Proteomic analysis of morphologically changed tissues after prolonged dexamethasone treatment

Prolonged dexamethasone (Dex) administration leads to serious adverse and decrease brain and heart size, muscular atrophy, hemorrhagic liver, and presence of kidney cysts. Herein, we used an untargeted proteomic approach using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for simultaneous identification of changes in proteomes of the major organs in Sprague-Dawley (SD rats post Dex treatment. The comparative and quantitative proteomic analysis of the brain, heart, muscle, liver, and kidney tissues revealed differential expression of proteins (n = 190, 193, 39, 230, and 53, respectively) between Dex-treated and control rats. Functional network analysis using ingenuity pathway analysis (IPA revealed significant differences in regulation of metabolic pathways within the morphologically changed organs that related to: (i) brain-cell morphology, nervous system development, and function and neurological disease; (ii) heart-cellular development, cellular function and maintenance, connective tissue development and function; (iii) skeletal muscle-nucleic acid metabolism, and small molecule biochemical pathways; (iv) liver-lipid metabolism, small molecular biochemistry, and nucleic acid metabolism; and (v) kidney-drug metabolism, organism injury and abnormalities, and renal damage. Our study provides a comprehensive description of the organ-specific proteomic profilesand differentially altered biochemical pathways, after prolonged Dex treatement to understand the molecular basis for development of side effects

Behavior of the density current in a semi-closed water body and tidal river

研究科: 千葉大学大学院自然科学研究科博士(学術)報告番号:甲第学287

Cancer proteomics : characterisation of protein expression in human epithelial tumours

Defining differences in protein expression between tumour cells of various degrees of malignancy is a first step in the development of markers for tumour diagnosis. In this thesis, the expression of multiple proteins in benign and malignant human ovarian, breast and prostate tumours was examined using two-dimensional gel electrophoresis (2-DE). We were able to show a similar pattern of expression of a set of ten proteins between benign and malignant cells in all three tumour types examined. Cytokeratins and high molecular weight tropomyosins were consistently down-regulated in carcinomas, whereas stress proteins (HSP90, HSP60 and calreticulin) were upregulated. This finding suggests a high degree of homology in the expression of these proteins among different tumour types of epithelial origin. An attempt to apply principal component analysis of quantitative 2-DE data for diagnosis of ovarian cancer is presented. Data of the expression of 170 polypeptides was compiled from 22 tumours and used to construct a model for classification into benign, borderline and malignant ovarian tumours. When the model was tested using 18 tumours, 11 tumours (61%) were correctly classified. We were encouraged by this result and intend to increase the number of tumours used to construct the model. Future work will show whether it is possible to accurately classify tumours by their gene expression profiles. Twenty proteins in the 2-DE maps of breast, lung and ovarian tumours were identified using mass spectrometry. Some of these proteins were found to consist of polypeptide fragments, suggesting the occurrence of proteolytic processing of polypeptides in these tumours. The process of tumour progression leads to the development of tumour heterogeneity. The 2-DE technique was used to study intra- and intertumour heterogeneity. Our results suggested that the degree of intertumour heterogeneity was substantial, whereas intratumoural variations were less pronounced. We conclude that 2-DE separation of proteins in human tumours can yield new information relevant to the understanding of tumour biology. We believe that this line of research will lead to improved diagnostic and predictive tools

Crosstalk between aryl hydrocarbon receptor (AhR) and BCL-2 pathways suggests the use of AhR antagonist to maintain normal differentiation state of mammary epithelial cells during BCL-2 inhibition therapy

