Search for Reliable Circulating Biomarkers to Predict Carotid Plaque Vulnerability

Altres ajuts: This research was funded by grants 158/U/2017 from Fundacio La Marato TV3 and from the Instituto de Salud Carlos III (co-financed by the European Regional Development Fund). P.C.R. and E.J.X. are members of RETICS INVICTUS PLUS . S.B. and N.P. aremembers of the Quality Research Group 2017-SGR-1149 from Generalitat de Catalunya, and they are members of the Spanish Atherosclerosis Society Vascular Biology Group.Atherosclerosis is responsible for 20% of ischemic strokes, and the plaques from the internal carotid artery the most frequently involved. Lipoproteins play a key role in carotid atherosclerosis since lipid accumulation contributes to plaque progression and chronic inflammation, both factors leading to plaque vulnerability. Carotid revascularization to prevent future vascular events is reasonable in some patients with high-grade carotid stenosis. However, the degree of stenosis alone is not sufficient to decide upon the best clinical management in some situations. In this context, it is essential to further characterize plaque vulnerability, according to specific characteristics (lipid-rich core, fibrous cap thinning, intraplaque hemorrhage). Although these features can be partly detected by imaging techniques, identifying carotid plaque vulnerability is still challenging. Therefore, the study of circulating biomarkers could provide adjunctive criteria to predict the risk of atherothrombotic stroke. In this regard, several molecules have been found altered, but reliable biomarkers have not been clearly established yet. The current review discusses the concept of vulnerable carotid plaque, and collects existing information about putative circulating biomarkers, being particularly focused on lipid-related and inflammatory molecules

Sex-Dependent End-of-Life Mental and Vascular Scenarios for Compensatory Mechanisms in Mice with Normal and AD-Neurodegenerative Aging

Life expectancy decreases with aging, with cardiovascular, mental health, and neurodegenerative disorders strongly contributing to the total disability-adjusted life years. Interestingly, the morbidity/mortality paradox points to females having a worse healthy life expectancy. Since bidirectional interactions between cardiovascular and Alzheimer's diseases (AD) have been reported, the study of this emerging field is promising. In the present work, we further explored the cardiovascular-brain interactions in mice survivors of two cohorts of non-transgenic and 3xTg-AD mice, including both sexes, to investigate the frailty/survival through their life span. Survival, monitored from birth, showed exceptionally worse mortality rates in females than males, independently of the genotype. This mortality selection provided a "survivors" cohort that could unveil brain-cardiovascular interaction mechanisms relevant for normal and neurodegenerative aging processes restricted to long-lived animals. The results show sex-dependent distinct physical (worse in 3xTg-AD males), neuropsychiatric-like and cognitive phenotypes (worse in 3xTg-AD females), and hypothalamic-pituitary-adrenal (HPA) axis activation (higher in females), with higher cerebral blood flow and improved cardiovascular phenotype in 3xTg-AD female mice survivors. The present study provides an experimental scenario to study the suggested potential compensatory hemodynamic mechanisms in end-of-life dementia, which is sex-dependent and can be a target for pharmacological and non-pharmacological interventions

Clinical and radiological characteristics and outcome of wake-up intracerebral hemorrhage

There is little information on the characteristics of patients with wake-up intracerebral hemorrhage (WU-ICH). We aimed to evaluate frequency and relevant differences between WU-ICH and while-awake (WA) ICH patients. This is a retrospective study of a prospective database of consecutive patients with spontaneous ICH, who were classified as WU-ICH, WA-ICH or UO-ICH (unclear onset). We collected demographic, clinical and radiological data, prognostic and therapeutic variables, and outcome [(neurological deterioration, mortality, functional outcome (favorable when modified Rankin scale score 0-2)]. From a total of 466 patients, 98 (25.8%) were classified as UO-ICH according to the type of onset and therefore excluded. We studied 368 patients (mean age 73.9 ± 13.8, 51.4% men), and compared 95 (25.8%) WU-ICH with 273 (74.2%) WA-ICH. Patients from the WU-ICH group were significantly older than WA-ICH (76.9 ± 14.3 vs 72.8 ± 13.6, p = 0.01) but the vascular risk factors were similar. Compared to the WA-ICH group, patients from the WU-ICH group had a lower GCS score or a higher NIHSS score and a higher ICH score, and were less often admitted to a stroke unit or intensive care unit. There were no differences between groups in location, volume, rate of hematoma growth, frequency of intraventricular hemorrhage and outcome. One in five patients with spontaneous ICH are WU-ICH patients. Other than age, there are no relevant differences between WU and WA groups. Although WU-ICH is associated with worse prognostic markers vital and functional outcome is similar to WA-ICH patients

