title: NFATc1 Links EGFR Signaling to Induction of Sox9 Transcription and Acinar-Ductal Transdifferentiation in the Pancreas.
abstract: OBJECTIVE
  Oncogenic mutations in KRAS contribute to the development of pancreatic ductal adenocarcinoma, but are not sufficient to initiate carcinogenesis. Secondary events, such as inflammation-induced signaling via the epidermal growth factor receptor (EGFR) and expression of the SOX9 gene, are required for tumor formation. Herein we sought to identify the mechanisms that link EGFR signaling with activation of SOX9 during acinar-ductal metaplasia, a transdifferentiation process that precedes pancreatic carcinogenesis.
  
  
  METHODS
  We analyzed pancreatic tissues from Kras(G12D);pdx1-Cre and Kras(G12D);NFATc1(Δ/Δ);pdx1-Cre mice after intraperitoneal administration of caerulein, vs cyclosporin A or dimethyl sulfoxide (controls). Induction of EGFR signaling and its effects on the expression of Nuclear factor of activated T cells c1 (NFATc1) or SOX9 were investigated by quantitative reverse-transcription polymerase chain reaction, immunoblot, and immunohistochemical analyses of mouse and human tissues and acinar cell explants. Interactions between NFATc1 and partner proteins and effects on DNA binding or chromatin modifications were studied using co-immunoprecipitation and chromatin immunoprecipitation assays in acinar cell explants and mouse tissue.
  
  
  RESULTS
  EGFR activation induced expression of NFATc1 in metaplastic areas from patients with chronic pancreatitis and in pancreatic tissue from Kras(G12D) mice. EGFR signaling also promoted formation of a complex between NFATc1 and C-JUN in dedifferentiating mouse acinar cells, leading to activation of Sox9 transcription and induction of acinar-ductal metaplasia. Pharmacologic inhibition of NFATc1 or disruption of the Nfatc1 gene inhibited EGFR-mediated induction of Sox9 transcription and blocked acinar-ductal transdifferentiation and pancreatic cancer initiation in mice.
  
  
  CONCLUSIONS
  EGFR signaling induces expression of NFATc1 and Sox9, leading to acinar cell transdifferentiation and initiation of pancreatic cancer. Strategies designed to disrupt this pathway might be developed to prevent pancreatic cancer initiation in high-risk patients with chronic pancreatitis.
authors.affiliation: Gastroenterology and Gastrointestinal Oncology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Nai-Ming
authors.initials: NM
authors.lastname: Chen
authors.affiliation: Signaling and Transcription Laboratory, Department of Gastroenterology, Philipps-University, Marburg, Germany.
authors.forename: Garima
authors.initials: G
authors.lastname: Singh
authors.affiliation: Gastroenterology and Gastrointestinal Oncology, University Medical Center Goettingen, Goettingen, Germany; Schulze Center for Novel Therapeutics, Division of Oncology Research, Mayo Clinic, Rochester, Minnesota.
authors.forename: Alexander
authors.initials: A
authors.lastname: Koenig
authors.affiliation: Department of Cancer Biology, Mayo Clinic, Jacksonville, Florida.
authors.forename: Geou-Yarh
authors.initials: GY
authors.lastname: Liou
authors.affiliation: Department of Cancer Biology, Mayo Clinic, Jacksonville, Florida.
authors.forename: Peter
authors.initials: P
authors.lastname: Storz
authors.affiliation: Schulze Center for Novel Therapeutics, Division of Oncology Research, Mayo Clinic, Rochester, Minnesota; School of Pharmaceutical Sciences and Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, Zhejiang, People's Republic of China.
authors.forename: Jin-San
authors.initials: JS
authors.lastname: Zhang
authors.affiliation: Signaling and Transcription Laboratory, Department of Gastroenterology, Philipps-University, Marburg, Germany.
authors.forename: Lisanne
authors.initials: L
authors.lastname: Regul
authors.affiliation: Clinic for General, Visceral and Pediatric Surgery, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Sankari
authors.initials: S
authors.lastname: Nagarajan
authors.affiliation: Signaling and Transcription Laboratory, Department of Gastroenterology, Philipps-University, Marburg, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Kühnemuth
authors.affiliation: Clinic for General, Visceral and Pediatric Surgery, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Johnsen
authors.affiliation: Diabetes Center, University of California San Francisco, San Francisco, California.
authors.forename: Matthias
authors.initials: M
authors.lastname: Hebrok
authors.affiliation: II Medizinische Klinik, Klinikum Rechts der Isar, Technische Universität, Munich, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Siveke
authors.affiliation: Schulze Center for Novel Therapeutics, Division of Oncology Research, Mayo Clinic, Rochester, Minnesota.
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Billadeau
authors.affiliation: Gastroenterology and Gastrointestinal Oncology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Ellenrieder
authors.affiliation: Gastroenterology and Gastrointestinal Oncology, University Medical Center Goettingen, Goettingen, Germany. Electronic address: elisabeth.hessmann@med.uni-goettingen.de.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Hessmann
mesh_terms: D000818:Animals; D021441:Carcinoma, Pancreatic Ductal / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D054338:Cell Transdifferentiation; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002108:Ceruletide; D016572:Cyclosporine; D004195:Disease Models, Animal; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D008679:Metaplasia; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation; D050778:NFATC Transcription Factors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D046790:Pancreas, Exocrine / Q000378:metabolism* / Q000473:pathology; D010183:Pancreatic Ducts / Q000378:metabolism* / Q000473:pathology; D010190:Pancreatic Neoplasms / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D010195:Pancreatitis / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D011230:Precancerous Conditions / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D046509:Tissue Culture Techniques; D015533:Transcriptional Activation
chemical_list: D050778:NFATC Transcription Factors; C494003:NFATC1 protein, human; C494005:Nfatc1 protein, mouse; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; C527933:Sox9 protein, mouse; D016572:Cyclosporine; D002108:Ceruletide; C512478:EGFR protein, human; C541485:EGFR protein, mouse; D066246:ErbB Receptors; C495971:Hras protein, mouse; D016283:Proto-Oncogene Proteins p21(ras)
keywords: ChIP; Gene Regulation; Mouse Model; Signal Transduction