Low-density lipoprotein receptor-related protein 1 mediates hypoxia-induced very low density lipoprotein-cholesteryl ester uptake and accumulation in cardiomyocytes

Cal, Roi; Castellano, José; Revuelta‐López, Elena; Aledo, Rosa; Barriga, Montse; Farré, Jordi; Vilahur, Gemma; Nasarre, Laura; Hove‐Madsen, Leif; Badimon, Lina; Llorente‐Cortés, Vicenta

doi:10.1093/cvr/cvs136

Cited by 54 publications

(81 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We previously showed that hypoxia-induced LRP1 overexpression is mediated by HIF-1α in hVSMCs and HL-1 cells. 16,17 HIF-1α expression depends on intracellular calcium concentrations that rapidly and persistently increase in response to hypoxia. 31,32 Hypoxia-induced Pyk2 phosphorylation has been associated to a hypoxia-induced increase in calcium cytosolic levels.…”

Section: Discussionmentioning

confidence: 99%

Hypoxia Induces Metalloproteinase-9 Activation and Human Vascular Smooth Muscle Cell Migration Through Low-Density Lipoprotein Receptor–Related Protein 1–Mediated Pyk2 Phosphorylation

Revuelta‐López

Castellano

Roura

et al. 2013

ATVB

Self Cite

View full text Add to dashboard Cite

Objective— Hypoxia disturbs vascular function by promoting extracellular matrix remodeling. Extracellular matrix integrity and composition are modulated by metalloproteinases (MMPs). Our aim was to investigate the role of low-density lipoprotein receptor–related protein 1 (LRP1) in regulating MMP-9/MMP-2 activation and vascular smooth muscle cells (VSMCs) migration in response to hypoxia, and to elucidate the LRP1-signaling pathways involved in this process. Approach and Results— Western blot analysis showed that hypoxia induced a sustained phosphorylation of proline-rich tyrosine kinase 2 concomitantly with LRP1 overexpression in human VSMCs (hVSMCs). Deletion of LRP1 using small-interfering RNA technology or treatment of hVSMCs with the Src family kinase inhibitor PP2 impaired hypoxia-induced phosphorylation of proline-rich tyrosine kinase 2 levels. Coimmunoprecipitation experiments showed that the higher amounts of phosphorylation of proline-rich tyrosine kinase 2/LRP1β immunoprecipitates in hypoxic hVSMCs were abolished in PP2-treated hVSMCs. Both LRP1 silencing and PP2 treatment were highly effective in the prevention of hypoxia-induced MMP-9 activation and hVSMC migration. Cellular subfractionation experiments revealed that PP2 effects may be caused by impairment of hypoxia-induced nuclear factor-κβ translocation to the nucleus. ELISA measurements showed that LRP1 silencing but not PP2 treatment increased interleukin-1β, interleukin-6, and monocyte chemoattractant protein-1 secretion by hypoxic hVSMCs. Conclusions— Our findings determine a crucial role of LRP1-mediated Pyk2 phosphorylation on hypoxia-induced MMP-9 activation and hVSMC migration and therefore in hypoxia-induced vascular remodeling. Both LRP1 silencing and PP2 treatments also influence hypoxia-induced proinflammatory effects in hVSMCs. Therefore, further studies are required to establish therapeutical strategies that efficiently modulate vascular remodeling and inflammation associated with hypoxia-vascular diseases.

show abstract

Section: Discussionmentioning

confidence: 99%

Hypoxia Induces Metalloproteinase-9 Activation and Human Vascular Smooth Muscle Cell Migration Through Low-Density Lipoprotein Receptor–Related Protein 1–Mediated Pyk2 Phosphorylation

Revuelta‐López

Castellano

Roura

et al. 2013

ATVB

Self Cite

View full text Add to dashboard Cite

show abstract

“…Human VSMC were preexposed to agLDL (100 μg/mL) for 18 hours before addition of cycloheximide (Sigma, 100 μmol/L). Cells were harvested at various time points after cycloheximide treatment (6,12,24, and 32 hours) and collected in lysis buffer, and protein processed for Western blot analysis. LRP1 protein stability was assessed as the proportion of the initial protein remaining after cycloheximide treatment.…”

Section: Measurement Of Lrp1 Stability After Exposure To Agldlmentioning

confidence: 99%

“…Our group reported that LRP1 is upregulated at transcriptional level by hypercholesterolemia through sterol regulatory element-binding proteins (SREBP) downregulation in human VSMC, [8][9][10] and by hypoxia through hypoxia-inducible factor 1α upregulation in human VSMC 11 and cardiomyocytes. 12,13 Other groups have reported that insulin promotes the presence of LRP1 in the plasma membrane without influencing mRNA expression levels. 14,15 LRP1 cellular turnover is regulated by the ubiquitinproteasome system, 16 an important mechanism for targeting membrane proteins to destruction, and therefore key in the modulation of protein levels.…”

mentioning

confidence: 99%

Aggregated Low-Density Lipoprotein Induces LRP1 Stabilization Through E3 Ubiquitin Ligase CHFR Downregulation in Human Vascular Smooth Muscle Cells

