NFATc1 links EGFR signaling to induction of sox9 transcription and acinar-ductal transdifferentiation in the pancreas

Nai Ming Chen, Garima Singh, Alexander Koenig, Geou Yarh Liou, Peter Storz, Jin San Zhang, Lisanne Regul, Sankari Nagarajan, Benjamin Kühnemuth, Steven Johnsen, Matthias Hebrok, Jens Siveke, Daniel D Billadeau, Volker Ellenrieder, Elisabeth Hessmann

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Background & Aims 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<sup>G12D</sup>;pdx1-Cre and Kras<sup>G12D</sup>;NFATc1<sup>Δ/Δ</sup>;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<sup>G12D</sup> 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.

Original languageEnglish (US)
Pages (from-to)1024-1034.e9
JournalGastroenterology
Volume148
Issue number5
DOIs
StatePublished - May 1 2015

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NFATC Transcription Factors
Epidermal Growth Factor Receptor
Pancreas
Acinar Cells
Pancreatic Neoplasms
Chronic Pancreatitis
Metaplasia
Carcinogenesis
Cell Transdifferentiation
erbB-1 Genes
Ceruletide
Chromatin Immunoprecipitation
Dimethyl Sulfoxide
Immunoprecipitation
Transcriptional Activation
Cyclosporine
Reverse Transcription
Chromatin
Adenocarcinoma
Inflammation

Keywords

  • ChIP
  • Gene Regulation
  • Mouse Model
  • Signal Transduction

ASJC Scopus subject areas

  • Gastroenterology

Cite this

NFATc1 links EGFR signaling to induction of sox9 transcription and acinar-ductal transdifferentiation in the pancreas. / Chen, Nai Ming; Singh, Garima; Koenig, Alexander; Liou, Geou Yarh; Storz, Peter; Zhang, Jin San; Regul, Lisanne; Nagarajan, Sankari; Kühnemuth, Benjamin; Johnsen, Steven; Hebrok, Matthias; Siveke, Jens; Billadeau, Daniel D; Ellenrieder, Volker; Hessmann, Elisabeth.

In: Gastroenterology, Vol. 148, No. 5, 01.05.2015, p. 1024-1034.e9.

Research output: Contribution to journalArticle

Chen, NM, Singh, G, Koenig, A, Liou, GY, Storz, P, Zhang, JS, Regul, L, Nagarajan, S, Kühnemuth, B, Johnsen, S, Hebrok, M, Siveke, J, Billadeau, DD, Ellenrieder, V & Hessmann, E 2015, 'NFATc1 links EGFR signaling to induction of sox9 transcription and acinar-ductal transdifferentiation in the pancreas', Gastroenterology, vol. 148, no. 5, pp. 1024-1034.e9. https://doi.org/10.1053/j.gastro.2015.01.033
Chen, Nai Ming ; Singh, Garima ; Koenig, Alexander ; Liou, Geou Yarh ; Storz, Peter ; Zhang, Jin San ; Regul, Lisanne ; Nagarajan, Sankari ; Kühnemuth, Benjamin ; Johnsen, Steven ; Hebrok, Matthias ; Siveke, Jens ; Billadeau, Daniel D ; Ellenrieder, Volker ; Hessmann, Elisabeth. / NFATc1 links EGFR signaling to induction of sox9 transcription and acinar-ductal transdifferentiation in the pancreas. In: Gastroenterology. 2015 ; Vol. 148, No. 5. pp. 1024-1034.e9.
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abstract = "Background & Aims 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 KrasG12D;pdx1-Cre and KrasG12D;NFATc1Δ/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 KrasG12D 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.",
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T1 - NFATc1 links EGFR signaling to induction of sox9 transcription and acinar-ductal transdifferentiation in the pancreas

AU - Chen, Nai Ming

AU - Singh, Garima

AU - Koenig, Alexander

AU - Liou, Geou Yarh

AU - Storz, Peter

AU - Zhang, Jin San

AU - Regul, Lisanne

AU - Nagarajan, Sankari

AU - Kühnemuth, Benjamin

AU - Johnsen, Steven

AU - Hebrok, Matthias

AU - Siveke, Jens

AU - Billadeau, Daniel D

AU - Ellenrieder, Volker

AU - Hessmann, Elisabeth

PY - 2015/5/1

Y1 - 2015/5/1

N2 - Background & Aims 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 KrasG12D;pdx1-Cre and KrasG12D;NFATc1Δ/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 KrasG12D 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.

AB - Background & Aims 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 KrasG12D;pdx1-Cre and KrasG12D;NFATc1Δ/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 KrasG12D 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.

KW - ChIP

KW - Gene Regulation

KW - Mouse Model

KW - Signal Transduction

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