Ruiz-Villalba, A. (Adrián)
- Publications
- item.page.relationships.isContributorAdvisorOfPublication
- item.page.relationships.isContributorOfPublication
6 results
Search Results
Now showing 1 - 6 of 6
- Functional modulation of atrio-ventricular conduction by enhanced late sodium current and calcium-dependent mechanisms in Scn5a1798insD/þ mice(2020) Remme, C.A. (Carol Ann); Rajamani, S. (Sridharan); Rivaud, M.R. (Mathilde R.); van-Veen, T.A.B. (Toon A.B.); van-der-Made, I. (Ingeborg); Beekman, L. (Leander); Ruiz-Villalba, A. (Adrián); Belardinelli, L. (Luiz); Basso, C. (Cristina); Marchal, G.A. (Gerard A.); Wolswinkel, R. (Rianne); Creemers, E.E. (Ester E.); Baartscheer, A. (Antonius); Jansen, J.A. (John A.); Bezzina, C.R. (Connie R.); Thiene, G. (Gaetano)Aims SCN5A mutations are associated with arrhythmia syndromes, including Brugada syndrome, long QT syndrome type 3 (LQT3), and cardiac conduction disease. Long QT syndrome type 3 patients display atrio-ventricular (AV) conduction slowing which may contribute to arrhythmogenesis. We here investigated the as yet unknown underlying mechanisms. Methods and results We assessed electrophysiological and molecular alterations underlying AV-conduction abnormalities in mice carrying the Scn5a1798insD/þ mutation. Langendorff-perfused Scn5a1798insD/þ hearts showed prolonged AV-conduction compared to wild type (WT) without changes in atrial and His-ventricular (HV) conduction. The late sodium current (INa,L) inhibitor ranolazine (RAN) normalized AV-conduction in Scn5a1798insD/þ mice, likely by preventing the mutation-induced increase in intracellular sodium ([Naþ]i ) and calcium ([Ca2þ]i ) concentrations. Indeed, further enhancement of [Naþ]i and [Ca2þ]i by the Naþ/Kþ-ATPase inhibitor ouabain caused excessive increase in AV-conduction time in Scn5a1798insD/þ hearts. Scn5a1798insD/þ mice from the 129P2 strain displayed more severe AV-conduction abnormalities than FVB/N-Scn5a1798insD/þ mice, in line with their larger mutation-induced INa,L. Transverse aortic constriction (TAC) caused excessive prolongation of AV-conduction in FVB/N-Scn5a1798insD/þ mice (while HV-intervals remained unchanged), which was prevented by chronic RAN treatment. Scn5a1798insD/þTAC hearts showed decreased mRNA levels of conduction genes in the AV-nodal region, but no structural changes in the AV-node or His bundle. In Scn5a1798insD/þ-TAC mice deficient for the transcription factor Nfatc2 (effector of the calcium-calcineurin pathway), AV-conduction and conduction gene expression were restored to WT levels. Conclusions Our findings indicate a detrimental role for enhanced INa,L and consequent calcium dysregulation on AVconduction in Scn5a1798insD/þ mice, providing evidence for a functional mechanism underlying AV-conduction disturbances secondary to gain-of-function SCN5A mutations.
