FoxO3 increases miR-34a to cause palmitate-induced cholangiocyte lipoapoptosis

Sathish K Natarajan, Bailey A. Stringham, Ashley M. Mohr, Cody J. Wehrkamp, Sizhao Lu, Mary Anne Phillippi, Dee Harrison-Findik, Justin L Mott

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Nonalcoholic steatohepatitis (NASH) patients have elevated plasma saturated free fatty acid levels. These toxic fatty acids can induce liver cell death and our recent results demonstrated that the biliary epithelium may be susceptible to lipotoxicity. Here, we explored the molecular mechanisms of cholangiocyte lipoapoptosis in cell culture and in an animal model of NASH. Treatment of cholangiocytes with palmitate (PA) showed increased caspase 3/7 activity and increased levels of cleaved poly (ADP-ribose) polymerase and cleaved caspase 3, demonstrating cholangiocyte lipoapoptosis. Interestingly, treatment with PA significantly increased the levels of microRNA miR-34a, a pro-apoptotic microRNA known to be elevated in NASH. PA induction of miR-34a was abolished in cholangiocytes transduced with forkhead family of transcription factor class O (FoxO)3 shRNA, demonstrating that FoxO3 activation is upstream of miR-34a and suggesting that FoxO3 is a novel transcriptional regulator of miR-34a. Further, anti-miR-34a protected cholangiocytes from PA-induced lipoapoptosis. Direct and indirect targets of miR-34a, such as SIRT1, receptor tyrosine kinase (MET), Kruppel-like factor 4, fibroblast growth factor receptor (FGFR)1, and FGFR4, were all decreased in PA-treated cholangiocytes. SIRT1 and MET were partially rescued by a miR-34a antagonist. Cholangiocyte apoptosis and miR-34a were dramatically increased in the liver of mice with early histologic features of NASH. Our study provides evidence for the pro-apoptotic role of miR-34a in PA-induced cholangiocyte lipoapoptosis in culture and in the liver.

Original languageEnglish (US)
Pages (from-to)866-875
Number of pages10
JournalJournal of Lipid Research
Volume58
Issue number5
DOIs
StatePublished - May 2017

Fingerprint

Palmitates
Liver
MicroRNAs
Caspase 3
Fatty Acids
Receptor, Fibroblast Growth Factor, Type 1
Proto-Oncogene Proteins c-met
Forkhead Transcription Factors
Caspase 7
Poly(ADP-ribose) Polymerases
Poisons
Cell death
Cell culture
Nonesterified Fatty Acids
Small Interfering RNA
Animals
Cell Death
Transcription Factors
Epithelium
Animal Models

Keywords

  • Forkhead box transcription factor class O3
  • MicroRNA-34a
  • Nonalcoholic fatty liver disease
  • Steatohepatitis

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

FoxO3 increases miR-34a to cause palmitate-induced cholangiocyte lipoapoptosis. / Natarajan, Sathish K; Stringham, Bailey A.; Mohr, Ashley M.; Wehrkamp, Cody J.; Lu, Sizhao; Phillippi, Mary Anne; Harrison-Findik, Dee; Mott, Justin L.

In: Journal of Lipid Research, Vol. 58, No. 5, 05.2017, p. 866-875.

Research output: Contribution to journalArticle

Natarajan, SK, Stringham, BA, Mohr, AM, Wehrkamp, CJ, Lu, S, Phillippi, MA, Harrison-Findik, D & Mott, JL 2017, 'FoxO3 increases miR-34a to cause palmitate-induced cholangiocyte lipoapoptosis', Journal of Lipid Research, vol. 58, no. 5, pp. 866-875. https://doi.org/10.1194/jlr.M071357
Natarajan, Sathish K ; Stringham, Bailey A. ; Mohr, Ashley M. ; Wehrkamp, Cody J. ; Lu, Sizhao ; Phillippi, Mary Anne ; Harrison-Findik, Dee ; Mott, Justin L. / FoxO3 increases miR-34a to cause palmitate-induced cholangiocyte lipoapoptosis. In: Journal of Lipid Research. 2017 ; Vol. 58, No. 5. pp. 866-875.
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title = "FoxO3 increases miR-34a to cause palmitate-induced cholangiocyte lipoapoptosis",
abstract = "Nonalcoholic steatohepatitis (NASH) patients have elevated plasma saturated free fatty acid levels. These toxic fatty acids can induce liver cell death and our recent results demonstrated that the biliary epithelium may be susceptible to lipotoxicity. Here, we explored the molecular mechanisms of cholangiocyte lipoapoptosis in cell culture and in an animal model of NASH. Treatment of cholangiocytes with palmitate (PA) showed increased caspase 3/7 activity and increased levels of cleaved poly (ADP-ribose) polymerase and cleaved caspase 3, demonstrating cholangiocyte lipoapoptosis. Interestingly, treatment with PA significantly increased the levels of microRNA miR-34a, a pro-apoptotic microRNA known to be elevated in NASH. PA induction of miR-34a was abolished in cholangiocytes transduced with forkhead family of transcription factor class O (FoxO)3 shRNA, demonstrating that FoxO3 activation is upstream of miR-34a and suggesting that FoxO3 is a novel transcriptional regulator of miR-34a. Further, anti-miR-34a protected cholangiocytes from PA-induced lipoapoptosis. Direct and indirect targets of miR-34a, such as SIRT1, receptor tyrosine kinase (MET), Kruppel-like factor 4, fibroblast growth factor receptor (FGFR)1, and FGFR4, were all decreased in PA-treated cholangiocytes. SIRT1 and MET were partially rescued by a miR-34a antagonist. Cholangiocyte apoptosis and miR-34a were dramatically increased in the liver of mice with early histologic features of NASH. Our study provides evidence for the pro-apoptotic role of miR-34a in PA-induced cholangiocyte lipoapoptosis in culture and in the liver.",
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author = "Natarajan, {Sathish K} and Stringham, {Bailey A.} and Mohr, {Ashley M.} and Wehrkamp, {Cody J.} and Sizhao Lu and Phillippi, {Mary Anne} and Dee Harrison-Findik and Mott, {Justin L}",
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AU - Natarajan, Sathish K