The H-ATOMIC Criteria for the Etiologic Classification of Patients with Intracerebral Hemorrhage

Background and Purpose There are no generally accepted criteria for the etiologic classification of intracerebral hemorrhage (ICH). For this reason, we have developed a set of etiologic criteria and have applied them to a large number of patients to determine their utility. Methods The H-ATOMIC classification includes 7 etiologic categories: Hypertension, cerebral Amyloid angiopathy, Tumour, Oral anticoagulants, vascular Malformation, Infrequent causes and Cryptogenic. For each category, the etiology is scored with three degrees of certainty: Possible(3), Probable(2) and Definite(1). Our aim was to perform a basic study consisting of neuroimaging, blood tests, and CT-angio when a numerical score (SICH) suggested an underlying structural abnormality. Combinations of >1 etiologic category for an individual patient were acceptable. The criteria were evaluated in a multicenter and prospective study of consecutive patients with spontaneous ICH. Results Our study included 439 patients (age 70.8 ± 14.5 years; 61.3% were men). A definite etiology was achieved in 176 (40.1% of the patients: Hypertension 28.2%, cerebral Amyloid angiopathy 0.2%, Tumour 0.2%, Oral anticoagulants 2.2%, vascular Malformation 4.5%, Infrequent causes 4.5%). A total of 7 patients (1.6%) were cryptogenic. In the remaining 58.3% of the patients, ICH was attributable to a single (n = 56, 12.7%) or the combination of 2 (n = 200, 45.5%) possible/probable etiologies. The most frequent combinations of etiologies involved possible hypertension with possible CAA (H3A3, n = 38) or with probable CAA (H3A2, n = 29), and probable hypertension with probable OA (H2O2, n = 27). The most frequent category with any degree of certainty was hypertension (H1+2+3 = 80.6%) followed by cerebral amyloid angiopathy (A1+2+3 = 30.9%). Conclusions According to our etiologic criteria, only about 40% patients received a definite diagnosis, while in the remaining patients ICH was attributable to a single possible/probable etiology or to more than one possible/probable etiology. The use of these criteria would likely help in the management of patients with ICH.This work was supported by Ministery of Health-Instituto de Salud Carlos III: RETICS (Redes temáticas de Investigación Cooperativa) INVICTUS RD012/0014 (JM-F, PC-R, AM-D, LP-S, RD-M), FEDER (Fondo Europeo de Desarrollo Regional)

A first update on mapping the human genetic architecture of COVID-19

peer reviewe

Caracterización de la proliferación celular en las zonas subventriculares y de la evolución del infarto mediante espectroscopía de resonancia magnética en un modelo preclínico de isquemia cerebral

