Santiago Roura - Academia.edu (original) (raw)

Papers by Santiago Roura

Cellular and Molecular Life Sciences

Huntington's disease (HD) is an incurable inherited brain disorder characterised by massive d... more Huntington's disease (HD) is an incurable inherited brain disorder characterised by massive degeneration of striatal neurons, which correlates with abnormal accumulation of misfolded mutant huntingtin (mHTT) protein. Research on HD has been hampered by the inability to study early dysfunction and progressive degeneration of human striatal neurons in vivo. To investigate human pathogenesis in a physiologically relevant context, we transplanted human pluripotent stem cell-derived neural progenitor cells (hNPCs) from control and HD patients into the striatum of new-born mice. Most hNPCs differentiated into striatal neurons that projected to their target areas and established synaptic connexions within the host basal ganglia circuitry. Remarkably, HD human striatal neurons first developed soluble forms of mHTT, which primarily targeted endoplasmic reticulum, mitochondria and nuclear membrane to cause structural alterations. Furthermore, HD human cells secreted extracellular vesicles...

European Heart Journal

Background The PERISCOPE is a first-in-human, phase I, double blind, one centre clinical trial to... more Background The PERISCOPE is a first-in-human, phase I, double blind, one centre clinical trial to test the safety of the PeriCord, an advanced therapy medicinal product (PEI18–140), for the treatment of patients with infarcted myocardial tissue. The PeriCord is a GMP-complying allogeneic engineered tissue graft consisting on a decellularised pericardial matrix colonised with umbilical cord Wharton's jelly mesenchymal stromal cells. The PeriCord implantation has shown to promote damaged tissue revascularisation, reduce infarct size, adverse remodelling and fibrosis, and ultimately improve cardiac function preclinically. Purpose To assess the safety and monitor the monocyte and cytokine response after PeriCord implantation in patients with infarcted myocardial tissue. Methods The PERISCOPE clinical trial has been approved by AEMPS and the Local Ethics Committee of our institution (Eudra-CT 2018–001964–49). Twelve patients with transmural myocardial infarction (>50% by MRI) who ...

Scientific Reports

Primary ventricular fibrillation (PVF) is a life-threatening complication of ST-segment elevation... more Primary ventricular fibrillation (PVF) is a life-threatening complication of ST-segment elevation myocardial infarction (STEMI). It is unclear what roles viral infection and/or systemic inflammation may play as underlying triggers of PVF, as a second hit in the context of acute ischaemia. Here we aimed to evaluate whether the circulating virome and inflammatory proteome were associated with PVF development in patients with STEMI. Blood samples were obtained from non-PVF and PVF STEMI patients at the time of primary PCI, and from non-STEMI healthy controls. The virome profile was analysed using VirCapSeq-VERT (Virome Capture Sequencing Platform for Vertebrate Viruses), a sequencing platform targeting viral taxa of 342,438 representative sequences, spanning all virus sequence records. The inflammatory proteome was explored with the Olink inflammation panel, using the Proximity Extension Assay technology. After analysing all viral taxa known to infect vertebrates, including humans, we ...

