ESTUDIO DE LOS MECANISMOS CELULARES EN LA ENDOCARDITIS INFECCIOSA CAUSADA POR STAPHYLOCOCCUS AUREUS: DIFERENCIAS DE GENERO Y NUEVAS DIANAS TERAPÉUTICAS.

PI21/00280

Nombre agencia financiadora Instituto de Salud Carlos III
Acrónimo agencia financiadora ISCIII
Programa Programa Estatal de Generación de Conocimiento y Fortalecimiento del Sistema Español de I+D+I
Subprograma Subprograma Estatal de Generación de Conocimiento
Convocatoria Proyectos de investigación en salud
Año convocatoria 2021
Unidad de gestión Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020 (ISCIII)
Centro beneficiario FUNDACION INSTITUTO DE INVESTIGACION SANITARIA DE NAVARRA
Centro realización INSTITUTO DE INVESTIGACION SANITARIA DE NAVARRA (IdISNA)
Identificador persistente https://doi.org/10.13039/501100004587

Publicaciones

Resultados totales (Incluyendo duplicados): 7
Encontrada(s) 1 página(s)

Chemerin is a new sex-specific target in aortic stenosis concomitant with diabetes regulated by the aldosterone/mineralocorticoid receptor axis

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Goñi Olóriz, Miriam
  • Garaikoetxea Zubillaga, Mattie
  • San Ildefonso-García, Susana
  • Fernández Celis, Amaya
  • Castillo, Paula
  • Navarro, Adela
  • Álvarez, Virginia
  • Sádaba Sagredo, Rafael
  • Jover, Eva
  • Martín Núñez, Ernesto
  • López Andrés, Natalia
Diabetes mellitus (DM) increases the risk of aortic stenosis (AS) and worsens its pathophysiology in a sex-specific manner. Aldosterone/mineralocorticoid receptor (Aldo/MR) pathway participates in the early stages of AS and in other diabetic-related cardiovascular complications. We aim to identify new sex-specific Aldo/MR targets in AS complicated with DM. We performed discovery studies using Olink Proteomics technology in 87 AS patient-derived aortic valves (AVs) (N ¼ 28 and N ¼ 19 nondiabetic and diabetic men; N ¼ 32 and N ¼ 8 nondiabetic and diabetic women, respectively) and human cytokine array (N ¼ 24 AVs/sex/condition). Both approaches revealed chemerin as a target differentially upregulated in AVs from male diabetic patients, further validated in a cohort of stenotic AVs (N ¼ 283, 27.6% DM, 59.4% men). Valvular chemerin levels are directly correlated with valve interstitial cell (VIC) activation, MR, inflammation, angiogenesis, and calcification markers exclusively in diabetic men. In vitro, Aldo (10-8 M) treatment exclusively increased chemerin levels in valve interstitial cells (VICs) from male patients with DM. Aldo also upregulated inflammatory, angiogenic, and osteogenic markers in DM and non-DM donors¿ VICs, which were prevented by MR antagonism. Increased glucose levels in cell media upregulated chemerin in VICs from male diabetic patients. Overall, RARRES2-knockdown in male diabetic VICs resulted in the downregulation of inflammatory, angiogenic, and osteogenic markers and blocked Aldo-induced responses in high glucose conditions. These data suggest the Aldo/MR pathway selectively increases chemerin in VICs from diabetic men, promoting inflammation, angiogenesis, and calcification associated with AS progression., This work was supported by the Instituto de Salud Carlos III Grant PI21/00280 and Departamento de Salud del Gobierno de Navarra Grant GNa01/21. M.G.-O. is supported by a Universidad Pública de Navarra, S.S.I.-G. by a 'Ayudas para la contratación de doctorandos y doctorandas industriales, doctorados industriales 2024' (0011-1408-2024-000006), and M.G.Z. by a Miguel Servet Foundation PhD studentships. E.J. is supported by Roche 'Stop Fuga de cerebros' 2023 and E.M.-N. by Margarita Salas postdoctoral fellowships (ULL-MS-P14, granted by Universidad de La Laguna and Ministerio de Universidades, Gobierno de España).