El ictus es una patología con una elevada incidencia en la población y los pacientes afectados sufren secuelas que les causan algún tipo de discapacidad. A nivel terapéutico, la posibilidad de incidir sobre la evolución de la enfermedad es a día de hoy aún escasa. Se ha descrito que la neurogénesis, la generación de nuevas neuronas con capacidad para migrar a la zona afectada, aumenta en las zonas subventriculares (ZSVs) y el hipocampo tras un infarto cerebral, en modelos animales y en humanos. Ante este panorama, los estudios dedicados a la comprensión de los mecanismos fisiopatológicos de la neurorreparación, así como el diseño de terapias que incidan sobre dichos mecanismos pueden ser de interés para la mejora de la calidad de vida de los pacientes tras sufrir un ictus. Actualmente se están desarrollando diversas terapias dedicadas a estimular la neurogénesis endógena, la mayoría a nivel preclínico. Una de las necesidades actuales para poder realizar ensayos clínicos sobre neurogénesis en el futuro, sería disponer de biomarcadores que permitieran detectar y monitorizar la neurogénesis de forma no invasiva. Ello nos permitiría conocer las variables asociadas a una mejor o peor capacidad de reparación, observar cómo varía esta capacidad a lo largo del tiempo, cómo varía con la administración de diversos tratamientos y cómo se relaciona esta actividad con el resultado funcional a corto y largo plazo tras un infarto cerebral. La espectroscopía de resonancia magnética (ERM) es una técnica que permite la evaluación bioquímica, in vivo, del tejido de interés de forma no invasiva, incluso permite la obtención de imágenes moleculares de las distribución de un metabolito en el cerebro. No obstante, antes de poder trasladar estos conocimientos al campo clínico, es necesario validar dichos biomarcadores espectroscópicos en modelos animales que permitan correlacionarlos con técnicas inmunohistoquímicas o de microscopía (las actuales técnicas de referencia para la determinación de la neurogénesis). En esta tesis se han desarrollado protocolos de ERM para la caracterización del patrón espectral tanto para las ZSVs como para la zona de infarto en un modelo oclusión transitoria de la arteria cerebral media en rata. Además, para una caracterización metabolómica más precisa, se aplicaron otras técnicas de resonancia que podían permitir una mejor resolución espectral, como el análisis de biopsias de tejido mediante resonancia magnética de alto campo High Resolution Magic Angle Spinning (HRMAS) previo sacrificio de los animales por irradiación con microondas focalizadas (FMW) para detener el metabolismo post mortem. También se utilizaron herramientas informáticas de reconocimiento de patrones para la determinación de los metabolitos con características diferenciales de las ZSVs y la evolución del infarto en base al análisis simultáneo de varios metabolitos. Asimismo, se realizaron pruebas con el fármaco Citicolina para verificar si la técnica permitía detectar cambios en el patrón espectroscópico en las regiones de interés debido a la administración del fármaco. El uso de estas técnicas ha permitido determinar posibles biomarcadores relacionados con apoptosis en la zona de infarto y proliferación en la ZSVs así como la valoración de los efectos de la Citicolina en el patrón metabólico correlacionando los hallazgos espectroscópicos con análisis inmunohistoquímico Los resultados obtenidos ponen de relieve el potencial de la ERM en el análisis bioquímico para determinar biomarcadores con potencial traslacional a la práctica clínica aplicables a futuros estudios en el campo del ictus.Stroke is a disease with a high incidence in the population and the affected patients have sequelae that cause disabilities. From a therapeutic point of view, the possibility of treating the evolution of the disease is nowadays still limited. Neurogenesis, the generation of new neurons with the capacity of migrating to the injured region, has been described to increase in the subventricular zones (SVZs) and hippocampus post-stroke, in animal models and also in humans. That is why studies dedicated to the understanding of the neurorepair physiopathology and the design of new therapies that could influence their mechanisms could be of interest for improving the quality of life of the patients after suffering a stroke. New therapies to promote endogenous neurogenesis enhancement are currently being developed, most of them in preclinical studies. One of the needs to develop neurogenesis clinical trials in the future is to determine biomarkers that would detect and monitor neurogenesis noninvasively. This could be useful to identify associated variables with a better or worse repair capacity, how this capacity changes over time and with pharmacological treatment and how this activity correlates with functional outcome in early and late stages post-stroke. Magnetic resonance spectroscopy (MRS) is a noninvasive technique that allows metabolic profile analysis of the tissue of interest in vivo. Moreover, it allows to perform metabolic imaging of the metabolite distribution through the brain. However, before using spectroscopic biomarkers in the clinical field they must be validated in animal models so that they can be correlated with immunohistochemistry or microscopy (the actual gold standard techniques to determine neurogenesis). In this work MRS protocols were developed to characterize the spectroscopic pattern for the SVZs and also for the stroke zone in a transient middle cerebral artery occlusion rat model. Also, for a more accurate metabolomic characterization with better spectral resolution, other magnetic resonance techniques were applied like biopsy analysis with High Resolution Magic Angle Spinning (HRMAS) using prior focused microwave irradiation (FMW) animal sacrifice in order to arrest post-mortem metabolism. In addition, pattern recognition software was applied to determine feature differences between SVZs or stroke evolution using simultaneous analysis of several metabolites. MRS experiments were performed using a therapeutic agent (Citicoline) to determine if changes in the spectroscopic pattern due to drug administration could be detected. The combination of these techniques has allowed to determine potential biomarkers associated to apoptosis in the stroke zone and proliferation in the SVZs and also to determine Citicoline effects in the metabolic pattern correlating the spectroscopic findings with immunohistochemistry analysis. The results highlight that MRS is a useful technique to perform biochemical analysis to determine biomarkers with translational potential to the clinical practice that could be used for future studies in stroke

