Smurf1 Silencing Using a LNA-ASOs/Lipid Nanoparticle System to Promote Bone Regeneration

Despite the great advance of bone tissue engineering in the last few years, repair of bone defects remains a major problem. Low cell engraftment and dose-dependent side effects linked to the concomitant administration of bone morphogenetic proteins (BMPs) are the main problems currently hindering the clinical use of mesenchymal stem cell (MSC)-based therapies in this field. We have managed to bypass these drawbacks by combining the silencing the Smurf1 ubiquitin ligase in MSCs with the use of a scaffold that sustainably releases low doses of BMP-2. In this system, Smurf1 silencing is achieved by using GapmeRs, a clinically safe method that avoids the use of viral vectors, facilitating its translation to the clinic. Here, we show that a single transient transfection with a small quantity of a Smurf1-specific GapmeR is able to induce a significant level of silencing of the target gene, enough to prime MSCs for osteogenic differentiation. Smurf1 silencing highly increases MSCs responsiveness to BMP-2, allowing a dramatic reduction of the dose needed to achieve the desired therapeutic effect. The combination of these primed cells with alginate scaffolds designed to sustainably and locally release low doses of BMP-2 to the defect microenvironment is able to induce the formation of a mature bone matrix both in an osteoporotic rat calvaria system and in a mouse ectopic model. Importantly, this approach also enhances osteogenic differentiation in MSCs from osteoporotic patients, characterized by a reduced bone-forming potential, even at low BMP doses, underscoring the regenerative potential of this system.ACKNOWLEDGMENTS: This research was supported by a grant from the Spanish Ministerio de Economia y competitividad (Project RTI2018-097324), a grant from the Instituto de Investigación Marqués de Valdecilla (IDIVAL, INNVAL 17/15), and Palex Medical S.A

Effective Osteogenic Priming of Mesenchymal Stem Cellsthrough LNA-ASOs-Mediated Sfrp1 Gene Silencing

Mesenchymal stem cell (MSC) transplantation has emerged as a promising approach for bone regeneration. Importantly, the beneficial effects of MSCs can be improved by modulating the expression levels of specific genes to stimulate MSC osteogenic differentiation. We have previously shown that Smurf1 silencing by using Locked Nucleic Acid-Antisense Oligonucleotides, in combination with a scaffold that sustainably releases low doses of BMP-2, was able to increase the osteogenic potential of MSCs in the presence of BMP-2 doses significantly smaller than those currently used in the clinic. This would potentially allow an important reduction in this protein in MSs-based treatments, and thus of the side effects linked to its administration. We have further improved this system by specifically targeting the Wnt pathway modulator Sfrp1. This approach not only increases MSC bone regeneration efficiency, but is also able to induce osteogenic differentiation in osteoporotic human MSCs, bypassing the need for BMP-2 induction, underscoring the regenerative potential of this system. Achieving successful osteogenesis with the sole use of LNA-ASOs, without the need of administering pro-osteogenic factors such as BMP-2, would not only reduce the cost of treatments, but would also open the possibility of targeting these LNA-ASOs specifically to MSCs in the bone marrow, allowing us to treat systemic bone loss such as that associated with osteoporosis. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Funding: This research was funded by a grant from the Spanish Ministerio de Economia y competitividad (Project RTI2018- 097324) and two grants from the Instituto de Investigación Marqués de Valdecilla-IDIVAL (PREVAL19/02 and PREVAL 20/01)

Assessing the internal uppermost crustal structure of the central pyrenees by gravity-constrained cross sections

The Pyrenees constitutes an exceptional example of an Alpine orogenic belt characterized by basement thrust sheets involving Paleozoic rocks and Mesozoic and Cenozoic cover units detached on the Triassic evaporites, the main décollement level. This work is located in the Central Pyrenees, where gravity data help to better constrain the internal architecture of the upper crust of the southern half of the Axial Zone and the northern part of the South Pyrenean Zone, a key area to understand the orogenic evolution of the chain. Previous and new gravity, petrophysical and geological data have been used to obtain the Bouguer and residual anomaly maps of the study area and six serial gravity-constrained cross sections perpendicular to the main structural trend. The residual anomaly map shows a good correlation between basement units involved in thrust sheets of the study area and gravity highs whereas negative anomalies are interpreted to correspond with Mesozoic/Cenozoic basins, Triassic evaporites, Late Variscan igneous bodies, and Ordovician gneisses. The six gravity-constrained cross sections highlight strong along-strike variations on the gravity signal due to lateral differences of the superficial and subsurface occurrence of Triassic evaporites, different geometry at depth of the Late Variscan igneous bodies outcropping in the study area, and geometric lateral variations of the basement thrust sheets and their relationship with the Mesozoic-Cenozoic units.This work was funded by projects CGL2017-84901-C2-2-P funded by MCIN/AEI/10.13039/501100011033 and “ERDF A way of making Europe”, PID2020-114273GB-C22 funded by MCIN/AEI/10.13039/501100011033 from the Spanish Ministry of Science and Innovation, and "Severo Ochoa” extraordinary grants for excellence IGME-CSIC (AECEX2021).Peer reviewe

