Hyperinsulinemic hypoglycemia subtype glucokinase V91L mutant induces necrosis in β-cells via ATP depletion

Brian Lu, Jason M. Tonne, Miguel Munoz-Gomez, Yasuhiro H Ikeda

Research output: Contribution to journalArticle

Abstract

Hyperinsulinemic hypoglycemia subtype glucokinase (GCK-HH) is caused by an activating mutation in glucokinase (GCK) and has been shown to increase β-cell death. However, the mechanism of β-cell death in GCK-HH remains poorly understood. Here, we expressed the GCK-HH V91L GCK mutant in INS-1 832/13 cells to determine the effect of the mutation on β-cell viability and the mechanisms of β-cell death. We showed that expression of the V91L GCK mutant in INS-1 832/13 cells resulted in a rapid glucose concentration-dependent loss of cell viability. At 11 mM D-glucose, INS-1 832/13 cells expressing V91L GCK showed increased cell permeability without significant increases in Annexin V staining or caspase 3/7 activation, indicating that these cells are primarily undergoing cell death via necrosis. Over-expression of SV40 large T antigen, which inhibits the p53 pathway, did not affect the V91L GCK-induced cell death. We also found that non-phosphorylatable L-glucose did not induce rapid cell death. Of note, glucose phosphorylation coincided with a 90% loss of intracellular ATP content. Thus, our data suggest that the GCK V91L mutant induces rapid necrosis in INS-1 cells through accelerated glucose phosphorylation, ATP depletion, and increased cell permeability.

LanguageEnglish (US)
Pages108-113
Number of pages6
JournalBiochemistry and Biophysics Reports
Volume17
DOIs
StatePublished - Mar 1 2019

Fingerprint

Glucokinase
Hypoglycemia
Necrosis
Adenosine Triphosphate
Cell death
Cell Death
Glucose
Phosphorylation
Cells
Permeability
Cell Survival
Polyomavirus Transforming Antigens
Caspase 7
Mutation
Annexin A5
Viral Tumor Antigens
Caspase 3
Chemical activation
Staining and Labeling

Keywords

  • Beta-cell death
  • Glucokinase
  • Hyperinsulinemic hypoglycemia
  • Hyperinsulinemic hypoglycemia subtype glucokinase
  • Necrosis

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Hyperinsulinemic hypoglycemia subtype glucokinase V91L mutant induces necrosis in β-cells via ATP depletion. / Lu, Brian; Tonne, Jason M.; Munoz-Gomez, Miguel; Ikeda, Yasuhiro H.

In: Biochemistry and Biophysics Reports, Vol. 17, 01.03.2019, p. 108-113.

Research output: Contribution to journalArticle

@article{0865c75ba7cc474ebe0b0ec75055ea5e,
title = "Hyperinsulinemic hypoglycemia subtype glucokinase V91L mutant induces necrosis in β-cells via ATP depletion",
abstract = "Hyperinsulinemic hypoglycemia subtype glucokinase (GCK-HH) is caused by an activating mutation in glucokinase (GCK) and has been shown to increase β-cell death. However, the mechanism of β-cell death in GCK-HH remains poorly understood. Here, we expressed the GCK-HH V91L GCK mutant in INS-1 832/13 cells to determine the effect of the mutation on β-cell viability and the mechanisms of β-cell death. We showed that expression of the V91L GCK mutant in INS-1 832/13 cells resulted in a rapid glucose concentration-dependent loss of cell viability. At 11 mM D-glucose, INS-1 832/13 cells expressing V91L GCK showed increased cell permeability without significant increases in Annexin V staining or caspase 3/7 activation, indicating that these cells are primarily undergoing cell death via necrosis. Over-expression of SV40 large T antigen, which inhibits the p53 pathway, did not affect the V91L GCK-induced cell death. We also found that non-phosphorylatable L-glucose did not induce rapid cell death. Of note, glucose phosphorylation coincided with a 90{\%} loss of intracellular ATP content. Thus, our data suggest that the GCK V91L mutant induces rapid necrosis in INS-1 cells through accelerated glucose phosphorylation, ATP depletion, and increased cell permeability.",
keywords = "Beta-cell death, Glucokinase, Hyperinsulinemic hypoglycemia, Hyperinsulinemic hypoglycemia subtype glucokinase, Necrosis",
author = "Brian Lu and Tonne, {Jason M.} and Miguel Munoz-Gomez and Ikeda, {Yasuhiro H}",
year = "2019",
month = "3",
day = "1",
doi = "10.1016/j.bbrep.2018.12.002",
language = "English (US)",
volume = "17",
pages = "108--113",
journal = "Biochemistry and Biophysics Reports",
issn = "2405-5808",
publisher = "Elsevier BV",