European Heart Journal, 2017

The regenerative therapy with induced pluripotent stem cells (iPSC)-derived cardiomyocytes for th... more The regenerative therapy with induced pluripotent stem cells (iPSC)-derived cardiomyocytes for the treatment of heart failure (HF) is eagerly awaited. But only a few studies with large animal models were performed so far. Therefore, its efficacy and safety have not been established in preclinical studies yet. We examined them by transplantation of human iPSCs-derived cardiac spheroids to adult pigs with HF. The regenerative cardiomyocytes were differentiated from human iPSC and purified by no glucose, no glutamine, and lactate-supplemented media (Tohyama and others Cell metabolism 2016). Cardiac spheroids were constituted with pure cardiomyocytes. Eight months old micro-mini pigs were used for experiments. HF models were made by cryoinjury, and iPSCs-derived cardiac spheroids with gelatin hydrogel (GH) were transplanted 4weeks later. Immunosuppressive therapies with prednisolone, mycophenolate mofetil (MMF), and tacrolimus, started at 5 days before transplantation. We defined experimental groups as iPSC group: transplantation of cardiac spheroids (1x10E8 pure cardiomyocytes) from iPSC and GH, control group: transplantation of GH. Cardiac functions were measured by cardiac magnetic resonance imaging (cMRI) that was scheduled just before cryoinjury and transplantation, and 4 weeks and 8weeks after transplantation. Cardiac telemetry leads were implanted to pigs and the frequency of arrhythmia was analyzed. Pigs were sacrificed and frozen sections of hearts were immunostained by human nuclear antigen, von Willebrand factor (vWF) and α-smooth muscle actinin (SMA) to evaluate the engraftment of iPSC-derived cardiomyocytes and neovascularization. Proportion of cardiac troponin T in purified cardiomyocytes was more than 98%. Four weeks after cryoinjury, left ventricular ejection fraction (EF) decreased to 39.9±3.1%. The blood concentration of MMF and tacrolimus were regularly monitored. Both iPSC and GH groups improved cardiac function at 4weeks after transplantation (56.0±7.9% vs 47.5±10.6%), but only iPSC group significantly improved EF in 8 weeks (56.6±6.2% vs 43.9±3.7%). Frozen sections were observed under a fluorescence microscopy. The human nuclear antigen-positive cardiomyocytes were clearly engrafted before 4 weeks, but positive area markedly decreased in 8 weeks after cell transplantation. The number of vWF and SMA positive cells were greater in iPSC group than control group. For the evaluation of arrhythmia, cardiac telemetry showed premature ventricular contractions happened around a few days after transplantation, and few happened around one month later. No pigs died of lethal ventricular arrhythmia. Co-transplantation of Human iPSC-derived cardiac spheroids with GH remarkably improved cardiac function in chronic phase. No lethal ventricular arrhythmia happened in all cases. The regenerative therapy with human iPSC is safe and effective for the treatment of HF.

JACC: Basic to Translational Science, 2016

HIGHLIGHTS MI remains a major cause of morbidity and mortality despite major treatment advances a... more HIGHLIGHTS MI remains a major cause of morbidity and mortality despite major treatment advances achieved during the past decades. Administration of an engineered myocardial graft, composed of decellularized myocardial matrix refilled with ATDPCs (EMG-ATDPC), in a porcine pre-clinical MI model, may support cardiac recovery following MI. Thirty days post-EMG-ATDPC implantation, cardiac magnetic resonance imaging and comprehensive histological analysis were performed to evaluate its impact on myocardial restoration. EMG-ATDPC resulted in better left ventricular ejection fraction, higher vessel density and neovascularization, and reduced infarct size by 68%, as well as limited fibrosis.

PloS one, 2018

Monocytes are a heterogeneous population of effector cells with key roles in tissue integrity res... more Monocytes are a heterogeneous population of effector cells with key roles in tissue integrity restoration and maintenance. Here, we explore the association of monocyte subsets and prognosis in patients with ambulatory heart failure (HF). Monocyte subsets were classified as classical (CD14++/CD16-), intermediate (CD14++/CD16+), or non-classical (CD14+/CD16++). Percentage distribution and absolute cell count were assessed in each subset, and multivariable Cox regression analyses were performed with all-cause death, HF-related hospitalization, and the composite end-point of both as dependent variables. 400 patients were consecutively included (72.8% male, age 69.4±12.2 years, 45.5% from ischemic aetiology, left ventricle ejection fraction (LVEF) 41.6% ±14.5, New York Heart Association (NYHA) class II 62.8% and III 30.8%). During a mean follow-up of 2.6±0.9 years, 107 patients died, 99 had a HF-related hospitalization and 160 suffered the composite end-point of all-cause death or HF-rel...

• MI remains a major cause of morbidity and mortality despite major treatment advances achieved d... more • MI remains a major cause of morbidity and mortality despite major treatment advances achieved during the past decades.<br>• Administration of an engineered myocardial graft, composed of decellularized myocardial matrix refilled with ATDPCs (EMG-ATDPC), in a porcine pre-clinical MI model, may support cardiac recovery following MI.<br>• Thirty days post-EMG-ATDPC implantation, cardiac magnetic resonance imaging and comprehensive histological analysis were performed to evaluate its impact on myocardial restoration.<br>• EMG-ATDPC resulted in better left ventricular ejection fraction, higher vessel density and neovascularization, and reduced infarct size by 68%, as well as limited fibrosis.<br>• Accordingly, EMG-ATDPC is ready to start the translational avenue toward phase I first-in-man clinical trials.