Sex-specific role of galectin-3 in aortic stenosis

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Matilla Cuenca, Lara
  • Martín Núñez, Ernesto
  • Garaikoetxea Zubillaga, Mattie
  • Navarro, Adela
  • Tamayo Rodríguez, Ibai
  • Fernández Celis, Amaya
  • Gaínza Calleja, Alicia
  • Fernández Irigoyen, Joaquín
  • Santamaría Martínez, Enrique
  • Muntendam, Pieter
  • Álvarez, Virginia
  • Sádaba Sagredo, Rafael
  • Jover, Eva
  • López Andrés, Natalia
Background: Aortic stenosis (AS) is characterized by infammation, fbrosis, osteogenesis and angiogenesis. Men and women develop these mechanisms diferently. Galectin-3 (Gal-3) is a pro-infammatory and pro-osteogenic lectin in AS. In this work, we aim to analyse a potential sex-diferential role of Gal-3 in AS. Methods: 226 patients (61.50% men) with severe AS undergoing surgical aortic valve (AV) replacement were recruited. In AVs, Gal-3 expression and its relationship with infammatory, osteogenic and angiogenic markers was assessed. Valve interstitial cells (VICs) were primary cultured to perform in vitro experiments. Results: Proteomic analysis revealed that intracellular Gal-3 was over-expressed in VICs of male AS patients. Gal-3 secretion was also higher in men’s VICs as compared to women’s. In human AVs, Gal-3 protein levels were signifcantly higher in men, with stronger immunostaining in VICs with myofbroblastic phenotype and valve endothelial cells. Gal-3 levels in AVs were positively correlated with infammatory markers in both sexes. Gal-3 expression was also posi tively correlated with osteogenic markers mainly in men AVs, and with angiogenic molecules only in this sex. In vitro, Gal-3 treatment induced expression of infammatory, osteogenic and angiogenic markers in male’s VICs, while it only upregulated infammatory and osteogenic molecules in women-derived cells. Gal-3 blockade with pharma cological inhibitors (modifed citrus pectin and G3P-01) prevented the upregulation of infammatory, osteogenic and angiogenic molecules. Conclusions: Gal-3 plays a sex-diferential role in the setting of AS, and it could be a new sex-specifc therapeutic target controlling pathological features of AS in VICs., Open Access funding provided by Universidad Pública de Navarra. This work was supported by a Miguel Servet contract (CP13/00221) and by Fondo de Investigaciones Sanitarias (PI18/01875; PI21/00280) from the Instituto de Salud Carlos III—FEDER, and by Departamento de Salud del Gobierno de Navarra (GºNa 01/21). LM was supported by a PFIS Ph.D. studentship (FI19/00302), EM-N was supported by a Margarita Salas postdoctoral fellowship (ULL-MS-P14, granted by Universidad de La Laguna and Ministerio de Universidades, Gobierno de España—Orden UNI/551/2021 de 26 de Mayo and Fondos Next Generation EU), MG was supported by a Miguel Servet Foundation Ph.D. studentship and EJ was supported by a Sara Borrell postdoctoral fellowship (CD19/00251).




Targeting fatty acid-binding protein 4 improves pathologic features of aortic stenosis

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Garaikoetxea Zubillaga, Mattie
  • Martín Núñez, Ernesto
  • Navarro, Adela
  • Matilla Cuenca, Lara
  • Fernández Celis, Amaya
  • Arrieta Paniagua, Vanessa
  • García Peña, Amaia
  • Gaínza Calleja, Alicia
  • Álvarez, Virginia
  • Sádaba Sagredo, Rafael
  • Jover, Eva
  • López Andrés, Natalia
Aortic stenosis (AS) is a fibrocalcific disease of the aortic valves (AVs). Sex-differences in AS pathophysiology have recently been described. High levels of fatty acid-binding protein 4 (FAPB4) in atherosclerotic plaques have been associated with increased local inflammation, endothelial dysfunction, and plaque vulnerability. FABP4 pharmacological blockade has been shown to be effective for the treatment of atherosclerosis by modulating metabolic and inflammatory pathways. We aimed to analyze the sex-specific expression of FABP4 in AS and its potential role as a therapeutic target. A total of 226 patients (61.5% men) with severe AS undergoing surgical AV replacement were recruited. The FABP4 levels were increased in the AVs of AS patients compared to the control subjects, showing greater expression in the fibrocalcific regions. Male AVs exhibited higher levels of FABP4 compared to females, correlating with markers of inflammation (IL-6, Rantes), apoptosis (Bax, caspase-3, Bcl-2), and calcification (IL-8, BMP-2 and BMP-4). VICs derived from AS patients showed the basal expression of FABP4 in vitro. Osteogenic media induced upregulation of intracellular and secreted FABP4 levels in male VICs after 7 days, along with increased levels of inflammatory, pro-apoptotic, and osteogenic markers. Treatment with BMS309403, a specific inhibitor of FABP4, prevented from all of these changes. Thus, we propose FABP4 as a new sex-specific pharmacological therapeutic target in AS., This research was funded by Miguel Servet contract CP13/00221 from the Instituto de Salud Carlos III-FEDER, Fondo de Investigaciones Sanitarias [PI18/01875; PI21/00280]. M.G. is supported by a Miguel Servet Foundation PhD studentship, E.M.-N. is supported by a Margarita Salas postdoctoral fellowship (ULL-MS-P14), L.M. is supported by a PFIS (FI19/00302) PhD studentship, E.J. (CD19/00251) is supported by a Sara Borrell postdoctoral fellowship.




