Expression of the oxygen-regulated protein ORP150 accelerates wound healing by modulating intracellular VEGF transport
Kentaro Ozawa, … , Satoshi Ogawa, Tohru Ohshima
Kentaro Ozawa, … , Satoshi Ogawa, Tohru Ohshima
Published July 1, 2001
Citation Information: J Clin Invest. 2001;108(1):41-50. https://doi.org/10.1172/JCI11772.
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Article

Expression of the oxygen-regulated protein ORP150 accelerates wound healing by modulating intracellular VEGF transport

Abstract

Expression of angiogenic factors such as VEGF under conditions of hypoxia or other kinds of cell stress contributes to neovascularization during wound healing. The inducible endoplasmic reticulum chaperone oxygen-regulated protein 150 (ORP150) is expressed in human wounds along with VEGF. Colocalization of these two molecules was observed in macrophages in the neovasculature, suggesting a role of ORP150 in the promotion of angiogenesis. Local administration of ORP150 sense adenovirus to wounds of diabetic mice, a treatment that efficiently targeted this gene product to the macrophages of wound beds, increased VEGF antigen in wounds and accelerated repair and neovascularization. In cultured human macrophages, inhibition of ORP150 expression caused retention of VEGF antigen within the endoplasmic reticulum (ER), while overexpression of ORP150 promoted the secretion of VEGF into hypoxic culture supernatants. Taken together, these data suggest an important role for ORP150 in the setting of impaired wound repair and identify a key, inducible chaperone-like molecule in the ER. This novel facet of the angiogenic response may be amenable to therapeutic manipulation.

Authors

Kentaro Ozawa, Toshikazu Kondo, Osamu Hori, Yasuko Kitao, David M. Stern, Wolfgang Eisenmenger, Satoshi Ogawa, Tohru Ohshima

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Characterization of wounds infected with adenovirus. (a–h) Two days afte...
Characterization of wounds infected with adenovirus. (a–h) Two days after the introduction of a wound (6 mm in diameter) in db/db mice, a mixture of adenovirus (AxGFP and Ad/S-ORP150; 5 × 107 pfu each) was administered at each wound, as described. Two days after the infection, the tissue was sampled and subjected to H&E staining (×4) (a), immunostaining with anti-F4/80 Ab (×40) (b), or visualization of GFP signal by fluorescent microscopy (×40) (c). Signals derived from F4/80 and GFP were digitally overlapped (×40) (d). (a) Filled arrowheads indicate the initial wound area, and the open arrowheads indicate the edge of the granulation tissue. (e–h) Granulation tissue was stained with anti-ORP150 Ab and signals derived from (e) GFP (×40) and (f) ORP150 (×40) were visualized by fluorescent microscopy. (g) Signals of GFP and ORP150 were digitally overlapped. (×40.) (h) The adjacent section was immunostained with anti-F4/80 Ab. (×40.) (i–k) Either the indicated amount of AxGFP and Ad/S-ORP150 (open bars) or (i and j) AxCALacZ and Ad/S-ORP150 (108 pfu each; filled bars), or (k) AxCALacZ and AxGFP (108 pfu each; filled bar) was administered, as described. Three days after the infection, either GFP-positive cells (i) or the percentage of GFP-positive cells in F4/80-positive cells (j) was counted in the granulation tissue as described in the text. The content of tissue ORP150 antigen was assessed by ELISA as described. (k). For i–k, n = 6, mean ± SD. *P < 0.01 by multiple comparison analysis.