Ablation of 4E-BP1/2 prevents hyperglycemia-mediated induction of VEGF expression in the rodent retina and in Müller cells in culture

Tabitha L. Schrufer, David A. Antonetti, Nahum Sonenberg, Scot R. Kimball, Thomas W. Gardner, Leonard S. Jefferson

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

OBJECTIVE - Vascular endothelial growth factor (VEGF) contributes to diabetic retinopathy, but control of its expression is not well understood. Here, we tested the hypothesis that hyperglycemia mediates induction of VEGF expression in a eukaryotic initiation factor 4E (eIF4E) binding protein (4E-BP) 1 and 2 dependent manner. RESEARCH DESIGN AND METHODS - The retina was harvested from control and type 1 diabetic rats and mice and analyzed for VEGF mRNA and protein expression as well as biomarkers of translational control mechanisms. Similar analyses were performed in Müller cell cultures exposed to hyperglycemic conditions. The effect of 4E-BP1 and 4E-BP2 gene deletion on VEGF expression was examined in mice and in mouse embryo fibroblasts (MEFs). RESULTS - Whereas VEGF mRNA in the retina remained constant, VEGF expression was increased as early as 2 weeks after the onset of diabetes. Increases in expression of 4E-BP1 protein mirrored those of VEGF and expression of 4E-BP1 mRNA was unchanged. Similar results were observed after 10 h of exposure of cells in culture to hyperglycemic conditions. Importantly, the diabetes-induced increase in VEGF expression was not observed in mice deficient in 4E-BP1 and 4E-BP2, nor in MEFs lacking the two proteins. CONCLUSIONS - Hyperglycemia induces VEGF expression through cap-independent mRNA translation mediated by increased expression of 4E-BP1. Because the VEGF mRNA contains two internal ribosome entry sites, the increased expression is likely a consequence of ribosome loading at these sites. These findings provide new insights into potential targets for treatment of diabetic retinopathy.

Original languageEnglish (US)
Pages (from-to)2107-2116
Number of pages10
JournalDiabetes
Volume59
Issue number9
DOIs
StatePublished - Sep 2010

Fingerprint

Hyperglycemia
Vascular Endothelial Growth Factor A
Retina
Rodentia
Cell Culture Techniques
Messenger RNA
Diabetic Retinopathy
Embryonic Structures
Fibroblasts
Eukaryotic Initiation Factor-4E
Proteins
Gene Deletion
Protein Biosynthesis
Ribosomes
Carrier Proteins
Research Design
Biomarkers

All Science Journal Classification (ASJC) codes

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

Cite this

Schrufer, Tabitha L. ; Antonetti, David A. ; Sonenberg, Nahum ; Kimball, Scot R. ; Gardner, Thomas W. ; Jefferson, Leonard S. / Ablation of 4E-BP1/2 prevents hyperglycemia-mediated induction of VEGF expression in the rodent retina and in Müller cells in culture. In: Diabetes. 2010 ; Vol. 59, No. 9. pp. 2107-2116.
@article{77d1b670ba1042b3aa40086a02dab248,
title = "Ablation of 4E-BP1/2 prevents hyperglycemia-mediated induction of VEGF expression in the rodent retina and in M{\"u}ller cells in culture",
abstract = "OBJECTIVE - Vascular endothelial growth factor (VEGF) contributes to diabetic retinopathy, but control of its expression is not well understood. Here, we tested the hypothesis that hyperglycemia mediates induction of VEGF expression in a eukaryotic initiation factor 4E (eIF4E) binding protein (4E-BP) 1 and 2 dependent manner. RESEARCH DESIGN AND METHODS - The retina was harvested from control and type 1 diabetic rats and mice and analyzed for VEGF mRNA and protein expression as well as biomarkers of translational control mechanisms. Similar analyses were performed in M{\"u}ller cell cultures exposed to hyperglycemic conditions. The effect of 4E-BP1 and 4E-BP2 gene deletion on VEGF expression was examined in mice and in mouse embryo fibroblasts (MEFs). RESULTS - Whereas VEGF mRNA in the retina remained constant, VEGF expression was increased as early as 2 weeks after the onset of diabetes. Increases in expression of 4E-BP1 protein mirrored those of VEGF and expression of 4E-BP1 mRNA was unchanged. Similar results were observed after 10 h of exposure of cells in culture to hyperglycemic conditions. Importantly, the diabetes-induced increase in VEGF expression was not observed in mice deficient in 4E-BP1 and 4E-BP2, nor in MEFs lacking the two proteins. CONCLUSIONS - Hyperglycemia induces VEGF expression through cap-independent mRNA translation mediated by increased expression of 4E-BP1. Because the VEGF mRNA contains two internal ribosome entry sites, the increased expression is likely a consequence of ribosome loading at these sites. These findings provide new insights into potential targets for treatment of diabetic retinopathy.",
author = "Schrufer, {Tabitha L.} and Antonetti, {David A.} and Nahum Sonenberg and Kimball, {Scot R.} and Gardner, {Thomas W.} and Jefferson, {Leonard S.}",
year = "2010",
month = "9",
doi = "10.2337/db10-0148",
language = "English (US)",
volume = "59",
pages = "2107--2116",
journal = "Diabetes",
issn = "0012-1797",
publisher = "American Diabetes Association Inc.",
number = "9",

