ETS-Transcription Factor ETV1 Regulates Stromal Expansion and Metastasis in Pancreatic Cancer

Steffen Heeg, Koushik K. Das, Maximilian Reichert, Basil Bakir, Shigetsugu Takano, Julia Caspers, Nicole M. Aiello, Katherine Wu, Albrecht Neesse, Anirban Maitra, Christine A. Iacobuzio-Donahue, Philip Hicks, Anil K. Rustgi

Research output: Contribution to journalArticle

Abstract

Background & Aims The ETS-transcription factor ETV1 is involved in epithelial−mesenchymal transition during pancreatic development and is induced in mouse pancreatic intraepithelial neoplasia (PanIN) and pancreatic ductal adenocarcinoma (PDAC). We investigated the function of ETV1 in stromal expansion of PDAC and metastasis, as well as its effects on a novel downstream target Sparc, which encodes a matricellular protein found in PDAC stroma that has been associated with invasiveness, metastasis and poor patient outcomes. Methods Pancreatic ductal cells were isolated from Pdx1Cre;KrasG12D/+ mice (PanIN), Pdx1Cre;KrasG12D/+;p53fl/+ and Pdx1Cre;KrasG12D/+;p53fl/+;Rosa26YFP mice (PDAC), and Pdx1Cre;KrasG12D/+;p53fl/+;Sparc−/− mice. Cells were grown in 3-dimensional organoid culture to analyze morphology, proliferation, and invasion. Human PanIN and PDAC tissues were evaluated for ETV1 expression. Orthotopic pancreatic transplants of ETV1-overexpressing PDAC and respective control cells were performed. Results ETV1 expression was significantly increased in human PanINs and, even more so, in primary and metastatic PDAC. Analyses of mouse orthotopic xenografts revealed that ETV1 induced significantly larger primary tumors than controls, with significantly increased stromal expansion, ascites and metastases. In 3-dimensional organoids, ETV1 disrupted cyst architecture, induced EMT, and increased invasive capacity. Furthermore, we identified Sparc as a novel functional gene target of Etv1 by luciferase assays, and SPARC and ETV1 proteins co-localized in vivo. Disruption of Sparc abrogates the phenotype of stromal expansion and metastasis found with ETV1 overexpression in vivo. We identified hyaluronan synthase 2 (Has2) as another novel downstream factor of Etv1; that may mediate ETV1's significant expansion of hyaluronic acid in PDAC stroma. Conversely, disruption of Etv1 in PDAC mice (Pdx1Cre;KrasG12D/+;p53fl/+;Rosa26YFP;Cre;Etv1fl/fl) reduced levels of SPARC and hyaluronic acid in the stroma. Conclusions ETV1 is critical in the desmoplastic stromal expansion and metastatic progression of pancreatic cancer in mice, mediated functionally in part through Sparc and Has2.

Original languageEnglish (US)
Pages (from-to)540-553.e14
JournalGastroenterology
Volume151
Issue number3
DOIs
StatePublished - Sep 1 2016
Externally publishedYes

Fingerprint

Pancreatic Neoplasms
Adenocarcinoma
Transcription Factors
Organoids
Neoplasm Metastasis
Hyaluronic Acid
Neoplasms
Luciferases
Ascites
Heterografts
Cysts
Proteins
Transplants
Phenotype
Genes

Keywords

  • Cancer
  • EMT
  • Gene Regulation
  • Pancreas

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Heeg, S., Das, K. K., Reichert, M., Bakir, B., Takano, S., Caspers, J., ... Rustgi, A. K. (2016). ETS-Transcription Factor ETV1 Regulates Stromal Expansion and Metastasis in Pancreatic Cancer. Gastroenterology, 151(3), 540-553.e14. https://doi.org/10.1053/j.gastro.2016.06.005

ETS-Transcription Factor ETV1 Regulates Stromal Expansion and Metastasis in Pancreatic Cancer. / Heeg, Steffen; Das, Koushik K.; Reichert, Maximilian; Bakir, Basil; Takano, Shigetsugu; Caspers, Julia; Aiello, Nicole M.; Wu, Katherine; Neesse, Albrecht; Maitra, Anirban; Iacobuzio-Donahue, Christine A.; Hicks, Philip; Rustgi, Anil K.

In: Gastroenterology, Vol. 151, No. 3, 01.09.2016, p. 540-553.e14.

