Human cytokines activate JAK–STAT signaling pathway in porcine ocular tissue

Background: The JAK/STAT (Janus Tyrosine Kinase, Signal Transducers and Activators of Transcription) pathway is associated with cytokine or growth factor receptors and it is critical for growth control, developmental regulation and homeostasis. The use of porcine ocular cells as putative xenotransplants appears theoretically possible. The aim of this study was to investigate the response of various porcine ocular cells in vitro to human cytokines in regard to the activation of JAK-STAT signaling pathways. Methods: Porcine lens epithelial cells, pigmented iris epithelial cells and pigmented ciliary body cells were used in this study. These cells were isolated from freshly enucleated porcine eyes by enzymatic digestion. Cultured cells between passages 3–8 were used in all experiments. Electromobility shift assay (EMSA), proliferation assay, immunofluorescence staining and flow cytometry were used to evaluate the JAK-STAT signaling pathway in these cells. Results: JAK/STAT signaling pathways could be activated in porcine pigmented epithelial ciliary body cells, in pigmented iris epithelial cells and in lens epithelial cells in response to porcine and human interferons and cytokines. All cells showed very strong STAT1 activation upon stimulation with porcine interferon-gamma. Porcine ocular cells also respond to human cytokines; IFN-alpha induced strong activation of STAT1 in EMSA, flow cytometry and immunofluorescence experiments whereas activation of STAT3 was less strong in EMSA, but strong in flow cytometry and immunofluorescence. Human recombinant IL-6 activated STAT3 and human IL-4 activated STAT6. With the help of immunofluorescence assay and flow cytometry we observed nuclear localization of STAT proteins after activation of porcine ocular cells with cytokines and interferons. Human IFN-α had an inhibitory effect on porcine ocular cells in proliferation assays. Conclusion: Our study demonstrated that some types of human cytokines and interferon activate intracellular JAK-STAT signaling pathways in porcine ocular cells. We hypothesize that direct stimulation of the JAK-STAT pathway in porcine cells in response to human cytokines will lead to complications or failure, if pig-to-human ocular tissue xenotransplantation were to be carried out. For successful xenotransplantation among other obstacles there must be new approaches developed to regulate signaling pathways

Protein phosphatase 2A promotes hepatocellular carcinogenesis in the diethylnitrosamine mouse model through inhibition of p53

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. Most HCCs develop in cirrhotic livers. Alcoholic liver disease, chronic hepatitis B and chronic hepatitis C are the most common underlying liver diseases. Hepatitis C virus (HCV)-specific mechanisms that contribute to HCC are presently unknown. Transgenic expression of HCV proteins in the mouse liver induces an overexpression of the protein phosphatase 2A catalytic subunit (PP2Ac). We have previously reported that HCV-induced PP2Ac overexpression modulates histone methylation and acetylation and inhibits DNA damage repair. In this study, we analyze tumor formation and gene expression using HCV transgenic mice that overexpress PP2Ac and liver tissues from patients with HCC. We demonstrate that PP2Ac overexpression interferes with p53-induced apoptosis. Injection of the carcinogen, diethylnitrosamine, induced significantly more and larger liver tumors in HCV transgenic mice that overexpress PP2Ac compared with control mice. In human liver biopsies from patients with HCC, PP2Ac expression was significantly higher in HCC tissue compared with non-tumorous liver tissue from the same patients. Our findings demonstrate an important role of PP2Ac overexpression in liver carcinogenesis and provide insights into the molecular pathogenesis of HCV-induced HC

Proteogenomic characterization of hepatocellular carcinoma

We performed a proteogenomic analysis of hepatocellular carcinomas (HCCs) across clinical stages and etiologies. We identified pathways differentially regulated on the genomic, transcriptomic, proteomic and phosphoproteomic levels. These pathways are involved in the organization of cellular components, cell cycle control, signaling pathways, transcriptional and translational control and metabolism. Analyses of CNA-mRNA and mRNA-protein correlations identified candidate driver genes involved in epithelial-to-mesenchymal transition, the Wnt-β- catenin pathway, transcriptional control, cholesterol biosynthesis and sphingolipid metabolism. The activity of targetable kinases aurora kinase A and CDKs was upregulated. We found that CTNNB1 mutations are associated with altered phosphorylation of proteins involved in actin filament organization, whereas TP53 mutations are associated with elevated CDK1/2/5 activity and altered phosphorylation of proteins involved in lipid and mRNA metabolism. Integrative clustering identified HCC subgroups with distinct regulation of biological processes, metabolic reprogramming and kinase activation. Our analysis provides insights into the molecular processes underlying HCCs

Integrative proteogenomic characterization of hepatocellular carcinoma across etiologies and stages.

Proteogenomic analyses of hepatocellular carcinomas (HCC) have focused on early-stage, HBV-associated HCCs. Here we present an integrated proteogenomic analysis of HCCs across clinical stages and etiologies. Pathways related to cell cycle, transcriptional and translational control, signaling transduction, and metabolism are dysregulated and differentially regulated on the genomic, transcriptomic, proteomic and phosphoproteomic levels. We describe candidate copy number-driven driver genes involved in epithelial-to-mesenchymal transition, the Wnt-β-catenin, AKT/mTOR and Notch pathways, cell cycle and DNA damage regulation. The targetable aurora kinase A and CDKs are upregulated. CTNNB1 and TP53 mutations are associated with altered protein phosphorylation related to actin filament organization and lipid metabolism, respectively. Integrative proteogenomic clusters show that HCC constitutes heterogeneous subgroups with distinct regulation of biological processes, metabolic reprogramming and kinase activation. Our study provides a comprehensive overview of the proteomic and phophoproteomic landscapes of HCCs, revealing the major pathways altered in the (phospho)proteome