AU - Stringham, Bailey A.

AU - Mohr, Ashley M.

AU - Wehrkamp, Cody J.

AU - Lu, Sizhao

AU - Phillippi, Mary Anne

AU - Harrison-Findik, Dee

AU - Mott, Justin L

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N2 - Nonalcoholic steatohepatitis (NASH) patients have elevated plasma saturated free fatty acid levels. These toxic fatty acids can induce liver cell death and our recent results demonstrated that the biliary epithelium may be susceptible to lipotoxicity. Here, we explored the molecular mechanisms of cholangiocyte lipoapoptosis in cell culture and in an animal model of NASH. Treatment of cholangiocytes with palmitate (PA) showed increased caspase 3/7 activity and increased levels of cleaved poly (ADP-ribose) polymerase and cleaved caspase 3, demonstrating cholangiocyte lipoapoptosis. Interestingly, treatment with PA significantly increased the levels of microRNA miR-34a, a pro-apoptotic microRNA known to be elevated in NASH. PA induction of miR-34a was abolished in cholangiocytes transduced with forkhead family of transcription factor class O (FoxO)3 shRNA, demonstrating that FoxO3 activation is upstream of miR-34a and suggesting that FoxO3 is a novel transcriptional regulator of miR-34a. Further, anti-miR-34a protected cholangiocytes from PA-induced lipoapoptosis. Direct and indirect targets of miR-34a, such as SIRT1, receptor tyrosine kinase (MET), Kruppel-like factor 4, fibroblast growth factor receptor (FGFR)1, and FGFR4, were all decreased in PA-treated cholangiocytes. SIRT1 and MET were partially rescued by a miR-34a antagonist. Cholangiocyte apoptosis and miR-34a were dramatically increased in the liver of mice with early histologic features of NASH. Our study provides evidence for the pro-apoptotic role of miR-34a in PA-induced cholangiocyte lipoapoptosis in culture and in the liver.

AB - Nonalcoholic steatohepatitis (NASH) patients have elevated plasma saturated free fatty acid levels. These toxic fatty acids can induce liver cell death and our recent results demonstrated that the biliary epithelium may be susceptible to lipotoxicity. Here, we explored the molecular mechanisms of cholangiocyte lipoapoptosis in cell culture and in an animal model of NASH. Treatment of cholangiocytes with palmitate (PA) showed increased caspase 3/7 activity and increased levels of cleaved poly (ADP-ribose) polymerase and cleaved caspase 3, demonstrating cholangiocyte lipoapoptosis. Interestingly, treatment with PA significantly increased the levels of microRNA miR-34a, a pro-apoptotic microRNA known to be elevated in NASH. PA induction of miR-34a was abolished in cholangiocytes transduced with forkhead family of transcription factor class O (FoxO)3 shRNA, demonstrating that FoxO3 activation is upstream of miR-34a and suggesting that FoxO3 is a novel transcriptional regulator of miR-34a. Further, anti-miR-34a protected cholangiocytes from PA-induced lipoapoptosis. Direct and indirect targets of miR-34a, such as SIRT1, receptor tyrosine kinase (MET), Kruppel-like factor 4, fibroblast growth factor receptor (FGFR)1, and FGFR4, were all decreased in PA-treated cholangiocytes. SIRT1 and MET were partially rescued by a miR-34a antagonist. Cholangiocyte apoptosis and miR-34a were dramatically increased in the liver of mice with early histologic features of NASH. Our study provides evidence for the pro-apoptotic role of miR-34a in PA-induced cholangiocyte lipoapoptosis in culture and in the liver.

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KW - Nonalcoholic fatty liver disease

KW - Steatohepatitis

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