Notch; VEGF; Skeletal muscle; Therapeutic angiogenesis; Ephrins; Gene therapy; Intussusception
Gianni-Barrera R., Butschkau A., Uccelli A., Certelli A., Valente P., Bartolomeo M., Groppa E., Burger M. G., Hlushchuk R., Heberer M., Schaefer D. J., Gürke L., Djonov V., Vollmar B., Banfi A. (2018), PDGF-BB regulates splitting angiogenesis in skeletal muscle by limiting VEGF-induced endothelial proliferation, in Angiogenesis
, 21(4), 883-900.
Melly Ludovic, Cerino Giulia, Frobert Aurélien, Cook Stéphane, Giraud Marie-Noëlle, Carrel Thierry, Tevaearai Stahel Hendrik T., Eckstein Friedrich, Rondelet Benoît, Marsano Anna, Banfi Andrea (2018), Myocardial infarction stabilization by cell-based expression of controlled Vascular Endothelial Growth Factor levels, in Journal of Cellular and Molecular Medicine
, 22(5), 2580-2591.
Groppa Elena, Brkic Sime, Uccelli Andrea, Wirth Galina, Korpisalo‐Pirinen Petra, Filippova Maria, Dasen Boris, Sacchi Veronica, Muraro Manuele Giuseppe, Trani Marianna, Reginato Silvia, Gianni‐Barrera Roberto, Ylä‐Herttuala Seppo, Banfi Andrea (2018), EphrinB2/EphB4 signaling regulates non‐sprouting angiogenesis by VEGF, in EMBO reports
, 19(5), e45054-e45054.
Grosso Andrea, Burger Maximilian G., Lunger Alexander, Schaefer Dirk J., Banfi Andrea, Di Maggio Nunzia (2017), It Takes Two to Tango: Coupling of Angiogenesis and Osteogenesis for Bone Regeneration, in Frontiers in Bioengineering and Biotechnology
, 5, 68.
Boccardo Stefano, Gaudiello Emanuele, Melly Ludovic, Cerino Giulia, Ricci Davide, Martin Ivan, Eckstein Friedrich, Banfi Andrea, Marsano Anna (2016), Engineered mesenchymal cell-based patches as controlled VEGF delivery systems to induce extrinsic angiogenesis, in Acta Biomaterialia
, 42, 127-135.
Gianni-Barrera Roberto, Burger Maximilian, Wolff Thomas, Heberer Michael, Schaefer Dirk J., Gürke Lorenz, Mujagic Edin, Banfi Andrea (2016), Long-term safety and stability of angiogenesis induced by balanced single-vector co-expression of PDGF-BB and VEGF164 in skeletal muscle, in Scientific Reports
, 6(1), 21546-21546.
Background. Ischemic cardiovascular disease is the most common cause of death in the Western world and, despite advances in medical and surgical therapy, the morbidity and mortality remain very high. Therapeutic angiogenesis aims to induce the formation of new blood vessels to improve the perfusion of ischemic tissue in patients with end-stage coronary artery or peripheral arterial disease that are not amenable to other treatment options. Vascular Endothelial Growth Factor-A (VEGF) is the most potent and specific angiogenic factor and it has been tested in several clinical trials with a variety of delivery methods. However, preclinical studies clearly demonstrated that high doses of VEGF can also induce aberrant vascular structures which resemble cavernous hemangiomas and grow progressively. The results of placebo-controlled clinical trials have been disappointing and yielded mostly negative results. The main conundrum of VEGF delivery-based strategies for therapeutic angiogenesis is its apparently limited therapeutic window in vivo, such that low doses are safe, but mostly inefficient, and higher doses become rapidly unsafe.Rationale. Our previous results show that the transition between normal and aberrant angiogenesis takes place in an all-or-none fashion across a discrete threshold level of VEGF expression. However, we found that this threshold is not an intrinsic property of VEGF dose alone, but rather depends on the balance between angiogenic stimulation by VEGF and vascular maturation by PDGF-BB-mediated pericyte recruitment. Our results in the previous funding period show that 1) VEGF overexpression in skeletal muscle at therapeutic doses induces angiogenesis by circumferential enlargement followed by intussusception and not by sprouting; 2) Notch-1 regulates the initial stages of vascular enlargement by a different pattern of activation compared to sprouting, but is not involved in intussusceptive remodeling; 3) EphrinB2/EphB4 signaling controls the switch between normal and aberrant angiogenesis by different VEGF doses, by regulating the degree of endothelial proliferation and initial vascular enlargement.Specific aims. In the current funding period we propose to extend these results to investigate: 1) the role of Notch4 signaling in the switch between normal and aberrant angiogenesis by VEGF in skeletal muscle; 2) the cross-talk between EphrinB2/EphB4 signaling and Notch4 activation; and 3) whether stimulation of EphB4 signaling can improve the safety and therapeutic efficacy of VEGF gene delivery in a murine hind-limb ischemia model.Experimental design. Monoclonal populations of retrovirally transduced myoblast, which stably secrete different amounts of VEGF, or a highly controlled fibrin-based platform for protein delivery, will be used to deliver specific VEGF doses in skeletal muscle. Notch4-/- mice will be used to study loss of Notch4 function, while systemic treatment with specific molecules will be used to stimulate the Notch pathway, or block or activate EphB4 signaling.Expected value of the proposed project. The experiments proposed are expected to: 1) provide fundamental knowledge on the basic mechanisms by which VEGF dose determines a switch between normal and aberrant angiogenesis in skeletal muscle, and in particular the regulation of VEGF-induced intussusceptive angiogenesis, which is a poorly understood, but therapeutically relevant process; and 2) test the potential of targeting the EphrinB2/EphB4 pathway to improve both the efficacy and safety of therapeutic angiogenesis by VEGF delivery in a functional ischemia model and with a clinically relevant gene therapy approach.