Characterization of the sex-specific pattern of angiogenesis and lymphangiogenesis in aortic stenosis

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Matilla Cuenca, Lara
  • Martín Núñez, Ernesto
  • Garaikoetxea Zubillaga, Mattie
  • Navarro, Adela
  • Vico, Julieta Anabela
  • Arrieta Paniagua, Vanessa
  • García Peña, Amaia
  • Fernández Celis, Amaya
  • Gaínza Calleja, Alicia
  • Álvarez, Virginia
  • Sádaba Sagredo, Rafael
  • López Andrés, Natalia
  • Jover, Eva
Objective: We aim to analyze sex-related differences in angiogenesis and lymphangiogenesis in aortic valves (AVs) and valve interstitial cells (VICs) from aortic stenosis (AS) patients. Approach and Results: Totally 230 patients (59% men) with severe AS undergoing surgical valve replacement were recruited. The density of total neovessels was higher in AVs from men as compared to women. Both small and medium neovessels were more abundant in men's AVs. Accordingly, male AVs exhibited higher CD31 and VE-cadherin expressions. The levels of the pro-angiogenic markers, such as vascular endothelial growth factor (VEGF)-A, VEGF receptor (VEGFR)1, VEGFR2, insulin-like growth factor-binding protein-2 (IGFBP-2), interleukin (IL)-8, chemerin, and fibroblast growth factor (FGF)-7, were increased in AVs from men. Transforming growth factor-¿ expression was higher in male AVs. The expression of antiangiogenic molecules thrombospondin (Tsp)-1, endostatin, and CD36 was upregulated in male AVs, although the levels of Tsp-2, IL-4, IL-12p70, and chondromodulin-1 were similar between both sexes. The number of lymphatic vessels and the expression of the lymphangiogenic markers Lyve-1 and D2-40 was higher in men's AV as well as VEGF-C, VEGF-D, and VEGFR3. Multivariate analyses adjusted for confounders further validated the sex-dependent expression of these targets. VICs isolated from men's AVs secreted higher amounts of the pro-angiogenic factors, VEGF-A, VEGFR1, IGFBP-2, and FGF-7, as well as the pro-lymphangiogenic factors, VEGF-C, VEGF-D, and VEGFR3, than women without changes in antiangiogenic markers. Conclusion: Our data show that aberrant angiogenic and lymphangiogenic cues are over-represented in male AVs. Importantly, the VIC is a relevant source of multiple morphogens involved in angiogenesis and lymphangiogenesis likely endowing the AV of men with the predominant calcific AS phenotypes., This work was supported by a Miguel Servet contract (CP13/00221) and by Fondo de Investigaciones Sanitarias (PI18/01875; PI21/00280) from the Instituto de Salud Carlos III - FEDER. LM was supported by a PFIS Ph.D. studentship (FI19/00302). EJ was supported by a Sara Borrell postdoctoral fellowship (CD19/00251). EM-N was supported by a Margarita Salas postdoctoral fellowship (ULL-MS-P14). MG was supported by a Miguel Servet Foundation Ph.D. studentship.