IntroductionActivating the aryl hydrocarbon receptor upon exposure to environmental pollutants promotes development of breast cancer stem cell (CSCs). BCL-2 family proteins protect cancer cells from the apoptotic effects of chemotherapeutic drugs. However, the crosstalk between AhR and the BCL-2 family in CSC development remains uninvestigated. ObjectivesThis study explored the interaction mechanisms between AhR and BCL-2 in CSC development and chemoresistance. MethodsA quantitative proteomic analysis study was performed as a tool for comparative expression analysis of breast cancer cells treated by AhR agonist. The basal and inducible levels of BCL-2, AhR, and CYP1A1 in vitro breast cancer and epithelial cell lines and in vivo mice animal models were determined by RT-PCR, Western blot analysis, immunofluorescence, flow cytometry, silencing of the target, and immunohistochemistry. In addition, an in silico toxicity study was conducted using DEREK software. ResultsActivation of the AhR/CYP1A1 pathway in mice increased EpCAMHigh/CD49fLow CD61+ luminal progenitor-like cells in early tumor formation but not in advanced tumors. In parallel, a chemoproteomic study on breast cancer MCF-7 cells revealed that the BCL-2 protein expression was the most upregulated upon AhR activation. The crosstalk between the AhR and BCL-2 pathways in vitro and in vivo modulated the CSCs features and chemoresistance. Interestingly, inhibition of BCL-2 in mice by venetoclax (VCX) increased EpCAMHigh/CD49fLow CD61+ luminal progenitor-like cells, causing inhibition of epithelial lineage markers, disruption of mammary gland branching and induced the epithelial-mesenchymal transition in mammary epithelial cells (MECs). The combined treatment of VCX and AhR antagonists in mice corrected the abnormal differentiation in MECs and protected mammary gland branching and cell identity. ConclusionsThis is the first study to report crosstalk between AhR and BCL-2 in breast CSCs and provides the rationale for using a combined treatment of BCL-2 inhibitor and AhR antagonist for more effective cancer prevention and treatment.- King Faisal Specialist Hospital and Research Centre - grant No. RAC 2130040 - Qatar University - IRCC-2022-48

Crosstalk between aryl hydrocarbon receptor (AhR) and BCL-2 pathways suggests the use of AhR antagonist to maintain normal differentiation state of mammary epithelial cells during BCL-2 inhibition therapy

Introduction: Activating the aryl hydrocarbon receptor upon exposure to environmental pollutants promotes development of breast cancer stem cell (CSCs). BCL-2 family proteins protect cancer cells from the apoptotic effects of chemotherapeutic drugs. However, the crosstalk between AhR and the BCL-2 family in CSC development remains uninvestigated. Objectives: This study explored the interaction mechanisms between AhR and BCL-2 in CSC development and chemoresistance. Methods: A quantitative proteomic analysis study was performed as a tool for comparative expression analysis of breast cancer cells treated by AhR agonist. The basal and inducible levels of BCL-2, AhR, and CYP1A1 in vitro breast cancer and epithelial cell lines and in vivo mice animal models were determined by RT-PCR, Western blot analysis, immunofluorescence, flow cytometry, silencing of the target, and immunohistochemistry. In addition, an in silico toxicity study was conducted using DEREK software. Results: Activation of the AhR/CYP1A1 pathway in mice increased EpCAMHigh/CD49fLow CD61+ luminal progenitor-like cells in early tumor formation but not in advanced tumors. In parallel, a chemoproteomic study on breast cancer MCF-7 cells revealed that the BCL-2 protein expression was the most upregulated upon AhR activation. The crosstalk between the AhR and BCL-2 pathways in vitro and in vivo modulated the CSCs features and chemoresistance. Interestingly, inhibition of BCL-2 in mice by venetoclax (VCX) increased EpCAMHigh/CD49fLow CD61+ luminal progenitor-like cells, causing inhibition of epithelial lineage markers, disruption of mammary gland branching and induced the epithelial-mesenchymal transition in mammary epithelial cells (MECs). The combined treatment of VCX and AhR antagonists in mice corrected the abnormal differentiation in MECs and protected mammary gland branching and cell identity. Conclusions: This is the first study to report crosstalk between AhR and BCL-2 in breast CSCs and provides the rationale for using a combined treatment of BCL-2 inhibitor and AhR antagonist for more effective cancer prevention and treatment

Functional interaction network of 16 genes.

<p>Genes were clustered according to their associated pathways, which are shaded with a different color. Green nodes indicate down-regulated, red, up-regulated, and linker genes (non-colored nodes). The edges represent interactions between genes, with arrows indicating directed interactions and dotted lines indicating predicted relationships.</p

Overlapping KEGG pathways for mRNA and proteomics in in dilated cardiomyopathy.

<p>Overlapping KEGG pathways for mRNA and proteomics in in dilated cardiomyopathy.</p

Integrated Left Ventricular Global Transcriptome and Proteome Profiling in Human End-Stage Dilated Cardiomyopathy - Fig 7

<p>Venn diagrams representing overlap of (A) predicted upstream regulators, (B) enriched GO biological processes, and (C) KEGG pathways between differentially expressed genes and proteins.</p

GO Biological Process and pathway analyses of differentially expressed genes (DEGs) and proteins (DEPs) using the PANTHER classification system.

<p>(A-B) Pie charts displaying significantly enriched biological processes respectively, and (C-D) signaling pathways associated with DEGs and DEPs.</p