Transient gene therapy with baculoviral vector encoding mutant HIF-1α induces collateral vessels formation in rabbits with peripheral arterial disease

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

Rosario

Congreso; Reunión Científica Anual de la Sociedad Argentina de Fisiología; 2019

Introduction: Peripheral artery disease (PAD) is an ailment characterized by decreased arterial blood flow to the lower limbs. We hypothesized that the administration of a baculovirus (Bv) encoding mutant HIF-1α is safe and induces neovasculature in the ischemic limb. Methods: 1) HIF-1α expression was evaluated in transduced and non- transduce muscle cells (SkM) by RT-qPCR and western blot. 2) In vitro tubulogenesis assay was performed with supernatants of SkM and SkM-mHIF-1α cultures. 3) Rabbits underwent sterile excision of the femoral artery of the left posterior limb. Seven days later, rabbits were randomized in two protocols: A) Tracking assay: Rabbits with PAD were injected with 1E9 copies of Bv.GFP and sacrificed at 3, 7 and 14 days (n=2 per time). Two animals injected with PBS were used as controls. The presence of viral DNA or GFP mRNA in the injected limb and remote tissues biopsies were analyzed by RT-qPCR. GFP protein was observed by fluorescence microscopy. B) Therapeutic efficacy: Rabbits were randomized to receive 1E9 copies of the Bv.mHIF-1α or Bv.null (n=6 per group). Two weeks post-treatment digital angiography and immunohistology were performed in the posterior limbs. Results: HIF-1α mRNA levels in transduced SkM were 1000-fold higher than SkM (p