Influence of diabetes mellitus on the pathological profile of aortic stenosis: a sex-based approach

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Martín Núñez, Ernesto
  • Goñi Olóriz, Miriam
  • Matilla Cuenca, Lara
  • Garaikoetxea Zubillaga, Mattie
  • Mourino-Álvarez, Laura
  • Navarro, Adela
  • Fernández Celis, Amaya
  • Tamayo Rodríguez, Ibai
  • Gaínza Calleja, Alicia
  • Álvarez, Virginia
  • Sádaba Sagredo, Rafael
  • Barderas, María G.
  • Jover, Eva
  • López Andrés, Natalia
Background: Diabetes mellitus (DM) accelerates the progression of aortic stenosis (AS), but how their underlying molecular mechanisms interact is not clear. Moreover, whether DM contributes to clinically relevant sex-differences in AS is unknown. In this work we aim to characterize the sex-specific profile of major pathological mechanisms fundamental to aortic valve (AV) degeneration in AS patients with or without concomitant DM. Methods: 283 patients with severe AS undergoing surgical valve replacement (27.6% DM, 59.4% men) were recruited. Expression of pathological markers related to AS were thoroughly assessed in AVs and valve interstitial cells (VICs) according to sex and presence of DM. Complementary in vitro experiments in VICs in the presence of high-glucose levels (25 mM) for 24, 48 and 72 h were performed. Results: Oxidative stress and metabolic dysfunction markers were increased in AVs from diabetic AS patients compared to non-diabetic patients in both sexes. However, disbalanced oxidative stress and enhanced inflammation were more predominant in AVs from male AS diabetic patients. Osteogenic markers were exclusively increased in the AVs of diabetic women. Basal characterization of VICs confirmed that oxidative stress, inflammation, calcification, and metabolic alteration profiles were increased in diabetic VICs with sex-specific differences. VICs cultured in hyperglycemic-like conditions triggered inflammatory responses in men, whereas in women rapid and higher production of pro-osteogenic molecules. Conclusions: DM produces sex-specific pathological phenotypes in AV of AS patients. Importantly, women with diabetes are more prone to develop AV calcification. DM should be considered as a risk factor in AS especially in women., This work was supported by a Miguel Servet contract (CP13/00221) and Fondo de Investigaciones Sanitarias (PI21/00280) from the Instituto de Salud Carlos III – FEDER, as well as by Gobierno de Navarra (GºNa 01/21). EMN was supported by a Margarita Salas postdoctoral fellowship (ULL-MS-P14, granted by Universidad de la Laguna and Ministerio de Universidades - Orden UNI/551/2021 de 26 de mayo and Fondos Next Generation EU), MG was supported by a Miguel Servet Foundation Ph.D. studentship, LM was supported by a PFIS Ph.D. studentship (FI19/00302) and EJ was supported by a Sara Borrell postdoctoral fellowship (CD19/00251). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Open Access funding provided by Universidad Pública de Navarra.




Sex-dependent expression of neutrophil gelatinase-associated lipocalin in aortic stenosis

