Role of the Scavenger Receptor CD36 in Accelerated Diabetic Atherosclerosis

Diabetes mellitus entails increased atherosclerotic burden and medial arterial calcification, but the precise mechanisms are not fully elucidated. We aimed to investigate the implication of CD36 in inflammation and calcification processes orchestrated by vascular smooth muscle cells (VSMCs) under hyperglycemic and atherogenic conditions. We examined the expression of CD36, pro-inflammatory cytokines, endoplasmic reticulum (ER) stress markers, and mineralization-regulating enzymes by RT-PCR in human VSMCs, cultured in a medium containing normal (5 mM) or high glucose (22 mM) for 72 h with or without oxidized low-density lipoprotein (oxLDL) (24 h). The uptake of 1,1'-dioctadecyl-3,3,3',3-tetramethylindocarbocyanine perchlorate-fluorescently (DiI) labeled oxLDL was quantified by flow cytometry and fluorimetry and calcification assays were performed in VSMC cultured in osteogenic medium and stained by alizarin red. We observed induction in the expression of CD36, cytokines, calcification markers, and ER stress markers under high glucose that was exacerbated by oxLDL. These results were confirmed in carotid plaques from subjects with diabetes versus non-diabetic subjects. Accordingly, the uptake of DiI-labeled oxLDL was increased after exposure to high glucose. The silencing of CD36 reduced the induction of CD36 and the expression of calcification enzymes and mineralization of VSMC. Our results indicate that CD36 signaling is partially involved in hyperglycemia and oxLDL-induced vascular calcification in diabetes

Oxidative Stress and Inflammatory Markers in Abdominal Aortic Aneurysm.

Abdominal aortic aneurysm (AAA) is increasing due to aging of the population and is a major cause of death among the elderly. Ultrasound screening programs are useful in early diagnosis, but aneurysm size is not always a good predictor of rupture. Our aim was to analyze the value of circulating molecules related to oxidative stress and inflammation as new biomarkers to assist the management of AAA. The markers were quantified by ELISA, and their expression in the aneurysmal wall was studied by real-time PCR and by immunostaining. Correlation analysis of the studied markers with aneurysm diameter and peak wall stress (PWS), obtained by finite element analysis, and multivariate regression analysis to assess potential confounding factors were performed. Our study shows an extensive inflammatory infiltration in the aneurysmal wall, mainly composed by T-cells, macrophages and B-cells and altered levels of reactive oxygen species (ROS), IgM, IgG, CD38, GDF15, S100A4 and CD36 in plasma and in the aneurysmal tissue of AAA patients compared with controls. Circulating levels of IgG, CD38 and GDF15 positively correlated with abdominal aortic diameter, and CD38 was correlated with PWS. Our data show that altered levels of IgG, CD38 and GDF15 have potential diagnostic value in the assessment of AAA

Oxidative stress and inflammatory markers in abdominal aortic aneurysm

Abdominal aortic aneurysm (AAA) is increasing due to aging of the population and is a major cause of death among the elderly. Ultrasound screening programs are useful in early diagnosis, but aneurysm size is not always a good predictor of rupture. Our aim was to analyze the value of circulating molecules related to oxidative stress and inflammation as new biomarkers to assist the management of AAA. The markers were quantified by ELISA, and their expression in the aneurysmal wall was studied by real-time PCR and by immunostaining. Correlation analysis of the studied markers with aneurysm diameter and peak wall stress (PWS), obtained by finite element analysis, and multivariate regression analysis to assess potential confounding factors were performed. Our study shows an extensive inflammatory infiltration in the aneurysmal wall, mainly composed by T-cells, macrophages and B-cells and altered levels of reactive oxygen species (ROS), IgM, IgG, CD38, GDF15, S100A4 and CD36 in plasma and in the aneurysmal tissue of AAA patients compared with controls. Circulating levels of IgG, CD38 and GDF15 positively correlated with abdominal aortic diameter, and CD38 was correlated with PWS. Our data show that altered levels of IgG, CD38 and GDF15 have potential diagnostic value in the assessment of AAA

