title: Protein tyrosine phosphatase nonreceptor type 2 regulates autophagosome formation in human intestinal cells.
abstract: BACKGROUND
  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).
  
  
  METHODS
  Protein 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.
  
  
  RESULTS
  Knockdown 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.
  
  
  CONCLUSIONS
  Our 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.
authors: Division of Gastroenterology and Hepatology, University Hospital Zurich, Zurich, Switzerland. michael.scharl@usz.ch
authors: Michael
authors: M
authors: Scharl
authors: Kacper A
authors: KA
authors: Wojtal
authors: Helen M
authors: HM
authors: Becker
authors: Anne
authors: A
authors: Fischbeck
authors: Pascal
authors: P
authors: Frei
authors: Joba
authors: J
authors: Arikkat
authors: Theresa
authors: T
authors: Pesch
authors: Silvia
authors: S
authors: Kellermeier
authors: David L
authors: DL
authors: Boone
authors: Achim
authors: A
authors: Weber
authors: Martin J
authors: MJ
authors: Loessner
authors: Stephan R
authors: SR
authors: Vavricka
authors: Michael
authors: M
authors: Fried
authors: Declan F
authors: DF
authors: McCole
authors: Gerhard
authors: G
authors: Rogler
mesh_terms: D000328:Adult; D000368:Aged; D001343:Autophagy; D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins / Q000378:metabolism*; D016022:Case-Control Studies; D002450:Cell Communication; D002478:Cells, Cultured; D003106:Colon / Q000378:metabolism / Q000473:pathology; D003424:Crohn Disease / Q000378:metabolism / Q000382:microbiology / Q000473:pathology*; D016207:Cytokines / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D007124:Immunoenzyme Techniques; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000473:pathology*; D008089:Listeria monocytogenes / Q000472:pathogenicity; D008088:Listeriosis / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D009092:Mucous Membrane / Q000378:metabolism / Q000473:pathology; D010766:Phosphorylation; D011379:Prognosis; D011446:Prospective Studies; D054578:Protein Tyrosine Phosphatase, Non-Receptor Type 2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism
chemical_list: C518959:ATG16L1 protein, human; D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins; D016207:Cytokines; D034741:RNA, Small Interfering; D058570:TOR Serine-Threonine Kinases; C516649:PTPN2 protein, human; D054578:Protein Tyrosine Phosphatase, Non-Receptor Type 2