Suitability of a baculoviral vector for gene delivery in a rabbit model of peripheral artery disease. Preliminary results

SIMONIN, ALEJANDRO; CASTILLO, MARTHA GIOVANNA; CUNIBERTI, LUIS; OLEA, FERNANDA DANIELA; GIMÉNEZ, CARLOS SEBASTIÁN; NÚÑEZ PEDROZO, CRISTIAN; LOCATELLI, PAOLA; CROTTOGINI, ALBERTO; BAUZÁ, MARÍA DEL ROSARIO; LÓPEZ, AYELÉN EMILCE; BELAICH, MARIANO NICOLÁS

Mar del Plata

Congreso; Reunión anual de la Sociedad Argentina de Fisiología; 2018

SAIC-SAI-SAFIS

Introduction. Peripheral artery disease (PAD) is a condition with no effective treatment. Gene therapy emerged as a possible strategy to restore blood flow to the ischemic zone. We hypothesize that baculovirus (Bv), a virus of insects not pathogenic to humans, is a suitable vector for angiogenic gene delivery in PAD. Objectives. To study the transduction efficiency (TE) in skeletal myoblasts (Msk) of rabbit hindlimb and assess the virus presence and expression profile in ischemic muscle of rabbits with PAD at different times. Methods. Msk were isolated from biopsies and cultured on a feeder layer of autologous macrophages. Then, Msk were transduced at different multiplicities of infection (MOI) with Bv encoding the green fluorescent protein (GFP) reporter gene (Bv.pCMV.GFP). TE was measured by flow cytometry. Besides, 6 rabbits with PAD were injected with 109 viral particles of Bv.pCMV.GFP and sacrificed at 3, 7 and 14 days (n=2 per time). Three additional animals injected with PBS (n=1 per time) were used as control. Injected limb biopsies were harvested and the presence of viral DNA or GFP mRNA was analyzed by RT-qPCR. GFP protein was observed by fluorescence microscopy. Results. TE was 0.3%, 9.7%, 24.2%, 43.3%, 59.4% and 74.4% for MOI 0, 25, 50, 100, 200 and 500, respectively. Viral DNA was detectable at 3 and 7 days but not at 14 days after injection. Similar profile was observed for GFP mRNA expression. GFP protein was detectable at all 3 time points post-injection. Conclusion. These preliminary results suggest that, on account of its TE and low virus persistence in the tissue, Bv is a suitable vector for angiogenic genes delivery in this model of PAD. Further safety studies to detect undesired expression in remote organs remain to be performed.