Nathalie Bonello-Palot1, Stéphanie Simoncini2, Stéphane Robert2, Patrice Bourgeois3, Florence Sabatier4, Nicolas Levy1, Françoise Dignat-George5, Catherine Badens6. 1. APHM, CHU de la Timone, Laboratoire de Génétique Moléculaire, Marseille, France; Aix-Marseille Université, Inserm UMR_S U910, Faculté de Médecine, Marseille, France. 2. Aix-Marseille Université, Inserm, VRCM, UMR_S 1076,13005, Marseille, France. 3. APHM, CHU de la Timone, Laboratoire de Génétique Moléculaire, Marseille, France. 4. Aix-Marseille Université, Inserm, VRCM, UMR_S 1076,13005, Marseille, France; APHM, CHU de la Conception, Laboratoire de thérapie cellulaire, Marseille, France. 5. Aix-Marseille Université, Inserm, VRCM, UMR_S 1076,13005, Marseille, France; AP-HM, CHU de la Conception, Laboratoire d'Hématologie-Immunologie, Marseille, France. 6. APHM, CHU de la Timone, Laboratoire de Génétique Moléculaire, Marseille, France; Aix-Marseille Université, Inserm UMR_S U910, Faculté de Médecine, Marseille, France. Electronic address: catherine.badens@univ-amu.fr.
Abstract
BACKGROUND: Defects in lamin A maturation result in premature aging syndromes and severe atherosclerosis as observed in the Hutchinson-Gilford Progeria Syndrome. In age-related atherosclerosis, several features of cellular senescence have been characterized in endothelial cells including telomere shortening and increased oxidative stress. However, to date, very little is known about lamin A alterations in these cells. OBJECTIVES: To study lamin A-related senescence and its consequences in the activation status of primary endothelial cells. METHODS: Healthy primary endothelial cells and progenitors issued from human umbilical vein or cord blood were used. Lamin A defects were induced by protease inhibitor (Atazanavir) treatment for 48 h. RESULTS: We show that protease inhibitor treatment leads to the accumulation of farnesylated prelamin A, inducing nuclear shape abnormalities and premature senescence in both differentiated and progenitor endothelial cells. ICAM-1-dependent activation and monocytes adhesion was increased in mature endothelial cells. In parallel, the ability to generate microvascular networks in matrigel was decreased for endothelial progenitors. The effects of protease inhibitor treatment on nuclear shapes were reversed when cells were treated in combination with Pravastatin and Zoledronate in both mature and progenitor endothelial cells. Reversion was also demonstrated with a morpholino antisense-oligonucleotide targeting lamin A-specific splice site. DISCUSSION: This study shows that protease inhibitor treatment reproduces premature senescence due to lamin A defects in primary endothelial cells and progenitors after 48 h exposure. The cells used were non-aged as extracted from cord blood or umbilical vein, allowing one to consider that other senescence pathways were not activated and that the observed alterations were specific of prelamin A accumulation. Both mature endothelial cells and precursors were sensitive to prelamin accumulation and thus, could be used in the future as a valuable model to test different approaches aimed at specifically reversing lamin A-related cells senescence.
BACKGROUND: Defects in lamin A maturation result in premature aging syndromes and severe atherosclerosis as observed in the Hutchinson-Gilford Progeria Syndrome. In age-related atherosclerosis, several features of cellular senescence have been characterized in endothelial cells including telomere shortening and increased oxidative stress. However, to date, very little is known about lamin A alterations in these cells. OBJECTIVES: To study lamin A-related senescence and its consequences in the activation status of primary endothelial cells. METHODS: Healthy primary endothelial cells and progenitors issued from human umbilical vein or cord blood were used. Lamin A defects were induced by protease inhibitor (Atazanavir) treatment for 48 h. RESULTS: We show that protease inhibitor treatment leads to the accumulation of farnesylated prelamin A, inducing nuclear shape abnormalities and premature senescence in both differentiated and progenitor endothelial cells. ICAM-1-dependent activation and monocytes adhesion was increased in mature endothelial cells. In parallel, the ability to generate microvascular networks in matrigel was decreased for endothelial progenitors. The effects of protease inhibitor treatment on nuclear shapes were reversed when cells were treated in combination with Pravastatin and Zoledronate in both mature and progenitor endothelial cells. Reversion was also demonstrated with a morpholino antisense-oligonucleotide targeting lamin A-specific splice site. DISCUSSION: This study shows that protease inhibitor treatment reproduces premature senescence due to lamin A defects in primary endothelial cells and progenitors after 48 h exposure. The cells used were non-aged as extracted from cord blood or umbilical vein, allowing one to consider that other senescence pathways were not activated and that the observed alterations were specific of prelamin A accumulation. Both mature endothelial cells and precursors were sensitive to prelamin accumulation and thus, could be used in the future as a valuable model to test different approaches aimed at specifically reversing lamin A-related cells senescence.
Authors: Gianfranco Matrone; Rajarajan A Thandavarayan; Brandon K Walther; Shu Meng; Anahita Mojiri; John P Cooke Journal: Cell Cycle Date: 2019-08-14 Impact factor: 4.534
Authors: Nazish Sayed; Chun Liu; Mohamed Ameen; Farhan Himmati; Joe Z Zhang; Saereh Khanamiri; Jan-Renier Moonen; Alexa Wnorowski; Linling Cheng; June-Wha Rhee; Sadhana Gaddam; Kevin C Wang; Karim Sallam; Jack H Boyd; Y Joseph Woo; Marlene Rabinovitch; Joseph C Wu Journal: Sci Transl Med Date: 2020-07-29 Impact factor: 17.956