Papers by Maria del Rosario Bauza
Effect of intramuscular baculovirus encoding mutant hypoxia-inducible factor 1-alpha on neovasculogenesis and ischemic muscle protection in rabbits with peripheral arterial disease
Cytotherapy, 2020
BACKGROUND AIMS Peripheral arterial disease (PAD) is a progressive, disabling ailment for which n... more BACKGROUND AIMS Peripheral arterial disease (PAD) is a progressive, disabling ailment for which no effective treatment exists. Gene therapy-mediated neovascularization has emerged as a potentially useful strategy. We tested the angiogenic and arteriogenic efficacy and safety of a baculovirus (BV) encoding mutant, oxygen-resistant hypoxia-inducible factor 1-alpha (mHIF-1α), in rabbits with PAD. METHODS After assessing the transfection efficiency of the BV.mHIF-1α vector and its tubulogenesis potential in vitro, we randomized rabbits with experimental PAD to receive 1 × 109 copies of BV.mHIF-1α or BV.null (n = 6 per group) 7 days after surgery. Two weeks post-treatment, collateralization (digital angiography) and capillary and arteriolar densities (immunohistochemistry) were measured in the posterior limbs. Ischemic damage was evaluated in adductor and gastrocnemius muscle samples. Tracking of viral DNA in injected zones and remote tissues at different time points was performed in additional rabbits using a BV encoding GFP. RESULTS Angiographically visible collaterals were more numerous in BV.mHIF-1α-treated rabbits (8.12 ± 0.42 vs 6.13 ± 1.15 collaterals/cm2, P < 0.05). The same occurred with arteriolar (27.9 ± 7.0 vs 15.3 ± 4.0 arterioles/mm2) and capillary (341.8 ± 109.9 vs 208.8 ± 87.7 capillaries/mm2, P < 0.05) densities. BV.mHIF-1α-treated rabbits displayed less ischemic muscle damage than BV.null-treated animals. Viral DNA and GFP mRNA were detectable only at 3 and 7 days after injection in hind limbs. Neither the virus nor GFP mRNA was detected in remote tissues. CONCLUSIONS In rabbits with PAD, BV.mHIF-1α induced neovascularization and reduced ischemic damage, exhibiting a good safety profile at 14 days post-treatment. Complementary studies to evaluate its potential usefulness in the clinic are needed.
Artificial Cells, Nanomedicine, and Biotechnology, 2018
Crottogini (2018) Effect of poly (l-lactic acid) scaffolds seeded with aligned diaphragmatic myob... more Crottogini (2018) Effect of poly (l-lactic acid) scaffolds seeded with aligned diaphragmatic myoblasts overexpressing connexin-43 on infarct size and ventricular function in sheep with acute coronary occlusion, Artificial Cells, Nanomedicine, and Biotechnology, 46:sup3, S717-S724,
Cytotechnology, Jan 15, 2017
Diaphragmatic myoblasts (DMs) are precursors of type-1 muscle cells displaying high exhaustion th... more Diaphragmatic myoblasts (DMs) are precursors of type-1 muscle cells displaying high exhaustion threshold on account that they contract and relax 20 times/min over a lifespan, making them potentially useful in cardiac regeneration strategies. Besides, it has been shown that biomaterials for stem cell delivery improve cell retention and viability in the target organ. In the present study, we aimed at developing a novel approach based on the use of poly (L-lactic acid) (PLLA) scaffolds seeded with DMs overexpressing connexin-43 (cx43), a gap junction protein that promotes inter-cell connectivity. DMs isolated from ovine diaphragm biopsies were characterized by immunohistochemistry and ability to differentiate into myotubes (MTs) and transduced with a lentiviral vector encoding cx43. After confirming cx43 expression (RT-qPCR and Western blot) and its effect on inter-cell connectivity (fluorescence recovery after photobleaching), DMs were grown on fiber-aligned or random PLLA scaffolds. ...
Gene therapy
Journal of Cardiovascular Pharmacology, 2021
Adult mammalian cardiomyocytes show scarce division ability, which makes the heart ineffective in... more Adult mammalian cardiomyocytes show scarce division ability, which makes the heart ineffective in replacing lost contractile cells after ischemic cardiomyopathy. In the past decades, there have been increasing efforts in the search for novel strategies to regenerate the injured myocardium. Among them, gene therapy is one of the most promising ones, based on recent and emerging studies that support the fact that functional cardiomyocyte regeneration can be accomplished by the stimulation and enhancement of the endogenous ability of these cells to achieve cell division. This capacity can be targeted by stimulating several molecules, such as cell cycle regulators, noncoding RNAs, transcription, and metabolic factors. Therefore, the proposed target, together with the selection of the vector used, administration route, and the experimental animal model used in the development of the therapy would determine the success in the clinical field.