Search for Reliable Circulating Biomarkers to Predict Carotid Plaque Vulnerability

Altres ajuts: This research was funded by grants 158/U/2017 from Fundacio La Marato TV3 and from the Instituto de Salud Carlos III (co-financed by the European Regional Development Fund). P.C.R. and E.J.X. are members of RETICS INVICTUS PLUS . S.B. and N.P. aremembers of the Quality Research Group 2017-SGR-1149 from Generalitat de Catalunya, and they are members of the Spanish Atherosclerosis Society Vascular Biology Group.Atherosclerosis is responsible for 20% of ischemic strokes, and the plaques from the internal carotid artery the most frequently involved. Lipoproteins play a key role in carotid atherosclerosis since lipid accumulation contributes to plaque progression and chronic inflammation, both factors leading to plaque vulnerability. Carotid revascularization to prevent future vascular events is reasonable in some patients with high-grade carotid stenosis. However, the degree of stenosis alone is not sufficient to decide upon the best clinical management in some situations. In this context, it is essential to further characterize plaque vulnerability, according to specific characteristics (lipid-rich core, fibrous cap thinning, intraplaque hemorrhage). Although these features can be partly detected by imaging techniques, identifying carotid plaque vulnerability is still challenging. Therefore, the study of circulating biomarkers could provide adjunctive criteria to predict the risk of atherothrombotic stroke. In this regard, several molecules have been found altered, but reliable biomarkers have not been clearly established yet. The current review discusses the concept of vulnerable carotid plaque, and collects existing information about putative circulating biomarkers, being particularly focused on lipid-related and inflammatory molecules

Electronegative LDL Promotes Inflammation and Triglyceride Accumulation in Macrophages

Altres ajuts: This research was funded by grants 158/U/2017 from Fundacio La Marato TV3.Electronegative low-density lipoprotein (LDL) (LDL(-)), a modified LDL that is present in blood and exerts atherogenic effects on endothelial cells and monocytes. This study aimed to determine the action of LDL(-) on monocytes differentiated into macrophages. LDL(-) and in vitro-modified LDLs (oxidized, aggregated, and acetylated) were added to macrophages derived from THP1 monocytes over-expressing CD14 (THP1-CD14). Then, cytokine release, cell differentiation, lipid accumulation, and gene expression were measured by ELISA, flow cytometry, thin-layer chromatography, and real-time PCR, respectively. LDL(-) induced more cytokine release in THP1-CD14 macrophages than other modified LDLs. LDL(-) also promoted morphological changes ascribed to differentiated macrophages. The addition of high-density lipoprotein (HDL) and anti-TLR4 counteracted these effects. LDL(-) was highly internalized by macrophages, and it was the major inductor of intracellular lipid accumulation in triglyceride-enriched lipid droplets. In contrast to inflammation, the addition of anti-TLR4 had no effect on lipid accumulation, thus suggesting an uptake pathway alternative to TLR4. In this regard, LDL(-) upregulated the expression of the scavenger receptors CD36 and LOX-1, as well as several genes involved in triglyceride (TG) accumulation. The importance and novelty of the current study is that LDL(-), a physiologically modified LDL, exerted atherogenic effects in macrophages by promoting differentiation, inflammation, and triglyceride-enriched lipid droplets formation in THP1-CD14 macrophages, probably through different receptors