Antibodies against peripheral nerve antigens in chronic inflammatory demyelinating polyradiculoneuropathy

Altres ajuts: Beca Juan Rodes JR1300014Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is a heterogeneous disease in which diverse autoantibodies have been described but systematic screening has never been performed. Detection of CIDP-specific antibodies may be clinically useful. We developed a screening protocol to uncover novel reactivities in CIDP. Sixty-five CIDP patients and 28 controls were included in our study. Three patients (4.6%) had antibodies against neurofascin 155, four (6.2%) against contactin-1 and one (1.5%) against the contactin-1/contactin-associated protein-1 complex. Eleven (18.6%) patients showed anti-ganglioside antibodies, and one (1.6%) antibodies against peripheral myelin protein 2. No antibodies against myelin protein zero, contactin-2/contactin-associated protein-2 complex, neuronal cell adhesion molecule, gliomedin or the voltage-gated sodium channel were detected. In IgG experiments, three patients (5.3%) showed a weak reactivity against motor neurons; 14 (24.6%) reacted against DRG neurons, four of them strongly (7.0%), and seven (12.3%) reacted against Schwann cells, three of them strongly (5.3%). In IgM experiments, six patients (10.7%) reacted against DRG neurons, while three (5.4%) reacted against Schwann cells. However, results were not statistically significant when compared to controls. Immunoprecipitation experiments identified CD9 and L1CAM as potential antigens, but reactivity could not be confirmed with cell-based assays. In summary, we describe a diverse autoantibody repertoire in CIDP patients, reinforcing the hypothesis of CIDP's pathophysiological heterogeneity

Nuevas estrategias terapéuticas para limitar el desarrollo del aneurisma de aorta abdominal: inhibición del estrés del retículo endoplasmático y mitocondrial

