Publication: Papel de las isomorfas de endoglina en el linaje mieloide
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2014-06-25
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Universidad Complutense de Madrid
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Abstract
Endoglina es una glicoproteína de membrana con un papel crucial en el remodelado vascular y la angiogénesis. Habiéndose descrito su función e implicaciones fisiopatológicas principalmente en el ámbito de la célula endotelial, su papel en el linaje mieloide, donde también se expresa, está todavía por elucidar. En la presente memoria se aborda el estudio de las dos isoformas de endoglina en la línea promonocítica U937, mediante análisis proteómico SILAC. Al analizar las proteínas alteradas debido a la acción de las isoformas de endoglina, se ha identificado a la isoforma corta de endoglina (EngS) como un marcador de senescencia en macrófagos, observándose alteraciones en adhesión, supervivencia, proliferación o acumulación de especies reactivas de oxígeno. Los análisis in vivo e in vitro realizados, confirman la validez del desequilibrio entre la expresión de la isoforma corta frente a la mayoritaria (isoforma larga, EngL) como un marcador del estado envejecido en estas células. Además, se ha observado una polarización preferencial de los transfectantes de endoglina corta hacia el fenotipo macrofágico anti-inflamatorio-M2 y una peor respuesta generalizada a estímulos pro-inflamatorios-M1. Este comportamiento se puede englobar dentro del deterioro en la respuesta inmune innata asociada a la inmunosenescencia. Por otra parte se ha analizado el perfil liberación de una tercera forma de endoglina, la endoglina soluble. Se ha identificado por primera vez esta forma soluble de endoglina como un marcador de polarización de macrófagos M1. Además se ha descrito un nuevo mecanismo proteolítico de liberación de endoglina de membrana por medio de la metaloproteasa de macrófagos MMP12. Se ha establecido por tanto una relación directa entre endoglina soluble y MMP12 como marcadores de respuesta inflamatoria en macrófagos, un hallazgo que está apoyado por el hecho de que ambos factores solubles están aumentados en enfermedades que tienen asociado un proceso inflamatorio.
Endoglin is a membrane glycoprotein with a crucial role in vascular remodeling and angiogenesis. Being described its function and pathophysiological implications mainly in the field of endothelial cell, its role in the myeloid lineage, in which is also expressed, is still to be elucidated. In the present memory, the role of the two isoforms of endoglin in the U937 promonocytic cell line by SILAC proteomic analysis is studied. Analyzing the altered proteins due to the action of the isoforms of endoglin, it has been identified the short isoform of endoglin (EngS) as a marker of senescence in macrophages, observing changes in adhesion, survival, proliferation or reactive oxygen species (ROS) accumulation. The performed in vivo and in vitro assays, confirmed the validity of the imbalance between the expression of the short isoform against the majority (long isoform, EngL) as a marker of the aged state in these cells. Furthermore, there has been a preferential polarization of the EngS transfectants into an M2 anti-inflammatory macrophage phenotype and a compromised general response to pro-inflammatory M1 phenotype. This behavior can be included within the impairment of the innate immune response associated with the immunosenescence. Moreover, the releasing profile of a third form of endoglin, soluble endoglin, is analyzed. We have identified for the first time this soluble form of endoglin as an M1 macrophage polarization marker. In addition, a new proteolytic mechanism of membrane endoglin release by macrophage metalloproteinase MMP12 is described. Therefore, it has been established a direct relationship between soluble endoglin and MMP12 as inflammatory markers in macrophages, a finding that is supported by the fact that both soluble factors are increased in diseases associated with an inflammatory process
Endoglin is a membrane glycoprotein with a crucial role in vascular remodeling and angiogenesis. Being described its function and pathophysiological implications mainly in the field of endothelial cell, its role in the myeloid lineage, in which is also expressed, is still to be elucidated. In the present memory, the role of the two isoforms of endoglin in the U937 promonocytic cell line by SILAC proteomic analysis is studied. Analyzing the altered proteins due to the action of the isoforms of endoglin, it has been identified the short isoform of endoglin (EngS) as a marker of senescence in macrophages, observing changes in adhesion, survival, proliferation or reactive oxygen species (ROS) accumulation. The performed in vivo and in vitro assays, confirmed the validity of the imbalance between the expression of the short isoform against the majority (long isoform, EngL) as a marker of the aged state in these cells. Furthermore, there has been a preferential polarization of the EngS transfectants into an M2 anti-inflammatory macrophage phenotype and a compromised general response to pro-inflammatory M1 phenotype. This behavior can be included within the impairment of the innate immune response associated with the immunosenescence. Moreover, the releasing profile of a third form of endoglin, soluble endoglin, is analyzed. We have identified for the first time this soluble form of endoglin as an M1 macrophage polarization marker. In addition, a new proteolytic mechanism of membrane endoglin release by macrophage metalloproteinase MMP12 is described. Therefore, it has been established a direct relationship between soluble endoglin and MMP12 as inflammatory markers in macrophages, a finding that is supported by the fact that both soluble factors are increased in diseases associated with an inflammatory process
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Tesis inédita de la Universidad Complutense de Madrid, Facultad de Ciencias Biológicas, Departamento de Genética, leída el 27-02-2014