Journal Article

Protein Tyrosine Phosphatase Nonreceptor Type 2 Regulates Autophagosome Formation in Human Intestinal Cells

Rogler, Gerhard, Vavricka, Stephan R., Fischbeck, Anne, Fried, Michael, Wojtal, Kacper A., Scharl, Michael, Frei, Pascal, McCole, Declan F., Becker, Helen M., Arikkat, Joba, Weber, Achim, Kellermeier, Silvia, Boone, David L., Pesch, Theresa, Loessner, Martin J.

2012

Overview

Autophagy is a process of central importance for maintaining cell homeostasis, survival, and the regulation of inflammation. Recent studies associated variants within the gene loci, encoding protein tyrosine phosphatase nonreceptor type 2 (PTPN2), and autophagy genes, such as autophagy-related 16-like 1 (ATG16L1), with chronic inflammatory disorders, such as Crohn's disease (CD). We show that PTPN2 regulates autophagy in human intestinal epithelial cells (IEC) and primary colonic lamina propria fibroblasts (CLPF).MethodsProtein analysis in IEC and CLPF was performed by western blotting. Autophagososme formation was assessed by LC3B immunofluorescence or immunohistochemistry. Human intestinal tissue samples were obtained from noninflammatory bowel disease (IBD) control or from CD patients and genotyped for disease-associated PTPN2 or ATG16L1 variations.ResultsKnockdown of PTPN2 causes impaired autophagosome formation and dysfunctional autophagy resulted in increased levels of intracellular Listeria monocytogenes (LM) and elevated IEC apoptosis in response to tumor necrosis factor (TNF) and interferon gamma (IFN-γ). Similar findings were observed in primary CLPF derived from CD patients carrying the CD-associated PTPN2 variant. Presence of the ATG16L1 variant prevented the cytokine-induced rise in PTPN2 protein, finally resulting in impaired LC3B-II levels in IEC. Actively inflamed intestinal biopsies from CD patients carrying either ATG16L1 or PTPN2 genetic variants revealed aberrant LC3B expression patterns when compared with samples from non-IBD control patients.ConclusionsOur results demonstrate that PTPN2 regulates autophagosome formation in human intestinal cells. We provide a model of how a dysfunction of the CD susceptibility genes, PTPN2 and/or ATG16L1, may contribute to the onset and perpetuation of chronic intestinal inflammation.

• Publisher: Oxford University Press,Wiley Subscription Services, Inc., A Wiley Company
• Subject: Adult / Aged / ATG16L1 / Autophagy / Autophagy-Related Proteins / Carrier Proteins - metabolism / Case-Control Studies / Cell Communication / Cells, Cultured / Colon - metabolism / Colon - pathology / Crohn Disease - metabolism / Crohn Disease - microbiology / Crohn Disease - pathology / Crohn's disease / Cytokines - metabolism / Female / Fibroblasts - metabolism / Fibroblasts - pathology / Humans / Immunoenzyme Techniques / Intestines - metabolism / Intestines - pathology / Listeria monocytogenes - pathogenicity / Listeriosis - metabolism / Listeriosis - microbiology / Listeriosis - pathology / Male / Microscopy, Fluorescence / Middle Aged / Mucous Membrane - metabolism / Mucous Membrane - pathology / Phosphatase / Phosphorylation / Prognosis / Prospective Studies / Protein Tyrosine Phosphatase, Non-Receptor Type 2 - antagonists & inhibitors / Protein Tyrosine Phosphatase, Non-Receptor Type 2 - genetics / Protein Tyrosine Phosphatase, Non-Receptor Type 2 - metabolism / Proteins / PTPN2 / RNA, Small Interfering - genetics / TOR Serine-Threonine Kinases - metabolism / Tumor necrosis factor-TNF