Therapeutic angiogenesis; Gene therapy; Pericytes; VEGF; PDGF-BB; Notch; Ephrins; Angiopoietins; TGF; Angiogenesis;; Pericytes;; Gene therapy;; TGFb.
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, 21546-21546.
Groppa E., Brkic S., Bovo E., Reginato S., Sacchi V., Di Maggio N., Muraro M. G., Calabrese D., Heberer M., Gianni-Barrera R., Banfi A. (2015), VEGF dose regulates vascular stabilization through Semaphorin3A and the Neuropilin-1+ monocyte/TGF- 1 paracrine axis, in EMBO Molecular Medicine
, 7(10), 1366-1384.
Sacchi V., Mittermayr R., Hartinger J., Martino M. M., Lorentz K. M., Wolbank S., Hofmann A., Largo R. A., Marschall J. S., Groppa E., Gianni-Barrera R., Ehrbar M., Hubbell J. A., Redl H., Banfi A. (2014), Long-lasting fibrin matrices ensure stable and functional angiogenesis by highly tunable, sustained delivery of recombinant VEGF164, in Proceedings of the National Academy of Sciences
, 111(19), 6952-6957.
Mujagic Edin, Gianni-Barrera Roberto, Trani Marianna, Patel Abdulsamie, Gürke Lorenz, Heberer Michael, Wolff Thomas, Banfi Andrea (2013), Induction of aberrant vascular growth, but not of normal angiogenesis, by cell-based expression of different doses of human and mouse VEGF is species-dependent., in Human gene therapy methods
, 24(1), 28-37.
Helmrich Uta, Di Maggio Nunzia, Güven Sinan, Groppa Elena, Melly Ludovic, Largo Rene D., Heberer Michael, Martin Ivan, Scherberich Arnaud, Banfi Andrea (2013), Osteogenic graft vascularization and bone resorption by VEGF-expressing human mesenchymal progenitors, in Biomaterials
, 34(21), 5025-5035.
Marsano Anna, Maidhof Robert, Luo Jianwen, Fujikara Kana, Konofagou Elisa, Banfi Andrea, Vunjak-Novakovic Gordana (2013), The effect of controlled expression of VEGF by transduced myoblasts in a cardiac patch on vascularization in a mouse model of myocardial infarction, in Biomaterials
, 34(2), 393-401.
Gianni-Barrera Roberto, Trani Marianna, Fontanellaz Christian, Heberer Michael, Djonov Valentin, Hlushchuk Ruslan, Banfi Andrea (2013), VEGF over-expression in skeletal muscle induces angiogenesis by intussusception rather than sprouting., in Angiogenesis
, 16(1), 123-36.
Melly Ludovic, Boccardo Stefano, Eckstein Friedrich, Banfi Andrea, Marsano Anna (2012), Cell and Gene Therapy Approaches for Cardiac Vascularization, in Cells
, 1, 961-975.
Melly Ludovic F, Marsano Anna, Frobert Aurelien, Boccardo Stefano, Helmrich Uta, Heberer Michael, Eckstein Friedrich S, Carrel Thierry P, Giraud Marie-Noëlle, Tevaearai Hendrik T, Banfi Andrea (2012), Controlled angiogenesis in the heart by cell-based expression of specific vascular endothelial growth factor levels., in Human gene therapy methods
, 23(5), 346-356.
Wolff Thomas, Mujagic Edin, Gianni-Barrera Roberto, Fueglistaler Philipp, Helmrich Uta, Misteli Heidi, Gurke Lorenz, Heberer Michael, Banfi Andrea (2012), FACS-purified myoblasts producing controlled VEGF levels induce safe and stable angiogenesis in chronic hind limb ischemia., in Journal of cellular and molecular medicine
, 16(1), 107-117.
Helmrich Uta, Marsano Anna, Melly Ludovic, Wolff Thomas, Christ Liliane, Heberer Michael, Scherberich Arnaud, Martin Ivan, Banfi Andrea (2012), Generation of human MSC expressing defined xenogenic VEGF levels by optimized transduction and flow cytometry purification., in Tissue engineering. Part C, Methods
, 18(4), 283-292.
Banfi Andrea, von Degenfeld Georges, Gianni-Barrera Roberto, Reginato Silvia, Merchant Milton J, McDonald Donald M, Blau Helen M (2012), Therapeutic angiogenesis due to balanced single-vector delivery of VEGF and PDGF-BB., in FASEB journal : official publication of the Federation of American Societies for Experimental Biolog
, 26, 2486-2497.
Reginato Silvia, Gianni-Barrera Roberto, Banfi Andrea (2011), Taming of the wild vessel: promoting vessel stabilization for safe therapeutic angiogenesis., in Biochemical Society transactions
, 39(6), 1654-1658.
Gianni-Barrera Roberto, Trani Marianna, Reginato Silvia, Banfi Andrea (2011), To sprout or to split? VEGF, Notch and vascular morphogenesis., in Biochemical Society transactions
, 39(6), 1644-1648.
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 coordinated co-expression of PDGF-BB and pericyte recruitment effectively improves both safety and efficacy of VEGF gene delivery, by inducing robust normal and homogeneous angiogenesis despite heterogeneous and high VEGF levels.Specific aims. In the current funding period we propose to extend these results along two lines: 1) to apply these biological concepts in a clinically applicable gene therapy strategy for therapeutic angiogenesis, and 2) to investigate the mechanism underlying the modulation of VEGF effects by PDGF-BB co-expressio. We will dissect the role of pericytes and specific pericyte-mediated signaling pathways in the normalization of VEGF-induced angiogenesis by PDGF-BB (Aim 1). Further, we will investigate whether dose-dependent co-expression prevents angioma growth and leads instead to normal capillary networks by regulating the signaling pathways that control the endothelial cell fate decision to become a sprouting tip cell or a circumferentially growing stalk cell (Aim 2). We will then test the feasibility of AV and AAV-based coordinated co-expression of VEGF and PDGF-BB as gene therapy strategies to achieve safe and efficacious angiogenesis (Aim 3).Experimental design. Monoclonal populations of retrovirally-transduced myoblast, which stably secrete different amounts of VEGF or VEGF+PDGF, will be used to achieve specific expression levels in skeletal muscle. We will then co-express specific molecules to block or activate specific pathways involved in the pericyte-endothelium and endothelium-endothelium cross talk. Finally we will test the applicability of VEGF+PDGF co-delivery to a clinically relevant delivery system using a bicistronic AV or AAV vector.Expected value of the proposed project. The proposed research is expected to provide the basic preclinical testing of a novel therapeutic angiogenesis strategy based on coordinated targeting of both vascular induction and maturation. Furthermore, these experiments are expected to dissect the mechanisms by which PDGF-BB signaling regulates endothelial cells and blood vessel growth and to identify alternative molecular targets, which might provide more specific therapeutic effects to modulate the dose-dependent effects of VEGF gene delivery.