Cal

García-Arguinzonis

Revuelta‐López

et al. 2013

ATVB

Self Cite

View full text Add to dashboard Cite

Objective-Low density lipoprotein retention and aggregation in the arterial intima are key processes in atherogenesis. Aggregated LDL (agLDL) is taken up through low-density lipoprotein receptor-related protein 1 (LRP1) by human vascular smooth muscle cells (VSMC). AgLDL increases LRP1 expression, at least in part, by downregulation of sterol regulatory element-binding proteins. It is unknown whether agLDL has some effect on the ubiquitin-proteasome system, and therefore on the LRP1 receptor turnover. The objective of this study was to analyze the effect of agLDL on the degradation of LRP1 by the ubiquitin-proteasome system in human VSMC. Methods and Results-Human VSMC were isolated from the media of human coronary arteries. Ubiquitinylated LRP1 protein levels were significantly reduced in human VSMC exposed to agLDL (100 μg/mL) for 20 hours (agLDL: 3.70±0.44 a.u. versus control: 9.68±0.55 a.u). Studies performed with cycloheximide showed that agLDL prolongs the LRP1 protein half life. Pulse-chase analysis showed that LRP1 turnover rate is reduced in agLDL-exposed VSMC. Two-dimensional electrophoresis shows an alteration in the proteomic profile of a RING type E3 ubiquitin ligase, CHFR. Real-time PCR and Western blot analysis showed that agLDL (100 μg/mL) decreased the transcriptional and protein expression of CHFR. CHFR silencing increased VSMC, but not macrophage, LRP1 expression. However, CHFR silencing did not exert any effect on the classical low-density lipoprotein receptor protein levels. Furthermore, immunoprecipitation experiments demonstrated that the physical interaction between CHFR and LRP1 decreased in the presence of agLDL. Conclusion-Our results demonstrate that agLDL prolongs the half life of LRP1 by preventing the receptor ubiquitinylation, at least in part, through CHFR targeting. This mechanism seems to be specific for LRP1 and VSMC.

show abstract

“…Here, we also show that one of the mechanisms leading to higher intracellular TG accumulation under hypoxia is the increased uptake of LDL(+) and LDL(−) by cardiomyocytes exposed to hypoxic conditions. We have previously reported that the classical LDL receptor recognizes LDL(−) with a lower affinity than native LDL , and that this receptor is downregulated in hypoxic cardiomyocytes (Cal et al, 2012a). These observations suggest that alternative receptors whose expression is upregulated by hypoxia may be involved.…”

Section: Discussionmentioning

confidence: 90%

“…In addition, LRP1 internalizes aggregated LDL (Llorente-Cortes et al, 2000). Moreover, hypoxia upregulates the expression of CD36 (Mwaikambo et al, 2009), LOX-1 (Crucet et al, 2013) and LRP1 (Castellano et al, 2011a, Cal et al, 2012a. Thus, one or several of these receptors could be involved in the increased uptake of LDL(−) by cardiomyocytes.…”

Section: Discussionmentioning

confidence: 99%

Hypoxia worsens the impact of intracellular triglyceride accumulation promoted by electronegative low-density lipoprotein in cardiomyocytes by impairing perilipin 5 upregulation

Revuelta‐López

Cal

Julve

et al. 2015

The International Journal of Biochemistry & Cell Biology

Self Cite

View full text Add to dashboard Cite

Low-density lipoprotein receptor-related protein 1 mediates hypoxia-induced very low density lipoprotein-cholesteryl ester uptake and accumulation in cardiomyocytes

Abstract: Our results demonstrate that hypoxia increases LRP1 expression through HIF-1α and that LRP1 overexpression mediates hypoxia-induced VLDL-CE uptake and accumulation in cardiomyocytes.

Cited by 54 publications

References 39 publications

Hypoxia Induces Metalloproteinase-9 Activation and Human Vascular Smooth Muscle Cell Migration Through Low-Density Lipoprotein Receptor–Related Protein 1–Mediated Pyk2 Phosphorylation

Hypoxia Induces Metalloproteinase-9 Activation and Human Vascular Smooth Muscle Cell Migration Through Low-Density Lipoprotein Receptor–Related Protein 1–Mediated Pyk2 Phosphorylation

Aggregated Low-Density Lipoprotein Induces LRP1 Stabilization Through E3 Ubiquitin Ligase CHFR Downregulation in Human Vascular Smooth Muscle Cells

Hypoxia worsens the impact of intracellular triglyceride accumulation promoted by electronegative low-density lipoprotein in cardiomyocytes by impairing perilipin 5 upregulation

Contact Info

Product

Resources

About