The existence of allogeneic cells within an individual has been demonstrated in multiple fields s... more The existence of allogeneic cells within an individual has been demonstrated in multiple fields such as hematopoieticstem cell or solid organ transplantation, non-depleted blood transfusions and the most common form which isbidirectional maternal-fetal cell trafficking, whereby cells from the fetus pass through the placental barrier. In order tographically illustrate this early natural phenomenon that initiates the journey of a child’s cells within the mother’s bloodand other tissues, we used a new procedure in microscopy imaging generating Large Scale Panoramic Pictures (LSPP).This technique can also be extended to explore a broad diversity of experimental models.

Angiogenesis, 2008

Among the molecular and cellular processes that orchestrate construction of new blood vessels, th... more Among the molecular and cellular processes that orchestrate construction of new blood vessels, the distinct contribution of circulating cells has recently become appreciated. The endothelial progenitor cell (EPC) was one of the first identified circulating cell types found to directly contribute to neo-vessels walls. Subsequently, a complex network of signals between EPCs and other circulating bone marrow-derived cell types has been described. Significant temporal and spatial heterogeneity in utilization of circulating progenitor cells exists between tumor types, complicating analysis in both animal models and in patients. A lack of standardized cell surface markers and techniques for quantitation of such rare cells has further complicated such studies. Nonetheless, levels of EPCs and other bone marrow-derived progenitors may hold prognostic significance for cancer patients or may be used in the future to guide therapy and EPCs may themselves represent a valid therapeutic target.

Stem Cell Research & Therapy, 2019

BackgroundOrthopaedic diseases are one of the major targets for regenerative medicine. In this co... more BackgroundOrthopaedic diseases are one of the major targets for regenerative medicine. In this context, Wharton’s jelly (WJ) is an alternative source to bone marrow (BM) for allogeneic transplantation since its isolation does not require an invasive procedure for cell collection and does not raise major ethical concerns. However, the osteogenic capacity of human WJ-derived multipotent mesenchymal stromal cells (MSC) remains unclear.MethodsHere, we compared the baseline osteogenic potential of MSC from WJ and BM cell sources by cytological staining, quantitative real-time PCR and proteomic analysis, and assessed chemical and biological strategies for priming undifferentiated WJ-MSC. Concretely, different inhibitors/activators of the TGFβ1-BMP2 signalling pathway as well as the secretome of differentiating BM-MSC were tested.ResultsCytochemical staining as well as gene expression and proteomic analysis revealed that osteogenic commitment was poor in WJ-MSC. However, stimulation of the...

Journal of cellular and molecular medicine, Jan 29, 2017

Idiopathic dilated cardiomyopathy (IDCM) is a frequent cause of heart transplantation. Potentiall... more Idiopathic dilated cardiomyopathy (IDCM) is a frequent cause of heart transplantation. Potentially valuable blood markers are being sought, and low-density lipoprotein receptor-related protein 1 (LRP1) has been linked to the underlying molecular basis of the disease. This study compared circulating levels of soluble LRP1 (sLRP1) in IDCM patients and healthy controls and elucidated whether sLRP1 is exported out of the myocardium through extracellular vesicles (EVs) to gain a better understanding of the pathogenesis of the disease. LRP1 α chain expression was analysed in samples collected from the left ventricles of explanted hearts using immunohistochemistry. sLRP1 concentrations were determined in platelet-free plasma by enzyme-linked immunosorbent assay. Plasma-derived EVs were extracted by size-exclusion chromatography (SEC) and characterized by nanoparticle tracking analysis and cryo-transmission electron microscopy. The distributions of vesicular (CD9, CD81) and myocardial (cave...

BioMed Research International, 2015

Over the years, cell therapy has become an exciting opportunity to treat human diseases. Early en... more Over the years, cell therapy has become an exciting opportunity to treat human diseases. Early enthusiasm using adult stem cell sources has been tempered in light of preliminary benefits in patients. Considerable efforts have been dedicated, therefore, to explore alternative cells such as those extracted from umbilical cord blood (UCB). In line, UCB banking has become a popular possibility to preserve potentially life-saving cells that are usually discarded after birth, and the number of UCB banks has grown worldwide. Thus, a brief overview on the categories of UCB banks as well as the properties, challenges, and impact of UCB-derived mesenchymal stem cells (MSCs) on the area of cardiovascular research is presented. Taken together, the experience recounted here shows that UCBMSCs are envisioned as attractive therapeutic candidates against human disorders arising and/or progressing with vascular deficit.

