Endothelial PFKFB3 plays a critical role in angiogenesis.
OBJECTIVE
Vascular cells, particularly endothelial cells, adopt aerobic glycolysis to generate energy to support cellular functions. The effect of endothelial glycolysis on angiogenesis remains unclear. 6-Phosphofructo-2-kinase/fructose-2, 6-bisphosphatase, isoform 3 (PFKFB3) is a critical enzyme for endothelial glycolysis. By blocking or deleting PFKFB3 in endothelial cells, we investigated the influence of endothelial glycolysis on angiogenesis both in vitro and in vivo.


RESULTS
Under hypoxic conditions or after treatment with angiogenic factors, endothelial PFKFB3 was upregulated both in vitro and in vivo. The knockdown or overexpression of PFKFB3 suppressed or accelerated endothelial proliferation and migration in vitro, respectively. Neonatal mice from a model of oxygen-induced retinopathy showed suppressed neovascular growth in the retina when endothelial PFKFB3 was genetically deleted or when the mice were treated with a PFKFB3 inhibitor. In addition, tumors implanted in mice deficient in endothelial PFKFB3 grew more slowly and were provided with less blood flow. A lower level of phosphorylated protein kinase B was observed in PFKFB3-knockdown endothelial cells, which was accompanied by a decrease in intracellular lactate. The addition of lactate to PFKFB3-knockdown cells rescued the suppression of endothelial proliferation and migration.


CONCLUSIONS
The blockade or deletion of endothelial PFKFB3 decreases angiogenesis both in vitro and in vivo. Thus, PFKFB3 is a promising target for the reduction of endothelial glycolysis and its related pathological angiogenesis.
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Yiming
Y
Xu
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Xiaofei
X
An
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Xin
X
Guo
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Tsadik Ghebreamlak
TG
Habtetsion
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Yong
Y
Wang
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Xizhen
X
Xu
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Sridhar
S
Kandala
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Qinkai
Q
Li
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Honggui
H
Li
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Chunxiang
C
Zhang
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Ruth B
RB
Caldwell
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
David J
DJ
Fulton
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Yunchao
Y
Su
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Md Nasrul
MN
Hoda
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.).
Gang
G
Zhou
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.). yhuo@gru.edu cdwu@tamu.edu.
Chaodong
C
Wu
From the Vascular Biology Center, Department of Cellular Biology and Anatomy (Y.X., X.A., X.X., S.K., R.B.C., D.J.F., Y.H.), Cancer Center, Department of Medicine (T.G.H., G.Z.), and Department of Pharmacology and Toxicology (Y.W., Y.S.), Medical College of Georgia and Departments of Medical Laboratory, Imaging and Radiologic Sciences, and Neurology (M.N.H.), Georgia Regents University, Augusta; Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China (X.A., Q.L., Y.H.); Department of Nutrition and Food Science, Texas A&M University, College Station (X.G., H.L., C.W.); and Department of Pharmacology, Rush Medical College, Rush University, Chicago, IL (C.Z.). yhuo@gru.edu cdwu@tamu.edu.
Yuqing
Y
Huo
D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000502:physiology*; D005260:Female; D006019:Glycolysis; D006801:Humans; D019344:Lactic Acid / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009389:Neovascularization, Pathologic / Q000209:etiology*; D025481:Phosphofructokinase-2 / Q000502:physiology*; D051057:Proto-Oncogene Proteins c-akt / Q000502:physiology; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology
D042461:Vascular Endothelial Growth Factor A; D019344:Lactic Acid; C107345:PFKFB3 protein, human; D025481:Phosphofructokinase-2; D051057:Proto-Oncogene Proteins c-akt
angiogenesis; anoxia; endothelial cells; glycolysis