Sex-Dependent End-of-Life Mental and Vascular Scenarios for Compensatory Mechanisms in Mice with Normal and AD-Neurodegenerative Aging

Life expectancy decreases with aging, with cardiovascular, mental health, and neurodegenerative disorders strongly contributing to the total disability-adjusted life years. Interestingly, the morbidity/mortality paradox points to females having a worse healthy life expectancy. Since bidirectional interactions between cardiovascular and Alzheimer's diseases (AD) have been reported, the study of this emerging field is promising. In the present work, we further explored the cardiovascular-brain interactions in mice survivors of two cohorts of non-transgenic and 3xTg-AD mice, including both sexes, to investigate the frailty/survival through their life span. Survival, monitored from birth, showed exceptionally worse mortality rates in females than males, independently of the genotype. This mortality selection provided a "survivors" cohort that could unveil brain-cardiovascular interaction mechanisms relevant for normal and neurodegenerative aging processes restricted to long-lived animals. The results show sex-dependent distinct physical (worse in 3xTg-AD males), neuropsychiatric-like and cognitive phenotypes (worse in 3xTg-AD females), and hypothalamic-pituitary-adrenal (HPA) axis activation (higher in females), with higher cerebral blood flow and improved cardiovascular phenotype in 3xTg-AD female mice survivors. The present study provides an experimental scenario to study the suggested potential compensatory hemodynamic mechanisms in end-of-life dementia, which is sex-dependent and can be a target for pharmacological and non-pharmacological interventions

Caracterización de la proliferación celular en las zonas subventriculares y de la evolución del infarto mediante espectroscopía de resonancia magnética en un modelo preclínico de isquemia cerebral