European Heart Journal, 2013

Journal of Biological Chemistry, 1999

Alteration of cadherin-mediated cell-cell adhesion is frequently associated to tyrosine phosphory... more Alteration of cadherin-mediated cell-cell adhesion is frequently associated to tyrosine phosphorylation of p120-and ␤-catenins. We have examined the role of this modification in these proteins in the control of ␤-catenin/E-cadherin binding using in vitro assays with recombinant proteins. Recombinant pp60 c-src efficiently phosphorylated both catenins in vitro, with stoichiometries of 1.5 and 2.0 mol of phosphate/mol of protein for ␤-catenin and p120-catenin, respectively. pp60 c-src phosphorylation had opposing effects on the affinities of ␤-catenin and p120 for the cytosolic domain of E-cadherin; it decreased (in the case of ␤-catenin) or increased (for p120) catenin/E-cadherin binding. However, a role for p120-catenin in the modulation of ␤-catenin/E-cadherin binding was not observed, since addition of phosphorylated p120-catenin did not modify the affinity of phosphorylated (or unphosphorylated) ␤-catenin for E-cadherin. The phosphorylated Tyr residues were identified as Tyr-86 and Tyr-654. Experiments using point mutants in these two residues indicated that, although Tyr-86 was a better substrate for pp60 c-src , only modification of Tyr-654 was relevant for the interaction with E-cadherin. Transient transfections of different mutants demonstrated that Tyr-654 is phosphorylated in conditions in which adherens junctions are disrupted and evidenced that binding of ␤-catenin to E-cadherin in vivo is controlled by phosphorylation of ␤-catenin Tyr-654.

Arteriosclerosis, Thrombosis, and Vascular Biology, 2013

Objective— Hypoxia disturbs vascular function by promoting extracellular matrix remodeling. Extra... more Objective— Hypoxia disturbs vascular function by promoting extracellular matrix remodeling. Extracellular matrix integrity and composition are modulated by metalloproteinases (MMPs). Our aim was to investigate the role of low-density lipoprotein receptor–related protein 1 (LRP1) in regulating MMP-9/MMP-2 activation and vascular smooth muscle cells (VSMCs) migration in response to hypoxia, and to elucidate the LRP1-signaling pathways involved in this process. Approach and Results— Western blot analysis showed that hypoxia induced a sustained phosphorylation of proline-rich tyrosine kinase 2 concomitantly with LRP1 overexpression in human VSMCs (hVSMCs). Deletion of LRP1 using small-interfering RNA technology or treatment of hVSMCs with the Src family kinase inhibitor PP2 impaired hypoxia-induced phosphorylation of proline-rich tyrosine kinase 2 levels. Coimmunoprecipitation experiments showed that the higher amounts of phosphorylation of proline-rich tyrosine kinase 2/LRP1β immunopre...

Journal of Cellular and Molecular Medicine, 2013

Introduction • More than a century of bioluminescence: description of its molecular basis • Appli... more Introduction • More than a century of bioluminescence: description of its molecular basis • Application of light-emitting proteins to bioanalysis • Imaging modalities in cell-based cardiac therapy • Luminescent expression reporters illuminate cardiac regeneration efforts • Conclusions and future perspectives

PLoS ONE, Nov 16, 2012

Stem cell therapies are promising strategies to regenerate human injured tissues, including ische... more Stem cell therapies are promising strategies to regenerate human injured tissues, including ischemic myocardium. Here, we examined the acquisition of properties associated with vascular growth by human umbilical cord blood-derived mesenchymal stem cells (UCBMSCs), and whether they promoted vascular growth in vivo. UCBMSCs were induced in endothelial cell-specific growth medium (EGM-2) acquiring new cell markers, increased Ac-LDL uptake, and migratory capacity as assessed by qRT-PCR, Western blotting, indirect immunofluorescence, and invasion assays. Angiogenic and vasculogenic potentials could be anticipated by in vitro experiments showing self organization into Matrigel-mediated cell networks, and activation of circulating angiogenic-supportive myeloid cells. In mice, following subcutaneous co-injection with Matrigel, UCBMSCs modified to co-express bioluminescent (luciferases) and fluorescent proteins were demonstrated to participate in the formation of new microvasculature connected with the host circulatory system. Response of UCBMSCs to ischemia was explored in a mouse model of acute myocardial infarction (MI). UCBMSCs transplanted using a fibrin patch survived 4 weeks post-implantation and organized into CD31 + network structures above the infarcted myocardium. MItreated animals showed a reduced infarct scar and a larger vessel-occupied area in comparison with MI-control animals. Taken together, the presented results show that UCBMSCs can be induced in vitro to acquire angiogenic and vasculogenic properties and contribute to vascular growth in vivo.