Keiichi Park1, Hideki Amano2, Yoshiya Ito3, Shinya Kashiwagi4, Yasuharu Yamazaki4, Akira Takeda4, Masabumi Shibuya5, Hidero Kitasato6, Masataka Majima7. 1. Department of Pharmacology, Kitasato University School of Medicine, Kanagawa, Japan; Department of Plastic and Aesthetic Surgery, Kitasato University School of Medicine, Kanagawa, Japan. 2. Department of Pharmacology, Kitasato University School of Medicine, Kanagawa, Japan. 3. Department of Surgery, Kitasato University School of Medicine, Kanagawa, Japan. 4. Department of Plastic and Aesthetic Surgery, Kitasato University School of Medicine, Kanagawa, Japan. 5. Gakubunkan Institute of Physiology and Medicine, Jobu University, Tokyo, Japan. 6. Department of Microbiology, Kitasato University School of Allied Health Science, Kanagawa, Japan. 7. Department of Pharmacology, Kitasato University School of Medicine, Kanagawa, Japan. Electronic address: mmajima@med.kitasto-u.ac.jp.
Abstract
BACKGROUND: Vascular endothelial growth factor (VEGF)-A binds to both VEGF receptor (VEGFR)-1 and VEGFR-2, thereby promoting angiogenesis. It is widely accepted that VEGF-A, especially VEGFR-2, is a central player in angiogenesis, however the role of VEGFR-1 in angiogenesis remains unclear. The present study was conducted to examine the role of VEGFR-1 signaling in angiogenesis, using a quantitative in vivo angiogenesis model. METHODS: Polyurethane sponge disks were implanted into dorsal subcutaneous tissue of mice. Angiogenesis was estimated by determining the number of CD31(+) vessels by immunohistochemical analysis. The expression of pro-angiogenic factors was quantified by reverse transcription quantitative polymerase chain reaction. RESULTS: Compared to control IgG-treated mice, the number of CD31(+) vessels in the sponge implant was significantly suppressed in anti-VEGF-A neutralizing antibody-treated mice. CD31(+) vessel counts were suppressed in VEGFR-1 tyrosine kinase knockout (TKKO) mice, at the same level as in VEGFR-2 tyrosine kinase inhibitor (ZD6474)-treated mice compared to wild-type (WT) mice. The accumulation of VEGFR-1(+) cells in granulation tissue was significantly suppressed in VEGFR-1 TKKO mice compared to WT mice. In addition, expression of the pro-angiogenic growth factors, VEGF-A, matrix metalloproteinase-2, interleukin-6, and basic fibroblast growth factor in granulation tissue was suppressed in VEGFR-1 TKKO mice. A bone marrow (BM) transplantation experiment showed that the number of VEGFR-1(+) BM-derived cells and angiogenesis were significantly suppressed in VEGFR-1 TKKO mice transplanted with green fluorescent protein (GFP)(+) VEGFR-1 TKKO BM compared to WT mice transplanted with GFP(+) WT BM. CONCLUSIONS: These results suggest that the VEGFR-1 tyrosine kinase signaling has an effect on angiogenesis. A selective VEGFR-1 agonist/antagonist could be a candidate therapeutic agent to control angiogenesis with recruitment of BM cells.
BACKGROUND:Vascular endothelial growth factor (VEGF)-A binds to both VEGF receptor (VEGFR)-1 and VEGFR-2, thereby promoting angiogenesis. It is widely accepted that VEGF-A, especially VEGFR-2, is a central player in angiogenesis, however the role of VEGFR-1 in angiogenesis remains unclear. The present study was conducted to examine the role of VEGFR-1 signaling in angiogenesis, using a quantitative in vivo angiogenesis model. METHODS:Polyurethane sponge disks were implanted into dorsal subcutaneous tissue of mice. Angiogenesis was estimated by determining the number of CD31(+) vessels by immunohistochemical analysis. The expression of pro-angiogenic factors was quantified by reverse transcription quantitative polymerase chain reaction. RESULTS: Compared to control IgG-treated mice, the number of CD31(+) vessels in the sponge implant was significantly suppressed in anti-VEGF-A neutralizing antibody-treated mice. CD31(+) vessel counts were suppressed in VEGFR-1 tyrosine kinase knockout (TKKO) mice, at the same level as in VEGFR-2 tyrosine kinase inhibitor (ZD6474)-treated mice compared to wild-type (WT) mice. The accumulation of VEGFR-1(+) cells in granulation tissue was significantly suppressed in VEGFR-1 TKKO mice compared to WT mice. In addition, expression of the pro-angiogenic growth factors, VEGF-A, matrix metalloproteinase-2, interleukin-6, and basic fibroblast growth factor in granulation tissue was suppressed in VEGFR-1 TKKO mice. A bone marrow (BM) transplantation experiment showed that the number of VEGFR-1(+) BM-derived cells and angiogenesis were significantly suppressed in VEGFR-1 TKKO mice transplanted with green fluorescent protein (GFP)(+) VEGFR-1 TKKO BM compared to WT mice transplanted with GFP(+) WT BM. CONCLUSIONS: These results suggest that the VEGFR-1 tyrosine kinase signaling has an effect on angiogenesis. A selective VEGFR-1 agonist/antagonist could be a candidate therapeutic agent to control angiogenesis with recruitment of BM cells.
Authors: Shuang Li; Christian G M van Dijk; Jan Meeldijk; Helena M Kok; Isabelle Blommestein; Annick L F Verbakel; Marit Kotte; Roel Broekhuizen; Miangela M Laclé; Roel Goldschmeding; Caroline Cheng; Niels Bovenschen Journal: Front Oncol Date: 2021-06-09 Impact factor: 6.244