Research output: Contribution to journalArticle

Heeg, S, Das, KK, Reichert, M, Bakir, B, Takano, S, Caspers, J, Aiello, NM, Wu, K, Neesse, A, Maitra, A, Iacobuzio-Donahue, CA, Hicks, P & Rustgi, AK 2016, 'ETS-Transcription Factor ETV1 Regulates Stromal Expansion and Metastasis in Pancreatic Cancer', Gastroenterology, vol. 151, no. 3, pp. 540-553.e14. https://doi.org/10.1053/j.gastro.2016.06.005
Heeg, Steffen ; Das, Koushik K. ; Reichert, Maximilian ; Bakir, Basil ; Takano, Shigetsugu ; Caspers, Julia ; Aiello, Nicole M. ; Wu, Katherine ; Neesse, Albrecht ; Maitra, Anirban ; Iacobuzio-Donahue, Christine A. ; Hicks, Philip ; Rustgi, Anil K. / ETS-Transcription Factor ETV1 Regulates Stromal Expansion and Metastasis in Pancreatic Cancer. In: Gastroenterology. 2016 ; Vol. 151, No. 3. pp. 540-553.e14.
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abstract = "Background & Aims The ETS-transcription factor ETV1 is involved in epithelial−mesenchymal transition during pancreatic development and is induced in mouse pancreatic intraepithelial neoplasia (PanIN) and pancreatic ductal adenocarcinoma (PDAC). We investigated the function of ETV1 in stromal expansion of PDAC and metastasis, as well as its effects on a novel downstream target Sparc, which encodes a matricellular protein found in PDAC stroma that has been associated with invasiveness, metastasis and poor patient outcomes. Methods Pancreatic ductal cells were isolated from Pdx1Cre;KrasG12D/+ mice (PanIN), Pdx1Cre;KrasG12D/+;p53fl/+ and Pdx1Cre;KrasG12D/+;p53fl/+;Rosa26YFP mice (PDAC), and Pdx1Cre;KrasG12D/+;p53fl/+;Sparc−/− mice. Cells were grown in 3-dimensional organoid culture to analyze morphology, proliferation, and invasion. Human PanIN and PDAC tissues were evaluated for ETV1 expression. Orthotopic pancreatic transplants of ETV1-overexpressing PDAC and respective control cells were performed. Results ETV1 expression was significantly increased in human PanINs and, even more so, in primary and metastatic PDAC. Analyses of mouse orthotopic xenografts revealed that ETV1 induced significantly larger primary tumors than controls, with significantly increased stromal expansion, ascites and metastases. In 3-dimensional organoids, ETV1 disrupted cyst architecture, induced EMT, and increased invasive capacity. Furthermore, we identified Sparc as a novel functional gene target of Etv1 by luciferase assays, and SPARC and ETV1 proteins co-localized in vivo. Disruption of Sparc abrogates the phenotype of stromal expansion and metastasis found with ETV1 overexpression in vivo. We identified hyaluronan synthase 2 (Has2) as another novel downstream factor of Etv1; that may mediate ETV1's significant expansion of hyaluronic acid in PDAC stroma. Conversely, disruption of Etv1 in PDAC mice (Pdx1Cre;KrasG12D/+;p53fl/+;Rosa26YFP;Cre;Etv1fl/fl) reduced levels of SPARC and hyaluronic acid in the stroma. Conclusions ETV1 is critical in the desmoplastic stromal expansion and metastatic progression of pancreatic cancer in mice, mediated functionally in part through Sparc and Has2.",
keywords = "Cancer, EMT, Gene Regulation, Pancreas",
author = "Steffen Heeg and Das, {Koushik K.} and Maximilian Reichert and Basil Bakir and Shigetsugu Takano and Julia Caspers and Aiello, {Nicole M.} and Katherine Wu and Albrecht Neesse and Anirban Maitra and Iacobuzio-Donahue, {Christine A.} and Philip Hicks and Rustgi, {Anil K.}",
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T1 - ETS-Transcription Factor ETV1 Regulates Stromal Expansion and Metastasis in Pancreatic Cancer

AU - Heeg, Steffen

AU - Das, Koushik K.

AU - Reichert, Maximilian

AU - Bakir, Basil

AU - Takano, Shigetsugu

AU - Caspers, Julia

AU - Aiello, Nicole M.

AU - Wu, Katherine

AU - Neesse, Albrecht

AU - Maitra, Anirban

AU - Iacobuzio-Donahue, Christine A.

AU - Hicks, Philip

AU - Rustgi, Anil K.