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Jover, Eva
  • Matilla Cuenca, Lara
  • Martín Núñez, Ernesto
  • Garaikoetxea Zubillaga, Mattie
  • Navarro, Adela
  • Fernández Celis, Amaya
  • Gaínza Calleja, Alicia
  • Arrieta Paniagua, Vanessa
  • García Peña, Amaia
  • Álvarez, Virginia
  • Sádaba Sagredo, Rafael
  • Jaisser, Frederic
  • López Andrés, Natalia
Background: Accumulating evidence suggest the existence of sex-related differences in the pathogenesis of aortic stenosis (AS) with inflammation, oxidative stress, fibrosis and calcification being over-represented in men. Neutrophil gelatinase-associated lipocalin (NGAL) is expressed in a myriad of tissues and cell types, and it is associated with acute and chronic pathological processes comprising inflammation, fibrosis or calcification. Sex-dependent signatures have been evidenced for NGAL which expression has been associated predominantly in males to metabolic and cardiovascular disorders. We aimed to analyse sex-related differences of NGAL in AS and its role in the inflammatory and fibrocalcific progression of AS. Methods and results: 220 (60.45% men) patients with severe AS elective for surgical aortic valve (AV) replacement were recruited. Immunohistochemistry revealed higher expression of NGAL in calcific areas of AVs and that was validated by qPCR in in 65 (60% men) donors. Valve interstitial cells (VICs) were a source of NGAL in these samples. Proteome profiler analyses evidenced higher expression of NGAL in men compared to women, and that was further validated by ELISA. NGAL expression in the AV was correlated with inflammation, oxidative stress, and osteogenic markers, as well as calcium score. The expression of NGAL, both intracellular and secreted (sNGAL), was significantly deregulated only in calcifying male-derived VICs. Depletion of intracellular NGAL in calcifying male-derived VICs was associated with pro-inflammatory profiles, dysbalanced matrix remodelling and pro-osteogenic profiles. Conversely, exogenous NGAL mediated inflammatory and dysbalanced matrix remodelling in calcifying VICs, and all that was prevented by the pharmacological blockade of NGAL. Conclusions: Owing to the over-expression of NGAL, the AV from men may be endowed with higher expression of inflammatory, oxidative stress, matrix remodelling and osteogenic markers supporting the progression of calcific AS phenotypes. The expression of NGAL in the VIC emerges as a potential therapeutic checkpoint, with its effects being potentially reverted by the pharmacological blockade of extracellular NGAL., This research was funded by Miguel Servet contract CP13/00221 from Instituto de Salud Carlos III-FEDER, Fondo de Investigaciones Sanitarias [PI18/01875; PI21/00280]. M.G. is supported by a Miguel Servet Foundation PhD studentship, E.M.-N is supported by a Margarita Salas postdoctoral fellowship (ULL-MS-P14), L.M. is supported by a PFIS (FI19/00302) PhD studentship, E.J. (CD19/00251) is supported by a Sara Borrell postdoctoral fellowship.




Sex differences in aortic valve inflammation and remodeling in chronic severe aortic regurgitation

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Tiraplegui, Carolina
  • Garaikoetxea Zubillaga, Mattie
  • Sádaba, Alba
  • San Ildefonso-García, Susana
  • Goñi Olóriz, Miriam
  • Fernández Celis, Amaya
  • Martín Núñez, Ernesto
  • Álvarez, Virginia
  • Sádaba Sagredo, Rafael
  • Anand, Vidhu
  • Jover, Eva
  • Navarro, Adela
  • López Andrés, Natalia
Aortic regurgitation (AR) is more prevalent in males, although cellular and molecular mechanisms underlying the sex differences in prevalence and pathophysiology are unknown. This study evaluates the impact of sex on aortic valve (AV) inflammation and remodeling and the cellular differences in valvular interstitial cells (VICs) and valvular endothelial cells (VECs) in patients with AR. A total of 144 patients (27.5% female) with severe chronic AR were included. AVs were analyzed by imaging, histological, and molecular biology techniques (ELISA, RT-PCR). VICs and VECs isolated from patients with AR were characterized and further treated with transforming growth factor (TGF)-β. Anatomically, male had smaller index aortic dimensions and greater AV thickness. Proteome profiler analyzes in AVs (n = 40/sex) evidenced higher expression of inflammatory markers in male and that was further validated (interleukins, chemokines). Histological composition showed higher expression of inflammatory mediators and collagen thick fibers in AVs from male. Male VICs and VECs secreted higher levels of inflammatory markers than female cells. Interestingly, male VICs produced higher amounts of collagen type I and lower fibronectin and aggrecan, whereas male VECs secreted lower decorin. TGF-β exclusively enhanced inflammation in male VICs and decorin and aggrecan in female VICs. Compared with male, AVs from female were thinner, less inflamed, and fibrotic. VICs seem to be the key cell type responsible for the sex-differences. Valvular inflammation associated with an active remodeling process could be a key pathophysiological process involved in AR., This research was funded by Fondo de Investigaciones Sanitarias under Grant Nos. PI18/01875; PI21/00280 from Instituto de Salud Carlos III-FEDER and BIOGEN (PC020-021-022) from Ayudas a Agentes del SINAI para la realización de proyectos de I+D colaborativos del Gobierno de Navarra. M.G. is supported by a Miguel Servet Foundation PhD studentship, E.M.-N is supported by a Margarita Salas postdoctoral fellowship ULL-MS-P14 and E.J. (CD19/00251) was supported by a Sara Borrell postdoctoral fellowship and by a Roche 'Stop Fuga de cerebros' postdoctoral fellowship. This study was also funded by the Departamento de Salud del Gobierno de Navarra Grant GºNa01/21.