}

Ablation of 4E-BP1/2 prevents hyperglycemia-mediated induction of VEGF expression in the rodent retina and in Müller cells in culture. / Schrufer, Tabitha L.; Antonetti, David A.; Sonenberg, Nahum; Kimball, Scot R.; Gardner, Thomas W.; Jefferson, Leonard S.

In: Diabetes, Vol. 59, No. 9, 09.2010, p. 2107-2116.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Ablation of 4E-BP1/2 prevents hyperglycemia-mediated induction of VEGF expression in the rodent retina and in Müller cells in culture

AU - Schrufer, Tabitha L.

AU - Antonetti, David A.

AU - Sonenberg, Nahum

AU - Kimball, Scot R.

AU - Gardner, Thomas W.

AU - Jefferson, Leonard S.

PY - 2010/9

Y1 - 2010/9

N2 - OBJECTIVE - Vascular endothelial growth factor (VEGF) contributes to diabetic retinopathy, but control of its expression is not well understood. Here, we tested the hypothesis that hyperglycemia mediates induction of VEGF expression in a eukaryotic initiation factor 4E (eIF4E) binding protein (4E-BP) 1 and 2 dependent manner. RESEARCH DESIGN AND METHODS - The retina was harvested from control and type 1 diabetic rats and mice and analyzed for VEGF mRNA and protein expression as well as biomarkers of translational control mechanisms. Similar analyses were performed in Müller cell cultures exposed to hyperglycemic conditions. The effect of 4E-BP1 and 4E-BP2 gene deletion on VEGF expression was examined in mice and in mouse embryo fibroblasts (MEFs). RESULTS - Whereas VEGF mRNA in the retina remained constant, VEGF expression was increased as early as 2 weeks after the onset of diabetes. Increases in expression of 4E-BP1 protein mirrored those of VEGF and expression of 4E-BP1 mRNA was unchanged. Similar results were observed after 10 h of exposure of cells in culture to hyperglycemic conditions. Importantly, the diabetes-induced increase in VEGF expression was not observed in mice deficient in 4E-BP1 and 4E-BP2, nor in MEFs lacking the two proteins. CONCLUSIONS - Hyperglycemia induces VEGF expression through cap-independent mRNA translation mediated by increased expression of 4E-BP1. Because the VEGF mRNA contains two internal ribosome entry sites, the increased expression is likely a consequence of ribosome loading at these sites. These findings provide new insights into potential targets for treatment of diabetic retinopathy.

AB - OBJECTIVE - Vascular endothelial growth factor (VEGF) contributes to diabetic retinopathy, but control of its expression is not well understood. Here, we tested the hypothesis that hyperglycemia mediates induction of VEGF expression in a eukaryotic initiation factor 4E (eIF4E) binding protein (4E-BP) 1 and 2 dependent manner. RESEARCH DESIGN AND METHODS - The retina was harvested from control and type 1 diabetic rats and mice and analyzed for VEGF mRNA and protein expression as well as biomarkers of translational control mechanisms. Similar analyses were performed in Müller cell cultures exposed to hyperglycemic conditions. The effect of 4E-BP1 and 4E-BP2 gene deletion on VEGF expression was examined in mice and in mouse embryo fibroblasts (MEFs). RESULTS - Whereas VEGF mRNA in the retina remained constant, VEGF expression was increased as early as 2 weeks after the onset of diabetes. Increases in expression of 4E-BP1 protein mirrored those of VEGF and expression of 4E-BP1 mRNA was unchanged. Similar results were observed after 10 h of exposure of cells in culture to hyperglycemic conditions. Importantly, the diabetes-induced increase in VEGF expression was not observed in mice deficient in 4E-BP1 and 4E-BP2, nor in MEFs lacking the two proteins. CONCLUSIONS - Hyperglycemia induces VEGF expression through cap-independent mRNA translation mediated by increased expression of 4E-BP1. Because the VEGF mRNA contains two internal ribosome entry sites, the increased expression is likely a consequence of ribosome loading at these sites. These findings provide new insights into potential targets for treatment of diabetic retinopathy.

UR - http://www.scopus.com/inward/record.url?scp=77956374138&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77956374138&partnerID=8YFLogxK

U2 - 10.2337/db10-0148

DO - 10.2337/db10-0148

M3 - Article

C2 - 20547975

AN - SCOPUS:77956374138

VL - 59

SP - 2107

EP - 2116

JO - Diabetes

JF - Diabetes

SN - 0012-1797

IS - 9

ER -