387c7808e748ce5b8ebf65f183b2c286 pmc_34944824.pdf 90bd7030ce8aa82653a838e5028c3e5a6e36bc8f pmc_34944824.pdf 5eed7b8563cc8e6e9683dab71d06ba3f40340862f5e1a9205d159171e4b7203d pmc_34944824.pdf Title: TGF Drives Metabolic Perturbations during Epithelial Mesenchymal Transition in Pancreatic Cancer: TGF Induced EMT in PDAC Subject: Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy wherein a majority of patients present metastatic disease at diagnosis. Although the role of epithelial to mesenchymal transition (EMT), mediated by transforming growth factor beta (TGF), in imparting an aggressive phenotype to PDAC is well documented, the underlying biochemical pathway perturbations driving this behaviour have not been elucidated. We used high-resolution mass spectrometry (HRMS) based molecular phenotyping approach in order to delineate metabolic changes concomitant to TGF-induced EMT in pancreatic cancer cells. Strikingly, we observed robust changes in amino acid and energy metabolism that may contribute to tumor invasion and metastasis. Somewhat unexpectedly, TGF treatment resulted in an increase in intracellular levels of retinoic acid (RA) that in turn resulted in increased levels of extracellular matrix (ECM) proteins including fibronectin (FN) and collagen (COL1). These findings were further validated in plasma samples obtained from patients with resectable pancreatic cancer. Taken together, these observations provide novel insights into small molecule dysregulation that triggers a molecular cascade resulting in increased EMT-like changes in pancreatic cancer cells, a paradigm that can be potentially targeted for better clinical outcomes. Keywords: tumor microenvironment; pancreatic cancer; TGF beta; 9-cis retinoic acidPANC-1 cells; epithelial mesenchymal transition Author: Meena U. Rajagopal, Shivani Bansal, Prabhjit Kaur, Shreyans K. Jain, Tatiana Altadil, Charles P. Hinzman, Yaoxiang Li, Joanna Moulton, Baldev Singh, Sunil Bansal, Siddheshwar Kisan Chauthe, Rajbir Singh, Partha P. Banerjee, Mark Mapstone, Massimo S. Fiandaca, Howard J. Federoff, Keith Unger, Jill P. Smith and Amrita K. 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Identity-H yes yes yes 201 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 207 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-03-22 02:26:19 CET RepresentationInformation: pmc_34944824.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-01-10 07:57:28 CET Size: 3306189 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 455 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Children: Item: Title: Cell Lines and Culture Conditions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: UPLC-ESI-QTOF MS Analysis Destination: 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Children: Item: Title: MRM-MS Quantitation of Intra-Cellular Levels of Retinoic Acid and Amino Acids in PANC-1 Cells Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: MRM-MS Based Quantitation of Metabolites (Acetyl CoA) in Human Plasma Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Enzyme Linked Immuno Sorbent Assay (ELISA) of Fibronectin (FN1) and O-Linked N-Acetylglucosamine Transferase (OGT) in Human Plasma Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Children: Item: Title: TGF Induced EMT in PANC-1 Cells Is Accompanied by Robust Changes in Metabolic Profiles Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: TGF and 9-cis RA Treatment Markedly Dysregulates Genes Involved in ECM Remodeling in PANC-1 Cells Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: TGF and 9-cis RA Promote EMT Induced O-Glycosylation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Plasma Fibronectin, OGT and Acetyl CoA Levels Are Significantly Increased in Patients Diagnosed with Early Stage Pancreatic Cancer Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Info: Title: TGF Drives Metabolic Perturbations during Epithelial Mesenchymal Transition in Pancreatic Cancer: TGF Induced EMT in PDAC Author: Meena U. Rajagopal, Shivani Bansal, Prabhjit Kaur, Shreyans K. Jain, Tatiana Altadil, Charles P. Hinzman, Yaoxiang Li, Joanna Moulton, Baldev Singh, Sunil Bansal, Siddheshwar Kisan Chauthe, Rajbir Singh, Partha P. Banerjee, Mark Mapstone, Massimo S. Fiandaca, Howard J. Federoff, Keith Unger, Jill P. Smith and Amrita K. Cheema Subject: Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy wherein a majority of patients present metastatic disease at diagnosis. Although the role of epithelial to mesenchymal transition (EMT), mediated by transforming growth factor beta (TGF), in imparting an aggressive phenotype to PDAC is well documented, the underlying biochemical pathway perturbations driving this behaviour have not been elucidated. We used high-resolution mass spectrometry (HRMS) based molecular phenotyping approach in order to delineate metabolic changes concomitant to TGF-induced EMT in pancreatic cancer cells. Strikingly, we observed robust changes in amino acid and energy metabolism that may contribute to tumor invasion and metastasis. Somewhat unexpectedly, TGF treatment resulted in an increase in intracellular levels of retinoic acid (RA) that in turn resulted in increased levels of extracellular matrix (ECM) proteins including fibronectin (FN) and collagen (COL1). These findings were further validated in plasma samples obtained from patients with resectable pancreatic cancer. Taken together, these observations provide novel insights into small molecule dysregulation that triggers a molecular cascade resulting in increased EMT-like changes in pancreatic cancer cells, a paradigm that can be potentially targeted for better clinical outcomes. 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