Palmdelphin Regulates Nuclear Resilience to Mechanical Stress in the Endothelium
| Autor: | Oscar Plunde, Ross O Smith, Qingsen Li, Marco Foiani, Cansaran Saygili Demir, Anders Franco-Cereceda, Marie Hedlund, Sofia Nordling, Lena Claesson-Welsh, Flora Ascione, Miguel Sáinz-Jaspeado, Yindi Ding, Manfred W. Kilimann, Sven-Christian Pawelzik, Pontus Aspenström, Jeffrey Kroon, Giulia Bastianello, Magnus Bäck, Yi Jin, Tatiana V. Petrova, Dinesh Fernando, Geoffrey Daniel |
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| Přispěvatelé: | Experimental Vascular Medicine, ACS - Atherosclerosis & ischemic syndromes, ACS - Microcirculation, AGEM - Amsterdam Gastroenterology Endocrinology Metabolism |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Male
Gene Expression aortic valve stenosis Cell Communication Mice Cell Movement Original Research Articles Databases Genetic Medicine Aged Animals Cell Communication/genetics Cell Line Cell Movement/genetics Cell Nucleus/genetics Cell Nucleus/metabolism Cells Cultured Computational Biology/methods Endothelial Cells/metabolism Endothelium/metabolism Female Gene Expression Profiling Gene Knockdown Techniques Gene Ontology Humans Immunohistochemistry Membrane Proteins/genetics Membrane Proteins/metabolism Mice Knockout Middle Aged Protein Transport Stress Mechanical endothelial cells nucleocytoplasmic transport palmdelphin Cardiac and Cardiovascular Systems Cytoskeleton Kardiologi Cell biology medicine.anatomical_structure ComputingMethodologies_DOCUMENTANDTEXTPROCESSING Palmdelphin Cardiology and Cardiovascular Medicine Endothelium Locus (genetics) Single-nucleotide polymorphism Paralemmin Physiology (medical) Resilience (network) Cell Nucleus business.industry Computational Biology Membrane Proteins Immunology in the medical area Nucleocytoplasmic Transport business |
| Zdroj: | Circulation, vol. 144, no. 20, pp. 1629-1645 Circulation, 144(20), 1629-1645. Lippincott Williams and Wilkins Circulation |
| ISSN: | 0009-7322 |
| Popis: | Supplemental Digital Content is available in the text. Background: PALMD (palmdelphin) belongs to the family of paralemmin proteins implicated in cytoskeletal regulation. Single nucleotide polymorphisms in the PALMD locus that result in reduced expression are strong risk factors for development of calcific aortic valve stenosis and predict severity of the disease. Methods: Immunodetection and public database screening showed dominant expression of PALMD in endothelial cells (ECs) in brain and cardiovascular tissues including aortic valves. Mass spectrometry, coimmunoprecipitation, and immunofluorescent staining allowed identification of PALMD partners. The consequence of loss of PALMD expression was assessed in small interferring RNA-treated EC cultures, knockout mice, and human valve samples. RNA sequencing of ECs and transcript arrays on valve samples from an aortic valve study cohort including patients with the single nucleotide polymorphism rs7543130 informed about gene regulatory changes. Results: ECs express the cytosolic PALMD-KKVI splice variant, which associated with RANGAP1 (RAN GTP hydrolyase activating protein 1). RANGAP1 regulates the activity of the GTPase RAN and thereby nucleocytoplasmic shuttling via XPO1 (Exportin1). Reduced PALMD expression resulted in subcellular relocalization of RANGAP1 and XPO1, and nuclear arrest of the XPO1 cargoes p53 and p21. This indicates an important role for PALMD in nucleocytoplasmic transport and consequently in gene regulation because of the effect on localization of transcriptional regulators. Changes in EC responsiveness on loss of PALMD expression included failure to form a perinuclear actin cap when exposed to flow, indicating lack of protection against mechanical stress. Loss of the actin cap correlated with misalignment of the nuclear long axis relative to the cell body, observed in PALMD-deficient ECs, Palmd−/− mouse aorta, and human aortic valve samples derived from patients with calcific aortic valve stenosis. In agreement with these changes in EC behavior, gene ontology analysis showed enrichment of nuclear- and cytoskeleton-related terms in PALMD-silenced ECs. Conclusions: We identify RANGAP1 as a PALMD partner in ECs. Disrupting the PALMD/RANGAP1 complex alters the subcellular localization of RANGAP1 and XPO1, and leads to nuclear arrest of the XPO1 cargoes p53 and p21, accompanied by gene regulatory changes and loss of actin-dependent nuclear resilience. Combined, these consequences of reduced PALMD expression provide a mechanistic underpinning for PALMD’s contribution to calcific aortic valve stenosis pathology. |
| Databáze: | OpenAIRE |
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