Producción CyT
Congreso
Autoría
Gimenez SC
;
Castillo MG
;
Nuñes CN
;
SIMONIN, JORGE ALEJANDRO
;
Belaich MN
;
Cuniberti LA
;
Crottogini AJ
;
Olea FD
Fecha
2020
Editorial y Lugar de Edición
xxxxxx
Resumen
Información suministrada por el agente en
SIGEVA
Introduction: Peripheral artery disease (PAD) is an ailment characterized by decreased arterial blood flow to the lower limbs. Given the lack of effective treatments, angio-arteriogenesis has been proposed. We hypothesized that the administration of a baculovirus encoding mutant HIF-1α (Bv.mHIF-1α) would induce collateral vessels neoformation in the ischemic limb.Materials and methods: A. In vitro studies: 1) Transgene expression: Skeletal myoblast (Skm) isolated from rabbit adducto...
Introduction: Peripheral artery disease (PAD) is an ailment characterized by decreased arterial blood flow to the lower limbs. Given the lack of effective treatments, angio-arteriogenesis has been proposed. We hypothesized that the administration of a baculovirus encoding mutant HIF-1α (Bv.mHIF-1α) would induce collateral vessels neoformation in the ischemic limb.Materials and methods: A. In vitro studies: 1) Transgene expression: Skeletal myoblast (Skm) isolated from rabbit adductor muscle were transduced with Bv.mHIF-1 at MOI=100, and HIF-1α expression was evaluated by RT-qPCR and western blot. 2) Angiogenic potential of transduced cells: the tubulogenesis assay was performed with supernatants of SkM and SkM-mHIF-1α cultures. B. In vivo studies: 12 rabbits underwent sterile excision of the femoral artery of the left posterior limb. Seven days later, rabbits were randomized to receive in the ischemic muscle 10 injections containing 109 copies of the Bv.mHIF-1α vector (treated group, n=6) or 109 copies of Bv.null (control group, n=6). Two weeks post-treatment digital angiography was performed in both posterior limbs.Results: HIF-1α mRNA levels in SkM-mHIF-1α cells was 1000-fold higher than in non-transduced cells (p<0.05, t-test). HIF-1α protein levels were also overexpressed. Supernatants derived from SkM-mHIF-1α formed more tubular networks than those from non-transduced SkM cells (7.38±0.69 vs. 4.99± 1.01 rings/mm2, p<0.01; t-test).Finally, at 14 days post-treatment the density angiographyically visible collaterals was higher in Bv-mHIF-1α-traeatedrabbits (8.12±0.42 colaterales/cm2) than in those treated with Bv-null (6.13±1.15 colaterales/cm2; p<0,05, t-test).Conclusion: The Bv.mHIF-1α induced angiogenesis in vitro and collateral vessels neoformation in the ischemic muscle of rabbits at 14 days after treatment. Further safety and efficacy studies at longer follow up periods are needed to estimate the potential usefulness of this approach in the clinical setting.
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Palabras Clave
Terapia genicamHif1-aBaculovirusPAD