}

TY - JOUR

T1 - Hyperinsulinemic hypoglycemia subtype glucokinase V91L mutant induces necrosis in β-cells via ATP depletion

AU - Lu, Brian

AU - Tonne, Jason M.

AU - Munoz-Gomez, Miguel

AU - Ikeda, Yasuhiro H

PY - 2019/3/1

Y1 - 2019/3/1

N2 - Hyperinsulinemic hypoglycemia subtype glucokinase (GCK-HH) is caused by an activating mutation in glucokinase (GCK) and has been shown to increase β-cell death. However, the mechanism of β-cell death in GCK-HH remains poorly understood. Here, we expressed the GCK-HH V91L GCK mutant in INS-1 832/13 cells to determine the effect of the mutation on β-cell viability and the mechanisms of β-cell death. We showed that expression of the V91L GCK mutant in INS-1 832/13 cells resulted in a rapid glucose concentration-dependent loss of cell viability. At 11 mM D-glucose, INS-1 832/13 cells expressing V91L GCK showed increased cell permeability without significant increases in Annexin V staining or caspase 3/7 activation, indicating that these cells are primarily undergoing cell death via necrosis. Over-expression of SV40 large T antigen, which inhibits the p53 pathway, did not affect the V91L GCK-induced cell death. We also found that non-phosphorylatable L-glucose did not induce rapid cell death. Of note, glucose phosphorylation coincided with a 90% loss of intracellular ATP content. Thus, our data suggest that the GCK V91L mutant induces rapid necrosis in INS-1 cells through accelerated glucose phosphorylation, ATP depletion, and increased cell permeability.

AB - Hyperinsulinemic hypoglycemia subtype glucokinase (GCK-HH) is caused by an activating mutation in glucokinase (GCK) and has been shown to increase β-cell death. However, the mechanism of β-cell death in GCK-HH remains poorly understood. Here, we expressed the GCK-HH V91L GCK mutant in INS-1 832/13 cells to determine the effect of the mutation on β-cell viability and the mechanisms of β-cell death. We showed that expression of the V91L GCK mutant in INS-1 832/13 cells resulted in a rapid glucose concentration-dependent loss of cell viability. At 11 mM D-glucose, INS-1 832/13 cells expressing V91L GCK showed increased cell permeability without significant increases in Annexin V staining or caspase 3/7 activation, indicating that these cells are primarily undergoing cell death via necrosis. Over-expression of SV40 large T antigen, which inhibits the p53 pathway, did not affect the V91L GCK-induced cell death. We also found that non-phosphorylatable L-glucose did not induce rapid cell death. Of note, glucose phosphorylation coincided with a 90% loss of intracellular ATP content. Thus, our data suggest that the GCK V91L mutant induces rapid necrosis in INS-1 cells through accelerated glucose phosphorylation, ATP depletion, and increased cell permeability.

KW - Beta-cell death

KW - Glucokinase

KW - Hyperinsulinemic hypoglycemia

KW - Hyperinsulinemic hypoglycemia subtype glucokinase

KW - Necrosis

UR - http://www.scopus.com/inward/record.url?scp=85058647578&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85058647578&partnerID=8YFLogxK

U2 - 10.1016/j.bbrep.2018.12.002

DO - 10.1016/j.bbrep.2018.12.002

M3 - Article

VL - 17

SP - 108

EP - 113

JO - Biochemistry and Biophysics Reports

T2 - Biochemistry and Biophysics Reports

JF - Biochemistry and Biophysics Reports

SN - 2405-5808

ER -