VEGF dose regulates vascular stabilization through Semaphorin3A and the Neuropilin-1+ monocyte/TGF-β1 paracrine axis.
VEGF is widely investigated for therapeutic angiogenesis, but while short-term delivery is desirable for safety, it is insufficient for new vessel persistence, jeopardizing efficacy. Here, we investigated whether and how VEGF dose regulates nascent vessel stabilization, to identify novel therapeutic targets. Monoclonal populations of transduced myoblasts were used to homogeneously express specific VEGF doses in SCID mouse muscles. VEGF was abrogated after 10 and 17 days by Aflibercept treatment. Vascular stabilization was fastest with low VEGF, but delayed or prevented by higher doses, without affecting pericyte coverage. Rather, VEGF dose-dependently inhibited endothelial Semaphorin3A expression, thereby impairing recruitment of Neuropilin-1-expressing monocytes (NEM), TGF-β1 production and endothelial SMAD2/3 activation. TGF-β1 further initiated a feedback loop stimulating endothelial Semaphorin3A expression, thereby amplifying the stabilizing signals. Blocking experiments showed that NEM recruitment required endogenous Semaphorin3A and that TGF-β1 was necessary to start the Semaphorin3A/NEM axis. Conversely, Semaphorin3A treatment promoted NEM recruitment and vessel stabilization despite high VEGF doses or transient adenoviral delivery. Therefore, VEGF inhibits the endothelial Semaphorin3A/NEM/TGF-β1 paracrine axis and Semaphorin3A treatment accelerates stabilization of VEGF-induced angiogenesis.
Department of Biomedicine, University of Basel, Basel, Switzerland Department of Surgery, Basel University Hospital, Basel, Switzerland.
Elena
E
Groppa
Department of Biomedicine, University of Basel, Basel, Switzerland Department of Surgery, Basel University Hospital, Basel, Switzerland.
Sime
S
Brkic
Department of Biomedicine, University of Basel, Basel, Switzerland Department of Surgery, Basel University Hospital, Basel, Switzerland.
Emmanuela
E
Bovo
Department of Biomedicine, University of Basel, Basel, Switzerland Department of Surgery, Basel University Hospital, Basel, Switzerland.
Silvia
S
Reginato
Department of Biomedicine, University of Basel, Basel, Switzerland Department of Surgery, Basel University Hospital, Basel, Switzerland.
Veronica
V
Sacchi
Department of Biomedicine, University of Basel, Basel, Switzerland Department of Surgery, Basel University Hospital, Basel, Switzerland.
Nunzia
N
Di Maggio
Department of Biomedicine, University of Basel, Basel, Switzerland Department of Surgery, Basel University Hospital, Basel, Switzerland.
Manuele G
MG
Muraro
Department of Biomedicine, University of Basel, Basel, Switzerland.
Diego
D
Calabrese
Department of Biomedicine, University of Basel, Basel, Switzerland Department of Surgery, Basel University Hospital, Basel, Switzerland.
Michael
M
Heberer
Department of Biomedicine, University of Basel, Basel, Switzerland Department of Surgery, Basel University Hospital, Basel, Switzerland.
Roberto
R
Gianni-Barrera
Department of Biomedicine, University of Basel, Basel, Switzerland Department of Surgery, Basel University Hospital, Basel, Switzerland andrea.banfi@usb.ch.
Andrea
A
Banfi
D043925:Angiogenesis Inducing Agents / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D020104:Immunophilins / Q000378:metabolism*; D051379:Mice; D016513:Mice, SCID; D032446:Myoblasts / Q000378:metabolism / Q000502:physiology; D019899:Paracrine Communication; D040121:Semaphorin-3A / Q000378:metabolism*; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism* / Q000494:pharmacology
D043925:Angiogenesis Inducing Agents; D040121:Semaphorin-3A; D051899:Smad2 Protein; D051900:Smad3 Protein; D053773:Transforming Growth Factor beta1; D042461:Vascular Endothelial Growth Factor A; C467480:vascular endothelial growth factor A, mouse; D020104:Immunophilins
VEGF; monocyte; TGF‐β1; semaphorin3A; vascular stabilization