Introducción: el aneurisma de aorta abdominal (AAA) es una enfermedad vascular degenerativa caracterizada por una dilatación de la aorta abdominal del 50% o mayor que el diámetro normal. Presenta una prevalencia del 4-8 % en hombres mayores de 65 años y está asociada a una alta morbilidad y mortalidad. Es una enfermedad con una etiopatogenia muy compleja y se cree causada por una combinación de múltiples factores de riesgo entre los que se incluye el tabaquismo, edad avanzada, hipertensión, aterosclerosis y tener antecedentes familiares. Objetivo: actualmente no existen terapias efectivas para frenar el desarrollo del AAA y su tratamiento se limita a la reparación quirúrgica. En la literatura se ha descrito la contribución de la activación del estrés del retículo endoplasmático (RE) y su relación con la disfunción mitocondrial con la fisiopatología de diversas enfermedades cardiovasculares. Nuestro objetivo es dilucidar la posible implicación del estrés del RE en el desarrollo del AAA a través del estudio de muestras de aortas aneurismáticas procedentes de una cohorte de pacientes y determinar la eficacia de la inhibición del estrés del RE para limitar la degeneración vascular en el AAA. Materiales y métodos: la expresión de marcadores del estrés del RE, del estrés mitocondrial y de fenómenos de autofagia y apoptosis se analizó mediante PCR a tiempo real, western-blot e inmunotinción en muestras humanas de AAA. La cuantificación de oxisteroles en plasma se realizó por cromatrografía líquida acoplada a espectrometría de masas (LC-(APCI)-MS/MS) y los niveles de S100A4 y Gdf15 se cuantificaron mediante kits de ELISA. Las características de la pared vascular se estudiaron mediante técnicas de histología clásica y el infiltrado inflamatorio se estudió mediante inmunohistoquímica en la aorta abdominal en nuestra cohorte pacientes y donantes y en el modelo murino de AAA (deficiente en apolipoproteínaE infundido con angiotensina II) utilizado en nuestro estudio. En este modelo se evaluó el efecto de la administración de inhibidores del estrés del RE, fenilbutirato sódico (PBA) y ácido tauroursodeoxicólico (TUDCA), sobre el diámetro aórtico en los días 0, 3, 7, 14, 21 y 28 de la infusión con Ang II mediante ultrasonografía y sobre la presión arterial (método del manguito en la cola). La expresión de marcadores del estrés del RE, de apoptosis y de inflamación y de las metaloproteinasas (MMPs) 2 y 9 se cuantificó mediante PCR a tiempo real y en el caso de las MMPs se realizaron ensayos de zimografía clásica para determinar su actividad gelatinolítica en la aorta abdominal de ratones. Resultados: en pacientes con AAA y en la aorta abdominal del ratón ApoE-/- infundido con Ang II se confirmó el aumento de la expresión génica y proteica de diferentes marcadores del estrés del RE acompañado de una exacerbación del estrés oxidativo, un descenso en la expresión de indicadores de la biogénesis mitocondrial y un incremento en la apoptosis en la pared vascular. En plasma, el perfil de oxisteroles y los niveles de S100A4 y Gdf15 se encontraron alterados en pacientes respecto a donantes. La administración de PBA y TUDCA redujo la incidencia de AAA disminuyendo la dilatación aórtica y la desestructuración de la pared vascular y redujo significativamente la presión arterial. La sobre-expresión de los marcadores de estrés del RE, de inflamación y de metaloproteinasas en la aorta abdominal de animales disminuyó con el tratamiento con PBA y TUDCA de manera diferencial. Conclusiones: Nuestros resultados evidencian la activación de estrés del RE en el AAA humano y sugieren que su inhibición podría constituir una estrategia terapéutica para limitar del desarrollo de esta enfermedad.Introduction: the abdominal aortic aneurysm (AAA) is a degenerative vascular disease characterized by a dilatation of the abdominal aorta of 50% or greater than the normal diameter. It has a prevalence of 4-8% in men older than 65 years and it is associated with a high morbidity and mortality. It is a very complex disease and is believed to be caused by a combination of multiple risk factors: smoking, advanced age, hypertension, atherosclerosis and a family history. Objective: Currently there are no effective therapies to stop the development of AAA and its treatment is limited to surgical repair. The contribution of stress activation of the endoplasmic reticulum (ER) and its relationship with mitochondrial dysfunction to the pathophysiology of several cardiovascular diseases has been previously described. Our objective is to evaluate the possible implication of ER stress in the development of AAA and to test the efficacy of ER stress inhibition to limit vascular degeneration in AAA disease. Materials and methods: the expression of ER stress markers, mitochondrial stress and apoptosis and autophagy markers was analyzed by real-time PCR, western-blot and immunostaining in human AAA samples. The quantification of oxysterols in plasma was performed by liquid chromatography coupled to mass spectrometry (LC- (APCI) -MS / MS) and the circulating levels of S100A4 and Gdf15 were quantified with ELISA kits. The structure of the vascular wall was studied by classical histology techniques and the inflammatory infiltrate was studied by immunohistochemistry in the abdominal aorta of our patients and donors cohort and in the abdominal aorta of our murine model of AAA (mice deficient in apolipoprotein E infused with angiotensin II). In this model we evaluated the effect of the administration of ER stress inhibitors, sodium phenylbutyrate (PBA) and tauroursodeoxicolic acid (TUDCA), on the aortic diameter on days 0, 3, 7, 14, 21 and 28 by ultrasonography during the time of infusion with Ang II and the effect on blood pressure (tail cuff method). The expression of ER stress markers, apoptosis, inflammatory markers and metalloproteinases (MMPs) 2 and 9 was quantified by real-time PCR and in the case of MMPs, zymography tests were performed to determine its gelatinolytic activity in the protein lysates of abdominal aortas from mice. Results: in aneurismatic wall of patients with AAA and in the abdominal aorta of the ApoE-/- mouse infused with Ang II, an increase of the gene and protein expression of different ER stress markers was confirmed. This was accompanied by an exacerbation of oxidative stress, a decrease in the expression of indicators of mitochondrial biogenesis and by an increase in apoptosis in the vascular wall. In plasma, the profile of oxysterols and the levels of S100A4 and Gdf15 were found to be altered in patients vs. donors. In animals, the administration of PBA and TUDCA reduced the incidence of AAA by decreasing aortic dilatation and the disorganization of the vascular wall and by reducing blood pressure. The expression of ER stress markers, inflammatory markers and matrix metalloproteinases in the abdominal aorta of mice decreased with either the treatment with TUDCA or PBA. Conclusions: our results evidence that ER stress is involved in the pathophysiology of aneurysm and show that inhibition of ER stress is effective to limit abdominal aortic dilatation in AAA disease