New gravity data in the Central Pyrenees (NE Spain)

El proyecto GeoPiri3D tiene como objetivo caracterizar la geometría y la cinemática de los batolitos permo-carboníferos del Pirineo Central por medio de una combinación de campos potenciales y de datos estructurales. Durante 2018 se han adquirido 824 nuevas estaciones de gravedad y 186 muestras de roca. Los datos de gravedad se han procesado e integrado en la base de datos del IGME y se ha calculado un nuevo mapa de anomalías de Bouguer. Los dos cuerpos graníticos principales (La Maladeta y Andorra-Mount Louis) proporcionan diferentes respuestas gravimétricas. La anomalía sobre el afloramiento de granito de La Maladeta presenta pequeñas variaciones en su amplitud que se asocian con cambios de facies. Los datos petrofísicos proporcionan una densidad media de ca. 2,6 g/ cm3 que son consistentes con su composición, predominantemente granodiorítica. El afloramiento del plutón de Mount Louis-Andorra se caracteriza por un mínimo relativo, lo que sugiere una composición predominantemente granítica (aún no se tienen resultados de las medidas de densidad). Además, el mínimo se extiende en dirección NE, lo que sugiere que parte del plutón está enterrado. El mínimo relativo al SO del granito de La Maladeta probablemente está relacionado con las acumulaciones de evaporitas triásicasThe GeoPiri3D project aims to characterize the geometry and kinematics of the Permo-Carboniferous batholiths of the Central Pyrenees through a combination of potential fields and structural data. During 2018 we have acquired 824 new gravity stations and 186 rock samples. The gravity data were processed and integrated into the IGME database and a new Bouguer anomaly was calculated. The two main granite bodies (La Maladeta and Andorra-Mount Louis) provide different gravimetric responses. The anomaly on the La Maladeta granite outcrop presents a zonation with small variations in its amplitude. The petrophysical data provide an average density of ca. 2.6 g/cm3, which is consistent with its granodioritic composition. The outcrop of the Andorra-Mount Louis pluton is characterized by a relative minimum, suggesting a dominant granitic composition. In addition, the minimum extends in an oblique NE direction indicating that part of the pluton is buried. The relative minimum to the SW of La Maladeta is probably linked to accumulations of Triassic evaporite

Variaciones laterales en el basamento y cobertera de los Pirineos Centrales

La deformación Alpina en los Pirineos Centrales dio lugar a un cinturón de pliegues y láminas de cabalgamiento que involucran al basamento paleozoico y a la cobertera mesozoico-cenozoica, esta última despegada sobre las evaporitas del Triásico. La zona de estudio abarca la mitad meridional de la Zona Axial, desde el granito de la Maladeta al oeste hasta el granito de Andorra-Mont Louis al este, así como la parte más septentrional de la Zona Surpirenaica. Este trabajo tiene como objetivo analizar las variaciones laterales de la estructura del subsuelo de la zona de estudio, en base a la estructura ya establecida a partir de trabajos geológicos y geofísicos previos. Para ello se han elaborado seis cortes geológicos seriados partiendo de dichos trabajos, de cartografías geológicas previas e introduciendo datos propios; también se ha llevado a cabo la recopilación y homogenización de los datos gravimétricos existentes (base de datos SITOPO (proyecto TopoIberia) y base de datos del ICGC), y la adquisición de más de 1000 nuevos datos; finalmente se han realizado 231 medidas de densidad de todas las litologías que afloran en el área estudiada. Todo ello para elaborar los mapas de anomalías de Bouguer y anomalía gravimétrica residual de la zona, así como la modelización gravimétrica 2.5 D de los cortes geológicos realizados. Los mapas gravimétricos y los cortes seriados obtenidos permiten observar variaciones laterales relacionadas con (i) la existencia o no de rocas evaporíticas, (ii) variaciones de la geometría en profundidad de los cuerpos ígneos tardi-variscos y (iii) variaciones laterales en la geometría de las láminas de cabalgamiento alpinas que involucran al basamentoThe Alpine deformation in the Central Pyrenees generated a fold-and-thrust belt involving Paleozoic basement and decou- pled Mesozoic-Cenozoic cover units detached on the main décollement level located in the Triassic evaporites. The study area comprises the southern sector of the Axial Zone from la Maladeta granite in the west to the Andorra-Mont Louis granite to the east as well as the northernmost part of the South Pyrenean Zone. The goal of this work is to analyse lateral varia- tions of subsurface structure of the study area, which was established based on previous geological and geophysical works. With this aim, six serial geological cross sections were built using available geological maps and previous published works together with new data. More than 1000 new gravity stations were acquired and harmonized together with available data from SITOPO database (TopoIberia project) and ICGC database as well as 231 new density measurements taken from all rock types outcropping in the study area. Integration of all this information allowed us to calculate the Bouguer and residual gravity anomaly maps and to carry out 2.5 D gravity modelling along the six cross sections. Both, maps and cross sections, show lateral variations related to (i) occurrence or absence of Triassic evaporites, (ii) different geometry at depth of the Late Variscan igneous bodies and (iii) geometric lateral changes of the alpine basement-involved thrust sheet