El ictus es una patología con una elevada incidencia en la población y los pacientes afectados sufren secuelas que les causan algún tipo de discapacidad. A nivel terapéutico, la posibilidad de incidir sobre la evolución de la enfermedad es a día de hoy aún escasa. Se ha descrito que la neurogénesis, la generación de nuevas neuronas con capacidad para migrar a la zona afectada, aumenta en las zonas subventriculares (ZSVs) y el hipocampo tras un infarto cerebral, en modelos animales y en humanos. Ante este panorama, los estudios dedicados a la comprensión de los mecanismos fisiopatológicos de la neurorreparación, así como el diseño de terapias que incidan sobre dichos mecanismos pueden ser de interés para la mejora de la calidad de vida de los pacientes tras sufrir un ictus. Actualmente se están desarrollando diversas terapias dedicadas a estimular la neurogénesis endógena, la mayoría a nivel preclínico. Una de las necesidades actuales para poder realizar ensayos clínicos sobre neurogénesis en el futuro, sería disponer de biomarcadores que permitieran detectar y monitorizar la neurogénesis de forma no invasiva. Ello nos permitiría conocer las variables asociadas a una mejor o peor capacidad de reparación, observar cómo varía esta capacidad a lo largo del tiempo, cómo varía con la administración de diversos tratamientos y cómo se relaciona esta actividad con el resultado funcional a corto y largo plazo tras un infarto cerebral. La espectroscopía de resonancia magnética (ERM) es una técnica que permite la evaluación bioquímica, in vivo, del tejido de interés de forma no invasiva, incluso permite la obtención de imágenes moleculares de las distribución de un metabolito en el cerebro. No obstante, antes de poder trasladar estos conocimientos al campo clínico, es necesario validar dichos biomarcadores espectroscópicos en modelos animales que permitan correlacionarlos con técnicas inmunohistoquímicas o de microscopía (las actuales técnicas de referencia para la determinación de la neurogénesis). En esta tesis se han desarrollado protocolos de ERM para la caracterización del patrón espectral tanto para las ZSVs como para la zona de infarto en un modelo oclusión transitoria de la arteria cerebral media en rata. Además, para una caracterización metabolómica más precisa, se aplicaron otras técnicas de resonancia que podían permitir una mejor resolución espectral, como el análisis de biopsias de tejido mediante resonancia magnética de alto campo High Resolution Magic Angle Spinning (HRMAS) previo sacrificio de los animales por irradiación con microondas focalizadas (FMW) para detener el metabolismo post mortem. También se utilizaron herramientas informáticas de reconocimiento de patrones para la determinación de los metabolitos con características diferenciales de las ZSVs y la evolución del infarto en base al análisis simultáneo de varios metabolitos. Asimismo, se realizaron pruebas con el fármaco Citicolina para verificar si la técnica permitía detectar cambios en el patrón espectroscópico en las regiones de interés debido a la administración del fármaco. El uso de estas técnicas ha permitido determinar posibles biomarcadores relacionados con apoptosis en la zona de infarto y proliferación en la ZSVs así como la valoración de los efectos de la Citicolina en el patrón metabólico correlacionando los hallazgos espectroscópicos con análisis inmunohistoquímico Los resultados obtenidos ponen de relieve el potencial de la ERM en el análisis bioquímico para determinar biomarcadores con potencial traslacional a la práctica clínica aplicables a futuros estudios en el campo del ictus.Stroke is a disease with a high incidence in the population and the affected patients have sequelae that cause disabilities. From a therapeutic point of view, the possibility of treating the evolution of the disease is nowadays still limited. Neurogenesis, the generation of new neurons with the capacity of migrating to the injured region, has been described to increase in the subventricular zones (SVZs) and hippocampus post-stroke, in animal models and also in humans. That is why studies dedicated to the understanding of the neurorepair physiopathology and the design of new therapies that could influence their mechanisms could be of interest for improving the quality of life of the patients after suffering a stroke. New therapies to promote endogenous neurogenesis enhancement are currently being developed, most of them in preclinical studies. One of the needs to develop neurogenesis clinical trials in the future is to determine biomarkers that would detect and monitor neurogenesis noninvasively. This could be useful to identify associated variables with a better or worse repair capacity, how this capacity changes over time and with pharmacological treatment and how this activity correlates with functional outcome in early and late stages post-stroke. Magnetic resonance spectroscopy (MRS) is a noninvasive technique that allows metabolic profile analysis of the tissue of interest in vivo. Moreover, it allows to perform metabolic imaging of the metabolite distribution through the brain. However, before using spectroscopic biomarkers in the clinical field they must be validated in animal models so that they can be correlated with immunohistochemistry or microscopy (the actual gold standard techniques to determine neurogenesis). In this work MRS protocols were developed to characterize the spectroscopic pattern for the SVZs and also for the stroke zone in a transient middle cerebral artery occlusion rat model. Also, for a more accurate metabolomic characterization with better spectral resolution, other magnetic resonance techniques were applied like biopsy analysis with High Resolution Magic Angle Spinning (HRMAS) using prior focused microwave irradiation (FMW) animal sacrifice in order to arrest post-mortem metabolism. In addition, pattern recognition software was applied to determine feature differences between SVZs or stroke evolution using simultaneous analysis of several metabolites. MRS experiments were performed using a therapeutic agent (Citicoline) to determine if changes in the spectroscopic pattern due to drug administration could be detected. The combination of these techniques has allowed to determine potential biomarkers associated to apoptosis in the stroke zone and proliferation in the SVZs and also to determine Citicoline effects in the metabolic pattern correlating the spectroscopic findings with immunohistochemistry analysis. The results highlight that MRS is a useful technique to perform biochemical analysis to determine biomarkers with translational potential to the clinical practice that could be used for future studies in stroke