American Journal of Physiology-Heart and Circulatory Physiology, 2020
The adult mammalian cardiomyocyte has a very limited capacity to reenter the cell cycle and advan... more The adult mammalian cardiomyocyte has a very limited capacity to reenter the cell cycle and advance into mitosis. Therefore, diseases characterized by lost contractile tissue usually evolve into myocardial remodeling and heart failure. Analyzing the cardiac transcriptome at different developmental stages in a large mammal closer to the human than laboratory rodents may serve to disclose positive and negative cardiomyocyte cell cycle regulators potentially targetable to induce cardiac regeneration in the clinical setting. Thus we aimed at characterizing the transcriptomic profiles of the early fetal, late fetal, and adult sheep heart by employing RNA-seq technique and bioinformatic analysis to detect protein-encoding genes that in some of the stages were turned off, turned on, or differentially expressed. Genes earlier proposed as positive cell cycle regulators such as cyclin A, cdk2, meis2, meis3, and PCNA showed higher expression in fetal hearts and lower in AH, as expected. In con...
High-dose intramyocardial HMGB1 induces long-term cardioprotection in sheep with myocardial infarction
Drug Delivery and Translational Research, 2019
In rodents with acute myocardial infarction (AMI), high mobility group box 1 (HMGB1) injection ha... more In rodents with acute myocardial infarction (AMI), high mobility group box 1 (HMGB1) injection has produced controversial results. Given the lack of data in large mammals, we searched the dose that would promote angiogenesis and expression of specific regenerative genes in sheep with AMI (protocol 1) and, subsequently, use this dose to study long-term effects on infarct size and left ventricular (LV) function (protocol 2). Protocol 1: Sheep with AMI received 250 μg (high-dose, n = 7), 25 μg (low-dose, n = 7) HMGB1, or PBS (placebo, n = 7) in 10 intramyocardial injections (0.2 ml each) in the peri-infarct area. Seven days later, only the high-HMGB1-dose group exhibited higher microvascular densities, Ki67-positive cardiomyocytes, and overexpression of VEGF, Ckit, Tbx20, Nkx2.5, and Gata4. Protocol 2: Sheep with AMI received HMGB1 250 μg (n = 6) or PBS (n = 6). At 60 days, HMGB1-treated sheep showed smaller infarcts (8.5 ± 2.11 vs. 12.2 ± 1.97% LV area, P < 0.05, ANOVA-Bonferroni) and higher microvascular density (capillaries, 1798 ± 252 vs. 1266 ± 250/mm2; arterioles, 18.3 ± 3.9 vs. 11.7 ± 2.2/mm2; both P < 0.01). Echocardiographic LV ejection fraction, circumferential shortening, and wall thickening increased from day 3 to 60 with HMGB1 (all P < 0.05). Conclusion: in ovine AMI, high-dose HMGB1 induces angio-arteriogenesis, reduces infarct size, and improves LV function at 2 months post-treatment.
Cardiovascular care-Biology II 937 tivated protein-1 family of trascription factors, is emerging ... more Cardiovascular care-Biology II 937 tivated protein-1 family of trascription factors, is emerging as a major gatekeeper against oxidative stress. However, its role in diabetic cardiomyopathy remains unknown. The present study was designed to investigate whether cardiac specific overexpression of JunD protects against oxidative stress and myocardial dysfunction in the diabetic heart. Methods: Diabetes was induced in C57BL/6 wild type (WT) and cardiac specific JunD transgenic (α-MHC-JunDtg) male mice by streptozotocin (4-6 months old; n=10 per group). Control mice of both groups received citrate buffer only. Left ventricular (LV) function was assessed by high resolution Micro-Ultrasound System (Vevo 2100, Visualsonics) in diabetic and control mice after four weeks of diabetes. Then, mice were euthanized and hearts collected to determine cardiac JunD mRNA and protein expression, superoxide anion (O2-) production by ESR spectroscopy and expression of pro-oxidant and scavenging enzymes by Western blot. Results: LV dysfunction with reduced ejection fraction and fractional shortening was observed in WT diabetic mice. They also showed a significant downregulation of JunD cardiac expression. Accordingly, the expression of free radicals scavangers SOD1 and ALDH2 was reduced. By contrast, Nox2 and Nox4 subunits of NADPH oxidase were upregulated resulting in higher generation of ROS from the hearts of diabetic as compared to control WT mice. Interestingly enough, diabetic α-MHC-JunDtg mice did not show any of these hyperglycemia-induced detrimental changes and LV function was preserved. Conclusions: The present study shows that cardiac JunD protects against diabetes-induced myocardial dysfunction and may represent a novel target to prevent hyperglycemia-induced, ROS-driven cardiac damage Acknowledgement/Funding: This study was supported by grants from Swedish Heart-Lung Foundation, EFSD/Novo Nordisk Award and Vetenskaprådet.
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Papers by Maria del Rosario Bauza