- Delivery of cardiovascular progenitors with biomimetic microcarriers reduces adverse ventricular remodeling in a rat model of chronic myocardial infarction(Elsevier, 2021) Hernández, S.C. (Silvia C.); Saludas-Echauri, L. (Laura); Blanco-Prieto, M.J. (María José); Palacios, I. (Itziar); Ruiz-Villalba, A. (Adrián); Garbayo, E. (Elisa); Roncal, C. (Carmen); Prosper-Cardoso, F. (Felipe); Pelacho, B. (Beatriz); Abizanda-Sarasa, G. (Gloria); Sanchez, B. (Belén)Despite tremendous progress in cell-based therapies for heart repair, many challenges still exist. To en- hance the therapeutic potential of cell therapy one approach is the combination of cells with biomaterial delivery vehicles. Here, we developed a biomimetic and biodegradable micro-platform based on polymeric microparticles (MPs) capable of maximizing the therapeutic potential of cardiac progenitor cells (CPCs) and explored its efficacy in a rat model of chronic myocardial infarction. The transplantation of CPCs adhered to MPs within the infarcted myocardial microenvironment improved the long-term engraftment of trans- planted cells for up to one month. Furthermore, the enhancement of cardiac cellular retention correlated with an increase in functional recovery. In consonance, better tissue remodeling and vasculogenesis were observed in the animals treated with cells attached to MPs, which presented smaller infarct size, thicker right ventricular free wall, fewer deposition of periostin and greater density of vessels than animals treated with CPCs alone. Finally, we were able to show that part of this beneficial effect was mediated by CPC- derived extracellular vesicles (EVs). Taken together, these findings indicate that the biomimetic microcarri- ers support stem cell survival and increase cardiac function in chronic myocardial infarction through mod- ulation of cardiac remodeling, vasculogenesis and CPCs-EVs mediated therapeutic effects. The biomimetic microcarriers provide a solution for biomaterial-assisted CPC delivery to the heart.
- Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative study(Elsevier, 2022) Hernández, S.C. (Silvia C.); Saludas-Echauri, L. (Laura); Blanco-Prieto, M.J. (María José); Vader, P. (Pieter); Ruiz-Villalba, A. (Adrián); Garbayo, E. (Elisa); Prosper-Cardoso, F. (Felipe)Since the discovery of the beneficial therapeutical effects of extracellular vesicles (EVs), these agents have been attracting great interest as next-generation therapies. EVs are nanosized membrane bodies secreted by all types of cells that mediate cell–cell communication. Although the classification of different subpopulations of EVs can be complex, they are broadly divided into microvesicles and exosomes based on their biogenesis and in large and small EVs based on their size. As this is an emerging field, current investigations are focused on basic aspects such as the more convenient method for EV isolation. In the present paper, we used cardiac progenitor cells (CPCs) to study and compare different cell culture conditions for EV isolation as well as two of the most commonly employed purification methods: ultracentrifugation (UC) and size-exclusion chromatography (SEC). Large and small EVs were separately analysed. We found that serum starvation of cells during the EV collecting period led to a dramatic decrease in EV secretion and major cell death. Regarding the isolation method, our findings suggest that UC and SEC gave similar EV recovery rates. Separation of large and small EV-enriched subpopulations was efficiently achieved with both purification protocols although certain difference in sample heterogeneity was observed. Noteworthy, while calnexin was abundant in large EVs, ALIX and CD63 were mainly found in small EVs. Finally, when the functionality of EVs was assessed on primary culture of adult murine cardiac fibroblasts, we found that EVs were taken up by these cells, which resulted in a pronounced reduction in the proliferative and migratory capacity of the cells. Specifically, a tendency towards a larger effect of SEC-related EVs was observed. No differences could be found between large and small EVs. Altogether, these results contribute to establish the basis for the use of EVs as therapeutic platforms, in particular in regenerative fields.
- Generation of heart and vascular system in rodents by blastocyst complementation(2023) Ullate-Agote, A. (Asier); Abizanda-Sarasa, G. (Gloria María); Aranguren-López, X. (Xabier); San-Martín-Uriz, P. (Patxi); Barreda, C. (Carolina); Coppiello, G. (Giulia); Larequi-Ardanaz, E. (Eduardo); Carvajal-Vergara, X. (Xonia); Pelacho-Samper, B. (Beatriz); Ruiz-Villalba, A. (Adrián); Mazo, M. (Manuel); Arellano-Viera, E. (Estíbaliz); Pérez-Pomares, J.M. (José María); Linares, J. (Javier); Pogontke, C. (Cristina); Iglesias, E. (Elena); Prosper-Cardoso, F. (Felipe); Moya-Jódar, M. (Marta); Barlabé-Ginesta, P. (Paula)Generating organs from stem cells through blastocyst complementation is a promising approach to meet the clinical need for transplants. In order to generate rejection-free organs, complementation of both parenchymal and vascular cells must be achieved, as endothelial cells play a key role in graft rejection. Here, we used a lineage-specific cell ablation system to produce mouse embryos unable to form both the cardiac and vascular systems. By mouse intraspecies blastocyst complementation, we rescued heart and vascular system development separately and in combination, obtaining complemented hearts with cardiomyocytes and endothelial cells of exogenous origin. Complemented chimeras were viable and reached adult stage, showing normal cardiac function and no signs of histopathological defects in the heart. Furthermore, we implemented the cell ablation system for rat-to-mouse blastocyst complementation, obtaining xenogeneic hearts whose cardiomyocytes were completely of rat origin. These results represent an advance in the experimentation towards the invivo generation of transplantable organs.