PY - 2016/9/1

Y1 - 2016/9/1

N2 - Background & Aims The ETS-transcription factor ETV1 is involved in epithelial−mesenchymal transition during pancreatic development and is induced in mouse pancreatic intraepithelial neoplasia (PanIN) and pancreatic ductal adenocarcinoma (PDAC). We investigated the function of ETV1 in stromal expansion of PDAC and metastasis, as well as its effects on a novel downstream target Sparc, which encodes a matricellular protein found in PDAC stroma that has been associated with invasiveness, metastasis and poor patient outcomes. Methods Pancreatic ductal cells were isolated from Pdx1Cre;KrasG12D/+ mice (PanIN), Pdx1Cre;KrasG12D/+;p53fl/+ and Pdx1Cre;KrasG12D/+;p53fl/+;Rosa26YFP mice (PDAC), and Pdx1Cre;KrasG12D/+;p53fl/+;Sparc−/− mice. Cells were grown in 3-dimensional organoid culture to analyze morphology, proliferation, and invasion. Human PanIN and PDAC tissues were evaluated for ETV1 expression. Orthotopic pancreatic transplants of ETV1-overexpressing PDAC and respective control cells were performed. Results ETV1 expression was significantly increased in human PanINs and, even more so, in primary and metastatic PDAC. Analyses of mouse orthotopic xenografts revealed that ETV1 induced significantly larger primary tumors than controls, with significantly increased stromal expansion, ascites and metastases. In 3-dimensional organoids, ETV1 disrupted cyst architecture, induced EMT, and increased invasive capacity. Furthermore, we identified Sparc as a novel functional gene target of Etv1 by luciferase assays, and SPARC and ETV1 proteins co-localized in vivo. Disruption of Sparc abrogates the phenotype of stromal expansion and metastasis found with ETV1 overexpression in vivo. We identified hyaluronan synthase 2 (Has2) as another novel downstream factor of Etv1; that may mediate ETV1's significant expansion of hyaluronic acid in PDAC stroma. Conversely, disruption of Etv1 in PDAC mice (Pdx1Cre;KrasG12D/+;p53fl/+;Rosa26YFP;Cre;Etv1fl/fl) reduced levels of SPARC and hyaluronic acid in the stroma. Conclusions ETV1 is critical in the desmoplastic stromal expansion and metastatic progression of pancreatic cancer in mice, mediated functionally in part through Sparc and Has2.

AB - Background & Aims The ETS-transcription factor ETV1 is involved in epithelial−mesenchymal transition during pancreatic development and is induced in mouse pancreatic intraepithelial neoplasia (PanIN) and pancreatic ductal adenocarcinoma (PDAC). We investigated the function of ETV1 in stromal expansion of PDAC and metastasis, as well as its effects on a novel downstream target Sparc, which encodes a matricellular protein found in PDAC stroma that has been associated with invasiveness, metastasis and poor patient outcomes. Methods Pancreatic ductal cells were isolated from Pdx1Cre;KrasG12D/+ mice (PanIN), Pdx1Cre;KrasG12D/+;p53fl/+ and Pdx1Cre;KrasG12D/+;p53fl/+;Rosa26YFP mice (PDAC), and Pdx1Cre;KrasG12D/+;p53fl/+;Sparc−/− mice. Cells were grown in 3-dimensional organoid culture to analyze morphology, proliferation, and invasion. Human PanIN and PDAC tissues were evaluated for ETV1 expression. Orthotopic pancreatic transplants of ETV1-overexpressing PDAC and respective control cells were performed. Results ETV1 expression was significantly increased in human PanINs and, even more so, in primary and metastatic PDAC. Analyses of mouse orthotopic xenografts revealed that ETV1 induced significantly larger primary tumors than controls, with significantly increased stromal expansion, ascites and metastases. In 3-dimensional organoids, ETV1 disrupted cyst architecture, induced EMT, and increased invasive capacity. Furthermore, we identified Sparc as a novel functional gene target of Etv1 by luciferase assays, and SPARC and ETV1 proteins co-localized in vivo. Disruption of Sparc abrogates the phenotype of stromal expansion and metastasis found with ETV1 overexpression in vivo. We identified hyaluronan synthase 2 (Has2) as another novel downstream factor of Etv1; that may mediate ETV1's significant expansion of hyaluronic acid in PDAC stroma. Conversely, disruption of Etv1 in PDAC mice (Pdx1Cre;KrasG12D/+;p53fl/+;Rosa26YFP;Cre;Etv1fl/fl) reduced levels of SPARC and hyaluronic acid in the stroma. Conclusions ETV1 is critical in the desmoplastic stromal expansion and metastatic progression of pancreatic cancer in mice, mediated functionally in part through Sparc and Has2.

KW - Cancer

KW - EMT

KW - Gene Regulation

KW - Pancreas

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