Understanding the role of water and tillage erosion from 239+240Pu tracer measurements using inverse modelling

Soil redistribution on arable land is a major threat for a sustainable use of soil resources. The majority of soil redistribution studies focus on water erosion, while wind and tillage erosion also induce pronounced redistribution of soil materials. Tillage erosion especially is understudied, as it does not lead to visible off-site damages. The analysis of on-site/in-field soil redistribution is mostly based on tracer studies, where radionuclide tracers (e.g. 137Cs, 239C240Pu) from nuclear weapon tests are commonly used to derive the erosion history over the past 50-60 years. Tracer studies allow us to determine soil redistribution patterns but integrate all types of soil redistribution processes and hence do not allow us to unravel the contribution of individual erosion processes. The aim of this study is to understand the contribution of water and tillage erosion leading to soil patterns found in a small hummocky ground moraine kettle hole catchment under intensive agricultural use. Therefore, 239C240Pu-derived soil redistribution patterns were analysed using an inverse modelling approach accounting for water and tillage erosion processes. The results of this analysis clearly point out that tillage erosion is the dominant process of soil redistribution in the study catchment, which also affects the hydrological and sedimentological connectivity between arable land and the kettle hole. A topographic change up to 17 cm (53 yr)1 in the eroded parts of the catchment is not able to explain the current soil profile truncation that exceeds the 239C240Pu-derived topographic change substantially. Hence, tillage erosion already started before the onset of intense mechanisation since the 1960s. In general, the study stresses the urgent need to consider tillage erosion as a major soil degradation process that can be the dominant soil redistribution process in sloped arable landscapes. © 2020 Georg Thieme Verlag. All rights reserved.ISSN:2199-3971ISSN:2199-398

Understanding the role of water and tillage erosion from 239+240Pu tracer measurements using inverse modelling

Soil redistribution on arable land is a major threat for a sustainable use of soil resources. The majority of soil redistribution studies focus on water erosion, while wind and tillage erosion also induce pronounced redistribution of soil materials. Tillage erosion especially is understudied, as it does not lead to visible off-site damages. The analysis of on-site/in-field soil redistribution is mostly based on tracer studies, where radionuclide tracers (e.g. 137Cs, 239C240Pu) from nuclear weapon tests are commonly used to derive the erosion history over the past 50-60 years. Tracer studies allow us to determine soil redistribution patterns but integrate all types of soil redistribution processes and hence do not allow us to unravel the contribution of individual erosion processes. The aim of this study is to understand the contribution of water and tillage erosion leading to soil patterns found in a small hummocky ground moraine kettle hole catchment under intensive agricultural use. Therefore, 239C240Pu-derived soil redistribution patterns were analysed using an inverse modelling approach accounting for water and tillage erosion processes. The results of this analysis clearly point out that tillage erosion is the dominant process of soil redistribution in the study catchment, which also affects the hydrological and sedimentological connectivity between arable land and the kettle hole. A topographic change up to 17 cm (53 yr)1 in the eroded parts of the catchment is not able to explain the current soil profile truncation that exceeds the 239C240Pu-derived topographic change substantially. Hence, tillage erosion already started before the onset of intense mechanisation since the 1960s. In general, the study stresses the urgent need to consider tillage erosion as a major soil degradation process that can be the dominant soil redistribution process in sloped arable landscapes. © 2020 Georg Thieme Verlag. All rights reserved.ISSN:2199-3971ISSN:2199-398

Human cytokines activate JAK-STAT signaling pathway in porcine ocular tissue

BACKGROUND: The JAK/STAT (Janus Tyrosine Kinase, Signal Transducers and Activators of Transcription) pathway is associated with cytokine or growth factor receptors and it is critical for growth control, developmental regulation and homeostasis. The use of porcine ocular cells as putative xenotransplants appears theoretically possible. The aim of this study was to investigate the response of various porcine ocular cells in vitro to human cytokines in regard to the activation of JAK-STAT signaling pathways. METHODS: Porcine lens epithelial cells, pigmented iris epithelial cells and pigmented ciliary body cells were used in this study. These cells were isolated from freshly enucleated porcine eyes by enzymatic digestion. Cultured cells between passages 3-8 were used in all experiments. Electromobility shift assay (EMSA), proliferation assay, immunofluorescence staining and flow cytometry were used to evaluate the JAK-STAT signaling pathway in these cells. RESULTS: JAK/STAT signaling pathways could be activated in porcine pigmented epithelial ciliary body cells, in pigmented iris epithelial cells and in lens epithelial cells in response to porcine and human interferons and cytokines. All cells showed very strong STAT1 activation upon stimulation with porcine interferon-gamma. Porcine ocular cells also respond to human cytokines; IFN-alpha induced strong activation of STAT1 in EMSA, flow cytometry and immunofluorescence experiments whereas activation of STAT3 was less strong in EMSA, but strong in flow cytometry and immunofluorescence. Human recombinant IL-6 activated STAT3 and human IL-4 activated STAT6. With the help of immunofluorescence assay and flow cytometry we observed nuclear localization of STAT proteins after activation of porcine ocular cells with cytokines and interferons. Human IFN-alpha had an inhibitory effect on porcine ocular cells in proliferation assays. CONCLUSION: Our study demonstrated that some types of human cytokines and interferon activa intracellular JAK-STAT signaling pathways in porcine ocular cells. We hypothesize that direct stimulation of the JAK-STAT pathway in porcine cells in response to human cytokines will lead to complications or failure, if pig-to-human ocular tissue xenotransplantation were to be carried out. For successful xenotransplantation among other obstacles there must be new approaches developed to regulate signaling pathways