SS-31 for use in the treatment and/or prevention of aneurysm

The present invention relates to SS-31 or composition comprising SS-31 for use in the treatment and/or prevention of aneurysm.Peer reviewedFundacio Institut De Recerca Del Hospital De La Santa Creu I Sant Pau, Consejo Superior de Investigaciones Científicas (España)A1 Solicitud de patente con informe sobre el estado de la técnic

SS-31 for use in the treatment and/or prevention of aneurysm

The present invention relates to SS-31 or composition comprising SS-31 for use in the treatment and/or prevention of aneurysmPeer reviewedFundacio Institut De Recerca Del Hospital De La Santa Creu I Sant Pau, Consejo Superior de Investigaciones Científicas (España)A1 Solicitud de patente con informe sobre el estado de la técnic

Nuevas estrategias terapéuticas para limitar el desarrollo del aneurisma de aorta abdominal : inhibición del estrés del retículo endoplasmático y mitocondrial /

Departament responsable de la tesi: Departament de Bioquímica i Biologia Molecular.Introducción: el aneurisma de aorta abdominal (AAA) es una enfermedad vascular degenerativa caracterizada por una dilatación de la aorta abdominal del 50% o mayor que el diámetro normal. Presenta una prevalencia del 4-8 % en hombres mayores de 65 años y está asociada a una alta morbilidad y mortalidad. Es una enfermedad con una etiopatogenia muy compleja y se cree causada por una combinación de múltiples factores de riesgo entre los que se incluye el tabaquismo, edad avanzada, hipertensión, aterosclerosis y tener antecedentes familiares.Objetivo: actualmente no existen terapias efectivas para frenar el desarrollo del AAA y su tratamiento se limita a la reparación quirúrgica. En la literatura se ha descrito la contribución de la activación del estrés del retículo endoplasmático (RE) y su relación con la disfunción mitocondrial con la fisiopatología de diversas enfermedades cardiovasculares. Nuestro objetivo es dilucidar la posible implicación del estrés del RE en el desarrollo del AAA a través del estudio de muestras de aortas aneurismáticas procedentes de una cohorte de pacientes y determinar la eficacia de la inhibición del estrés del RE para limitar la degeneración vascular en el AAA.Materiales y métodos: la expresión de marcadores del estrés del RE, del estrés mitocondrial y de fenómenos de autofagia y apoptosis se analizó mediante PCR a tiempo real, western-blot e inmunotinción en muestras humanas de AAA. La cuantificación de oxisteroles en plasma se realizó por cromatrografía líquida acoplada a espectrometría de masas (LC-(APCI)-MS/MS) y los niveles de S100A4 y Gdf15 se cuantificaron mediante kits de ELISA. Las características de la pared vascular se estudiaron mediante técnicas de histología clásica y el infiltrado inflamatorio se estudió mediante inmunohistoquímica en la aorta abdominal en nuestra cohorte pacientes y donantes y en el modelo murino de AAA (deficiente en apolipoproteínaE infundido con angiotensina II) utilizado en nuestro estudio. En este modelo se evaluó el efecto de la administración de inhibidores del estrés del RE, fenilbutirato sódico (PBA) y ácido tauroursodeoxicólico (TUDCA), sobre el diámetro aórtico en los días 0, 3, 7, 14, 21 y 28 de la infusión con Ang II mediante ultrasonografía y sobre la presión arterial (método del manguito en la cola). La expresión de marcadores del estrés del RE, de apoptosis y de inflamación y de las metaloproteinasas (MMPs) 2 y 9 se cuantificó mediante PCR a tiempo real y en el caso de las MMPs se realizaron ensayos de zimografía clásica para determinar su actividad gelatinolítica en la aorta abdominal de ratones.Resultados: en pacientes con AAA y en la aorta abdominal del ratón ApoE-/- infundido con Ang II se confirmó el aumento de la expresión génica y proteica de diferentes marcadores del estrés del RE acompañado de una exacerbación del estrés oxidativo, un descenso en la expresión de indicadores de la biogénesis mitocondrial y un incremento en la apoptosis en la pared vascular. En plasma, el perfil de oxisteroles y los niveles de S100A4 y Gdf15 