Nuevos datos gravimétricos y geológicos de la Zona Axial y límite con la Zona Surpirenaica del Pirineo Central

El conocimiento de la geometría del subsuelo se basa en la integración de datos geológicos y geofísicos. En la Zona Axial del Pirineo Central y sector septentrional de la Zona Surpirenaica, la ausencia de perfiles de sísmica de reflexión dificulta esta labor y hace necesaria la búsqueda de otras técnicas, como la gravimetría, para inferir su estructura en profundidad. En este trabajo los equipos del IGME e ICGC han tomado 1164 nuevas estaciones gravimétricas en la zona de estudio, se han recopilado 2740 estaciones previas de las bases de datos de SITOPO (proyecto TopoIberia) e ICGC y se ha realizado su homogenización. Se ha obtenido un nuevo mapa de la anomalía de Bouguer caracterizado por un mínimo elongado de longitud de onda larga de más de -100 mGal asociado a la raíz cortical de los Pirineos. Cabe destacar la diferente respuesta gravimétrica que ofrecen los dos principales granitos de la zona de estudio, La Maladeta y Andorra-Mont Louis (menor densidad que las rocas Paleozoicas del encajante). Al SO del granito de La Maladeta aparece un mínimo relativo que se interpreta asociado a acumulaciones evaporíticas triásicas (menor densidad) en profundidad. Además se ha realizado: (i) un mapa de anomalía residual asumiendo una anomalía regional correspondiente a un polinomio de tercer grado para inferir la estructura más superficial y (ii) mapas de las derivadas vertical y horizontal de la anomalía gravimétrica residual para observar cómo varían espacialmente los valores de densidad de las rocas existentes. Este trabajo destaca el potencial de la gravimetría en el estudio de la estructura en profundidad de orógenos y cinturones de pliegues y cabalgamientos con presencia de rocas ígneas y/o evaporíticas con valores de densidad menores al de las rocas sedimentarias encajante

Towards 3D databases and harmonized 3D models at IGME-CSIC

IGME-CSIC has a highly relevant geological and geophysical database that includes a continuous digital geological cartography at 1:50000; 1:200000 and 1:1000000 scales and a fair amount of geophysical data: gravity, magnetic, well-logs in tiff and LAS format, seismic lines in tiff and SEG-Y format, borehole and petrophysical data, together with other geophysical and geological studies. Since the 2004, an important effort has been done to undertake 3D geological and geophysical modelling ranging from local studies (mineral exploration or CO2 storage sites) to regional geology for a better understanding of the subsurface structure and its geodynamic evolution as a base for other studies on natural hazards or mineral resources. These studies were ¿stand alone¿ and now IGME is designing a new strategy. It includes the available data and models harmonization (stratigraphy sequences, structural interpretations, faults distribution, seismic velocity models, spatial distribution of physical properties such as density and magnetic susceptibility, workflows, methodologies, evaluation of uncertainties, visualization, etc.) to comply with the FAIR (Findable, Accessible, Interoperable and Reusable) data standardization. In this way, the new 3D models will be easily integrated and available from the databases. This strategy includes collaboration with the Bureau de Recherches Géologiques et Minières of France (BRGM) and Laboratório Nacional de Energia e Geologia of Portugal (LNEG) in order to harmonize the Spanish geological data and models with their neighbours across national borders. The first step is being done in the framework of GeoERA projects. Plain-language Summary IGME-CSIC owns a large database that includes a highly valuable geological and geophysical data and geophysical studies containing the interpretation of some of the data of Spain (onshore and offshore) Since 2004 the authors of this work have been working in 3D geological and geophysical modelling that includes local (mineral exploration or CO2 storage sites) and regional studies. The goal is to improve our understanding of the subsurface structures and processes as a base for deepening our knowledge in how the natural hazards occur, how to improve the exploration for mineral resources, etc. These studies were made ad hoc within different projects and now IGME-CSIC is designing a workflow to harmonize these models in order to comply with the FAIR (Findable, Accessible, Interoperable and Reusable) data standardization so the models will be available to being used beyond the initial objectives that generated their creation. This strategy includes collaboration with other European institutions like the Bureau de Recherches Géologiques et Minières of France (BRGM) and Laboratório Nacional de Energia e Geologia of Portugal (LNEG) in order to harmonize the models across national borders. The first step is already being done in the framework of the GeoERA projects

Characterization of the Cerdanya Neogene Basin by combining geophysical methods: passive seismic, magnetotelluric and gravimetry.

