of decidualization induced by different stimuli, we compared the mRNA expression profiles of decidualized ESCs stimulated by cAMP or cAMP+MPA for 4 days, or by MPA or E2+MPA for 14 days.
Project description:Mycophenolic Acid (MPA) is the active component of the immunosuppressant Mycophenolate Mofetil, a potent inhibitor of the inosine monophosphate dehydrogenase. Direct effects of MPA on podocytes remain largely unknown. In order to elucidate genes and pathways affected by the drug, cultured murine podocytes exposed to MPA were subjected to RNA sequencing (RNASeq) analysis. Untreated samples served as controls. The RNASeq of MPA treated podocytes identified 351 significantly affected genes (padj < 0.05; 130 downregulated / 221 upregulated). Gene Ontology-Term enrichment analysis outlined two major groups of terms of particular interest, namely actin associated terms and terms related to inflammatory cell death. In conclusion, MPA treatment has a substantial effect on the transcriptome of podocytes. Analysis of the sequencing data revealed several non-immune cell dependent areas in which MPA possibly has a favorable effect on podocytes.
Project description:Mycophenolic acid (MPA), an immunosuppressive drug widely used in kidney transplantation, has been suggested to have anti-fibrotic effects. To analyze at a genomic level these effects, we prospectively studied a group of stable kidney transplant recipients (n=35) on cyclosporine (CyA) and azathioprine treatment. Twenty patients were converted from azathioprine to MPA (MPA group) and 15 patients continued on azathioprine (AZA group). RNA was extracted by peripheral blood mononuclear cells at baseline and 3 months thereafter. Genomic analysis, performed on 5 randomly-selected MPA patients, revealed that 17 genes discriminated the transcriptomic profile after conversion. Neutral endopeptidase (NEP), an enzyme degrading angiotensin-II, was the most significant up-regulated gene. NEP expression level was inversely correlated to proteinuria at baseline and after conversion. Immunohistochemistry on graft biopsy of 33 independent patients demonstrated higher glomerular and tubular NEP protein expression in CyA+MPA (n=13) compared to CyA+AZA (n=12) and CyA alone (n=8). Glomerular NEP levels were inversely correlated to proteinuria and glomerulosclerosis. Tubular NEP expression was inversely correlated to interstitial fibrosis. Incubation of proximal tubular cells with MPA led to a dose- and time-dependent increase of NEP gene expression. The direct influence of MPA on NEP expression may suggest a novel therapeutic effect of this drug.
Project description:Hydrostatic pressure is one of the physical factors affecting cellular physiology. Hydrostatic pressure of a few hundred MPa decreases the viability of yeast cells, and pressure of a few tens MPa decreases the growth rate. To understand the effect of hydrostatic pressure, we employed yeast, Saccharomyces cerevisiae, DNA microarrays and analyzed genome-wide mRNA expression profiles under hydrostatic pressures. In this experiment, we selected a hydrostatic pressure of 30 MPa at 25 degrees C because yeast cells are able to grow with this condition. Keywords: stress response
Project description:Hydrostatic pressure is one of the physical factors affecting cellular physiology. Hydrostatic pressure of a few hundred MPa decreases the viability of yeast cells, and pressure of a few tens MPa decreases the growth rate. To understand the effect of hydrostatic pressure, we employed yeast, Saccharomyces cerevisiae, DNA microarrays and analyzed genome-wide mRNA expression profiles under hydrostatic pressures. In this experiment, we selected a hydrostatic pressure of 40 MPa at 25 degrees C because the condition is not lethal for yeast cells but the growth was suppressed. Keywords: stress response
Project description:Mycophenolic acid (MPA) is a potent inhibitor of the inosine monophosphate dehydrogenase and commonly used as an immunosuppressive drug in transplantation. MPA inhibits proliferation of both T- and B-lymphocytes by guansoin depletion. Since fibroblasts rely on the de novo synthesis of guanosin nucleotides, it is assumed that MPA interacts with fibroblasts causing an increased frequency of wound healing problems. We show a downregulation of the cytoskeletal proteins actin, vinculin and tubulin in human dermal fibroblasts exposed to pharmacologic doses of MPA using microarray technology and western blot. This reduction in protein content is accompanied by a substantial derangement of the cytoskeleton in MPA-treated fibroblasts as documented by confocal microscopy. The dysfunctional fibroblast growth was validated by scratch test documenting impaired migrational capacity. The results of the cultured dermal fibroblasts were applied to skin biopsies of renal transplant recipients. Skin biopsies of patients treated with MPA expressed less tubulin and actin as compared to control biopsies which could explain potential wound healing problems post transplantation. The perspective of MPA-induced cytoskeletal dysfunction may go beyond wound healing disturbances and has potential beneficial effects on (renal) allografts with respect to scarring. Keywords: Timecourse and MPA and/or Guanosin response