se encontraron alterados en pacientes respecto a donantes. La administración de PBA y TUDCA redujo la incidencia de AAA disminuyendo la dilatación aórtica y la desestructuración de la pared vascular y redujo significativamente la presión arterial. La sobre-expresión de los marcadores de estrés del RE, de inflamación y de metaloproteinasas en la aorta abdominal de animales disminuyó con el tratamiento con PBA y TUDCA de manera diferencial.Conclusiones: Nuestros resultados evidencian la activación de estrés del RE en el AAA humano y sugieren que su inhibición podría constituir una estrategia terapéutica para limitar del desarrollo de esta enfermedad.Introduction: the abdominal aortic aneurysm (AAA) is a degenerative vascular disease characterized by a dilatation of the abdominal aorta of 50% or greater than the normal diameter. It has a prevalence of 4-8% in men older than 65 years and it is associated with a high morbidity and mortality. It is a very complex disease and is believed to be caused by a combination of multiple risk factors: smoking, advanced age, hypertension, atherosclerosis and a family history. Objective: Currently there are no effective therapies to stop the development of AAA and its treatment is limited to surgical repair. The contribution of stress activation of the endoplasmic reticulum (ER) and its relationship with mitochondrial dysfunction to the pathophysiology of several cardiovascular diseases has been previously described. Our objective is to evaluate the possible implication of ER stress in the development of AAA and to test the efficacy of ER stress inhibition to limit vascular degeneration in AAA disease. Materials and methods: the expression of ER stress markers, mitochondrial stress and apoptosis and autophagy markers was analyzed by real-time PCR, western-blot and immunostaining in human AAA samples. The quantification of oxysterols in plasma was performed by liquid chromatography coupled to mass spectrometry (LC- (APCI) -MS / MS) and the circulating levels of S100A4 and Gdf15 were quantified with ELISA kits. The structure of the vascular wall was studied by classical histology techniques and the inflammatory infiltrate was studied by immunohistochemistry in the abdominal aorta of our patients and donors cohort and in the abdominal aorta of our murine model of AAA (mice deficient in apolipoprotein E infused with angiotensin II). In this model we evaluated the effect of the administration of ER stress inhibitors, sodium phenylbutyrate (PBA) and tauroursodeoxicolic acid (TUDCA), on the aortic diameter on days 0, 3, 7, 14, 21 and 28 by ultrasonography during the time of infusion with Ang II and the effect on blood pressure (tail cuff method). The expression of ER stress markers, apoptosis, inflammatory markers and metalloproteinases (MMPs) 2 and 9 was quantified by real-time PCR and in the case of MMPs, zymography tests were performed to determine its gelatinolytic activity in the protein lysates of abdominal aortas from mice. Results: in aneurismatic wall of patients with AAA and in the abdominal aorta of the ApoE-/- mouse infused with Ang II, an increase of the gene and protein expression of different ER stress markers was confirmed. This was accompanied by an exacerbation of oxidative stress, a decrease in the expression of indicators of mitochondrial biogenesis and by an increase in apoptosis in the vascular wall. In plasma, the profile of oxysterols and the levels of S100A4 and Gdf15 were found to be altered in patients vs. donors. In animals, the administration of PBA and TUDCA reduced the incidence of AAA by decreasing aortic dilatation and the disorganization of the vascular wall and by reducing blood pressure. The expression of ER stress markers, inflammatory markers and matrix metalloproteinases in the abdominal aorta of mice decreased with either the treatment with TUDCA or PBA. Conclusions: our results evidence that ER stress is involved in the pathophysiology of aneurysm and show that inhibition of ER stress is effective to limit abdominal aortic dilatation in AAA disease