Congreso realizado en Toledo del 28 de noviembre al 1 de diciembre de 2022.[EN] The Cerdanya Neogene basin, located in the eastern sector of the Central Pyrenees, has been studied to characterize its structure in depth. The thickness of the Neogene filling of the basin (detrital materials with some levels of lignite) has been inferred by combining different geophysical exploration methods in a NW-SE trending profile that crosses, perpendicularly, the basin in its central part: passive seismic (H/V spectral ratio method and array technique), electromagnetic methods (MT and AMT) and gravimetry. Applying the array technique, the shear wave velocity (Vs) of the basin materials has been calculated, both for the Neogene deposits and the bedrock. These data combined with the results obtained from the H/V spectral ratio method allow deriving the bedrock geometry. The electromagnetic method provides the 2D electrical resistivity model, characterizing the geoelectric properties of the basin and depicting the presence of the d'Alp-la Tet fault, an important structural element of the basin. The electrical model inversion uses as an initial model the bedrock depth obtained from the passive seismic. In the residual Bouguer anomaly modelling, a satisfactory gravimetric model is obtained when the thickness of the Neogene deposits varies between 0 and 650 meters and assigned density of 2.2 g/cm3 , supported by the low Vs obtained. The thickness of the Neogene filling presents variations that are related to the presence of normal fault with small vertical offset. The application of this methodology, based on the combination of different geophysical exploration methods, has reduced the different uncertainties inherent in each geophysical method, and aim to characterize the Cerdanya Neogene basin geometry.Este trabajo se ha financiado con el proyecto PID2020-114273GB-C22 financiado por MCIN/AEI/10.13039/501100011033 del Ministerio de Ciencia e Innovación de España.Peer reviewe

Petrophysical characterization of non-magnetic granites; density and magnetic susceptibility relationships

In this work we establish reliable correlations between density and magnetic susceptibility in three paramagnetic granites from the Pyrenees. In total, 128 sites (310 density measurements and >2600 susceptibility ones) were studied in the Mont Louis-Andorra, Maladeta and Marimanha granitic plutons covering the main range of variability of magnetic susceptibility. Regressions were calculated for every granitic body and an integrated linear function was obtained for the entire dataset: ρ (kg/m3) = 2566 (kg/m3) + 0.541κ (10−6 S.I.) (R:0.97). This relationship is only valid in the paramagnetic domain, where iron is mostly fractioned in iron-bearing phyllosilicates and the occurrence of magnetite is negligible (or at least its contribution to the bulk susceptibility). This relationship, likely different in other bodies, allows for transforming magnetic susceptibility data into density data, helping to constrain gravity modelling when density data from rock samples are scarce. Given the large amount of AMS studies worldwide, together with the quickness and cost-effectiveness of susceptibility measurements with portable devices, this methodology allows for densifying and homogenizing the petrophysical data when modelling granite rock volumes based on both magnetic and gravimetric signals.This work was financed by the projects GeoPiri3D (CGL2017-84901-C2-2-P), UKRIA4D (PID2019-104693GB-I00/CTA) and IMAGYN (PID2020-114273GB-C22) from the Spanish Ministry of Science (funded by MCIN/AEI/10.13039/501100011033 and “ERDF A way of making Europe”). This work is methodologically related also with the 3DGeoEU project financed by the European Commission under the ERANET Cofound action GeoERA (Grant No.: 731166). The GeoAp Research group from the Aragonian Government is also acknowledged. We are also in debt to the staff of the Petrophysical Laboratory (IGME, Tres Cantos) and to the Geophysics technicians (José Mª Llorente and Agustín González). The help of the Rock Magnetism Laboratory of the Centre Européen de Recherche et d’Enseignement de Géosciences de l’Environnement (CEREGE) in Aix en ProvenceMarseille is also acknowledged. P.C. acknowledges funding from PTA2017-14779-I and FJC2019- 041058-I (AEI-Spain) contracts. E.B. thanks the Geomodels Research Institute of the University of Barcelona and GGAC-2017SGR596 (Generalitat de Catalunya).Peer reviewe