RT info:eu-repo/semantics/article
T1 The Integrin Beta1 Modulator Tirofiban Prevents Adipogenesis and Obesity by the Overexpression of Integrin-Linked Kinase: a Pre-Clinical Approach In Vitro and In Vivo
A1 Griera Merino, Mercedes
A1 Hatem Vaquero, Marco Antonio
A1 Campillo de Blas, Sofía
A1 Gutiérrez Calabrés, Elena
A1 García Ayuso, Diego
A1 Pardo, María
A1 Calleros Basilio, Laura
A1 Rodríguez Puyol, Manuel
A1 Rodríguez Puyol, Diego María
A1 Frutos García, Sergio de
K1 Tirofban
K1 Integrin-beta1
K1 Integrin-linked kinase
K1 AKT
K1 Adipogenesis
K1 Obesity
K1 Hypertrophic adipocyte
K1 Browning
K1 White adipose tissue
K1 Lipolysis
K1 Inflammation
K1 Biología y Biomedicina
K1 Biology
AB Background: Obesity is caused by the enlargement of the white adipose tissue (WAT) depots, characterized by the hypertrophic enlargement of malfunctioning adipocytes within WAT which increases the storage of triglycerides (TG) in the lipid droplets (LD). Adipogenesis pathways as well as the expression and activity of some extracellular matrix receptors integrins are upregulated. Integrin?1 (INTB1) is the main isoform involved in WAT remodeling during obesity and insulin resistance-related diseases. We recently described Integrin Linked Kinase (ILK), a scafold protein recruited by INTB1, as an important mediator of WAT remodeling and insulin resistance. As the few approved drugs to fght obesity have brought long-term cardiovascular side efects and given that the consideration of INTB1 and/or ILK modulation as anti-obesogenic strategies remains unexplored, we aimed to evaluate the anti-obesogenic capacity of the clinically approved anticoagulant Tirofban (TF), stated in preclinical studies as a cardiovascular protector. Methods: Fully diferentiated adipocytes originating from C3H10T1/2 were exposed to TF and were co-treated with specifc INTB1 blockers or with siRNA-based knockdown ILK expression. Lipid-specifc dyes were used to determine the TG content in LD. The genetic expression pattern of ILK, pro-infammatory cytokines (MCP1, IL6), adipogenesis (PPAR?, Leptin), thermogenesis (UCP1), proliferation (PCNA), lipid metabolism (FASN, HSL, ATGL), and metabolite trans  porters (FABP4, FAT, AQP7) were detected using quantitative PCR. Cytoskeletal actin polymerization was detected by confocal microscopy. Immunoblotting was performed to detect INTB1 phosphorylation at Thr788/9 and ILK activity as phosphorylation levels of protein kinase B (AKT) in Ser473 and glycogen synthase kinase 3? (GSK3?) at Ser9. TF was intraperitoneally administered once per day to wildtype and ILK knockdown mice (cKDILK) challenged with a high-fat diet (HFD) or control diet (STD) for 2 weeks. Body and WAT weight gains were compared. The expression of ILK and other markers was determined in the visceral epididymal (epi) and inguinal subcutaneous (sc) WAT. Results: TF reduced TG content and the expression of adipogenesis markers and transporters in adipocytes, while UCP-1 expression was increased and the expression of lipases, cytokines or PCNA was not afected. Mechanistically, TF rapidly increased and faded the intracellular phosphorylation of INTB1 but not AKT or GSK3?. F-actin levels were rapidly decreased, and INTB1 blockade avoided the TF efect. After 24 h, ILK expression and phosphorylation rates of AKT and GSK3? were upregulated, while ILK silencing increased TG content. INTB1 blockade and ILK silencing avoided TF efects on the TG content and the transcriptional expression of PPAR? and UCP1. In HFD-challenged mice, the systemic administration of TF for several days reduced the weight gain on WAT depots. TF reduced adipogenesis and pro-infammatory biomarkers and increased lipolysis markers HSL and FAT in epiWAT from HFD, while increased UCP1 in scWAT. In both WATs, TF upregulated ILK expression and activity, while no changes were observed in other tissues. In HFD-fed cKDILK, the blunted ILK in epiWAT worsened weight gain and avoided the anti-obesogenic efect of in vivo TF administration. Conclusions: ILK downregulation in WAT can be considered a biomarker of obesity establishment. Via an INTB1-ILK axis, TF restores malfunctioning hypertrophied WAT by changing the expression of adipocyte-related genes, increas  ing ILK expression and activity, and reducing TG storage. TF prevents obesity, a property to be added to its anticoagu  lant and cardiovascular protective advantages.
SN 2045-3701
YR 2022
FD 2022-01-28
LK http://hdl.handle.net/10017/50868
UL http://hdl.handle.net/10017/50868
LA eng
NO de Frutos, S., Griera, M., Hatem-Vaquero, M. et al. The integrin beta1 modulator Tirofiban prevents adipogenesis and obesity by the overexpression of integrin-linked kinase: a pre-clinical approach in vitro and in vivo. Cell Biosci 12, 10 (2022)
NO Instituto de Salud Carlos III
DS MINDS@UW
RD 27-abr-2024