- Development of an injectable alginate-collagen hydrogel for cardiac delivery of extracellular vesicles(2022) Saludas-Echauri, L. (Laura); Blanco-Prieto, M.J. (María José); Echanove-González-de-Anleo, M. (Miguel); Ruiz-Villalba, A. (Adrián); Garbayo, E. (Elisa); Gil-Cabrerizo, P. (Paula); Prosper-Cardoso, F. (Felipe); Abizanda-Sarasa, G. (Gloria)Extracellular vesicles (EVs) are nanosized pArtículos with attractive therapeutic potential for cardiac repair. However, low retention and stability after systemic administration limit their clinical translation. As an alternative, the combination of EVs with biomaterial-based hydrogels (HGs) is being investigated to increase their exposure in the myocardium and achieve an optimal therapeutic effect. In this study, we developed and characterized a novel injectable in-situ forming HG based on alginate and collagen as a cardiac delivery vehicle for EVs. Different concentrations of alginate and collagen crosslinked with calcium gluconate were tested. Based on injectability studies, 1% alginate, 0.5 mg/mL collagen and 0.25% calcium gluconate HG was selected as the idoneous combination for cardiac administration using catheter-based systems. Rheological examination revealed that the HG possessed an internal gel structure, weak mechanical properties and low viscosity, facilitating an easy administration. In addition, EVs were successfully incorporated and homogeneously distributed in the HG. After administration in a rat model of myocardial infarction, the HG showed long-term retention in the heart and allowed for a sustained release of EVs for at least 7 days. Thus, the combination of HGs and EVs represents a promising therapeutic strategy for myocardial repair. Besides EVs delivery, the developed HG could represent a useful platform for cardiac delivery of multiple therapeutic agents.
- Extracellular vesicle-based therapeutics for heart repair(2021) Saludas-Echauri, L. (Laura); Blanco-Prieto, M.J. (María José); Oliveira, C.C. (Claudia C.); Ruiz-Villalba, A. (Adrián); Garbayo, E; Roncal, C. (Carmen); Prosper-Cardoso, F. (Felipe)Extracellular vesicles (EVs) are constituted by a group of heterogeneous membrane vesicles secreted by most cell types that play a crucial role in cell-cell communication. In recent years, EVs have been postulated as a relevant novel therapeutic option for cardiovascular diseases, including myocardial infarction (MI), partially outperforming cell therapy. EVs may present several desirable features, such as no tumorigenicity, low immunogenic potential, high stability, and fine cardiac reparative efficacy. Furthermore, the natural origin of EVs makes them exceptional vehicles for drug delivery. EVs may overcome many of the limitations associated with current drug delivery systems (DDS), as they can travel long distances in body fluids, cross biological barriers, and deliver their cargo to recipient cells, among others. Here, we provide an overview of the most recent discoveries regarding the therapeutic potential of EVs for addressing cardiac damage after MI. In addition, we review the use of bioengineered EVs for targeted cardiac delivery and present some recent advances for exploiting EVs as DDS. Finally, we also discuss some of the most crucial aspects that should be addressed before a widespread translation to the clinical arena.