The regulatory role of hepatoma-derived growth factor as an angiogenic factor in the eye.
OBJECTIVE
Hepatoma-derived growth factor (HDGF) is a mitogen that promotes endothelial proliferation and neuronal survival. Using a unique technology of ligandomics, we recently identified HDGF as a retinal endothelial binding protein. The purpose of this study is to examine the role of HDGF in regulating ocular vasculature and the expression of HDGF in the retina.


METHODS
HDGF expression in the retinal was analyzed with western blot and immunohistochemistry. Angiogenic activity was investigated in human retinal microvascular endothelial cells (HRMVECs) with in vitro endothelial proliferation, migration, and permeability assays. In vivo angiogenic activity was quantified with a corneal pocket assay. The Evans blue assay and western blot using anti-mouse albumin were performed to detect the capacity of HDGF to induce retinal vascular leakage.


RESULTS
Immunohistochemistry revealed that HDGF is expressed in the retina with a distinct pattern. HDGF was detected in retinal ganglion cells and the inner nuclear layer but not in the inner plexiform layer, suggesting that HDGF is expressed in the nucleus, but not in the cytoplasm, of retinal neurons. In contrast to family member HDGF-related protein 3 (HRP-3) that has no expression in photoreceptors, HDGF is also present in the outer nuclear layer and the inner and outer segments of photoreceptors. This suggests that HDGF is expressed in the nucleus as well as the cytoplasm of photoreceptors. In vitro functional assays showed that HDGF induced the proliferation, migration, and permeability of HRMVECs. Corneal pocket assay indicated that HDGF directly stimulated angiogenesis in vivo. Intravitreal injection of HDGF significantly induced retinal vascular leakage.


CONCLUSIONS
These results suggest that HDGF is an angiogenic factor that regulates retinal vasculature in physiologic and pathological conditions. Identification of HDGF by ligandomics and its independent characterization in this study also support the validity of this new technology for systematic identification of cellular ligands, including angiogenic factors.
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL.
Michelle E
ME
LeBlanc
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL.
Weiwen
W
Wang
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL; Department of Ophthalmology, Zhongshan Hospital of Fudan University, Shanghai, China.
Xiuping
X
Chen
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL; Department of Ophthalmology, 2 People's Hospital of Zhengzhou, Zhengzhou, Henan, China.
Yanli
Y
Ji
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL.
Akhalesh
A
Shakya
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL.
Chen
C
Shen
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL; Department of Ophthalmology, Jinan 2 People's Hospital, Jinan, China.
Chenming
C
Zhang
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL.
Vivianne
V
Gonzalez
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL.
Megan
M
Brewer
Department of Physiology, the University of Oklahoma Health Sciences Center, Oklahoma City, OK.
Jian-Xing
JX
Ma
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL.
Rong
R
Wen
Department of Molecular and Cellular Pharmacology, Sylvester Comprehensive Cancer Center, University of Miami, School of Medicine.
Fangliang
F
Zhang
Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL; Vascular Biology Institute, University of Miami School of Medicine, Miami, FL.
Wei
W
Li
D043925:Angiogenesis Inducing Agents / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D002199:Capillary Permeability; D002465:Cell Movement; D049109:Cell Proliferation; D020256:Choroidal Neovascularization / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism*; D005260:Female; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010100:Oxygen / Q000633:toxicity; D060888:Real-Time Polymerase Chain Reaction; D012160:Retina / Q000378:metabolism*; D012171:Retinal Vessels / Q000166:cytology; D012178:Retinopathy of Prematurity / Q000139:chemically induced / Q000378:metabolism*; D014822:Vitreous Body / Q000378:metabolism
D043925:Angiogenesis Inducing Agents; D036341:Intercellular Signaling Peptides and Proteins; C051610:hepatoma-derived growth factor; D010100:Oxygen