Enhanced endoplasmic reticulum and mitochondrial stress in abdominal aortic aneurysm

Abdominal aortic aneurysm (AAA) is a degenerative vascular disease with a complex aetiology that remains to be fully elucidated. Clinical management of AAA is limited to surgical repair, while an effective pharmacotherapy is still awaited. Endoplasmic reticulum (ER) stress and mitochondrial dysfunction have been involved in the pathogenesis of cardiovascular diseases (CVDs), although their contribution to AAA development is uncertain. Therefore, we aimed to determine their implication in AAA and investigated the profile of oxysterols in plasma, specifically 7-ketocholesterol (7-KC), as an ER stress inducer.In the present study, we determined aortic ER stress activation in a large cohort of AAA patients compared with healthy donors. Higher gene expression of activating transcription factor (ATF) 6 (ATF6), IRE-1, X-binding protein 1 (XBP-1), C/EBP-homologous protein (CHOP), CRELD2 and suppressor/enhancer of Lin-12-like (SEL1L) and greater protein levels of active ATF6, active XBP1 and of the pro-apoptotic protein CHOP were detected in human aneurysmatic samples. This was accompanied by an exacerbated apoptosis, higher reactive oxygen species (ROS) production and by a reduction in mitochondrial biogenesis in the vascular wall of AAA. The quantification of oxysterols, performed by liquid chromatography-(atmospheric pressure chemical ionization (APCI))-mass spectrometry, showed that levels of 7-KC were significantly higher while those of 7α-hydroxycholesterol (HC), 24-HC and 27-HC were lower in AAA patients compared with healthy donors. Interestingly, the levels of 7-KC correlate with the expression of ER stress markers.Our results evidence an induction of ER stress in the vascular wall of AAA patients associated with an increase in circulating 7-KC levels and a reduction in mitochondrial biogenesis suggesting their implication in the pathophysiology of this disease.This work was supported by the Spanish Ministerio de Economía y Competitividad (MINECO)-Instituto de Salud Carlos III (ISCIII) [grant numbers CP15/00126, PI17/08137 (to M.G.), PI18/0919 (to C.R.) and RTI2018-094727-B-100 (to J.M.G.)]; the CIBERCV [grant number CB16/11/00257]; the ISCIII, Miguel Servet I program (grant number CP15/00126 (to M.N.M. and M.G.)]; and by Agencia de Gestio d’Ajuts Universitaris i de Recerca (AGAUR; program of support to Research Groups. Ref. 2017-SGR-00333). The study was co-founded by Fondo Europeo de Desarrollo Regional (FEDER)-The way to build Europ

Oxidative Stress and Inflammatory Markers in Abdominal Aortic Aneurysm

Abdominal aortic aneurysm (AAA) is increasing due to aging of the population and is a major cause of death among the elderly. Ultrasound screening programs are useful in early diagnosis, but aneurysm size is not always a good predictor of rupture. Our aim was to analyze the value of circulating molecules related to oxidative stress and inflammation as new biomarkers to assist the management of AAA. The markers were quantified by ELISA, and their expression in the aneurysmal wall was studied by real-time PCR and by immunostaining. Correlation analysis of the studied markers with aneurysm diameter and peak wall stress (PWS), obtained by finite element analysis, and multivariate regression analysis to assess potential confounding factors were performed. Our study shows an extensive inflammatory infiltration in the aneurysmal wall, mainly composed by T-cells, macrophages and B-cells and altered levels of reactive oxygen species (ROS), IgM, IgG, CD38, GDF15, S100A4 and CD36 in plasma and in the aneurysmal tissue of AAA patients compared with controls. Circulating levels of IgG, CD38 and GDF15 positively correlated with abdominal aortic diameter, and CD38 was correlated with PWS. Our data show that altered levels of IgG, CD38 and GDF15 have potential diagnostic value in the assessment of AAA