AGE-albumin enhances ABCA1 degradation by ubiquitin-proteasome and lysosomal pathways in macrophages

Author(s):Show less -

Iborra, Rodrigo Tallada ^[1] ; Machado-Lima, Adriana ^{[2, 1]} ; Okuda, Ligia Shimabukuro ^[1] ; Pinto, Paula Ramos ^[1] ; Nakandakare, Edna Regina ^[1] ; Machado, Ubiratan Fabres ^[3] ; Correa-Giannella, Maria Lucia ^{[4, 5]} ; Pickford, Russell ^[6] ; Woods, Tom ^[7] ; Brimble, Margaret A. ^[7] ; Rye, Kerry-Anne ^[8] ; Lu, Rui ^[9] ; Yokoyama, Shinji ^[9] ; Passarelli, Marisa ^[1]

Affiliation

^[1] Univ Sao Paulo, Fac Med, Hosp Clin HCFMUSP, Lab Lipides, LIM 10, Av Dr Arnaldo 455, Room 3305, BR-01246000 Sao Paulo, SP - Brazil

^[2] Univ Sao Judas Tadeu, Sao Paulo - Brazil

^[3] Univ Sao Paulo, Inst Biomed Sci, Dept Physiol & Biophys, Sao Paulo - Brazil

^[4] Univ Sao Paulo, Fac Med, Hosp Clin HCFMUSP, Lab Carboidratos & Radioimunoinsaio, LIM 18, Sao Paulo - Brazil

^[5] Univ Nove Julho, Programa posgrad Med, Sao Paulo - Brazil

^[6] Univ New South Wales, Bioanalyt Mass Spectrometry Facil, Sydney, NSW - Australia

^[7] Univ Auckland, Sch Chem Sci, Auckland - New Zealand

^[8] Univ New South Wales, Fac Med, Sch Med Sci, Lipid Res Grp, Sydney, NSW - Australia

^[9] Chubu Univ, Nutr Hlth Sci Res Ctr, Kasugai, Aichi - Japan

• Document type: Journal article
• Source: JOURNAL OF DIABETES AND ITS COMPLICATIONS; v. 32, n. 1, p. 1-10, JAN 2018.
• Web of Science Citations: 2

Abstract

Background and aims: Advanced glycation end products (AGEs) induce cellular oxidative/endoplasmic reticulum stress and inflammation. We investigated its underlying mechanisms for atherogenesis focusing on regulation of ABCA1 protein decay in macrophages. Methods: The ABCA1 decay rate was evaluated in macrophages after treatment with LXR agonist and by incubation with control (C) or AGE-albumin concomitant or not with cycloheximide, MG-132, ammonium chloride and calpain inhibitors were utilized to inhibit, respectively, proteasome, lysosome and ABCA1 proteolysis at cell surface. ABCA1 was determined by immunoblot and the protein decay rate calculated along time by the slope of the linear regression. Ubiquitination level was determined in ABCA1 immunoprecipitated from whole cell lysate or bulk cell membrane. AGE effect was also analyzed in THP-1 cells transfected with siRNA-RAGE. Carboxymethyllysine (CML) and pyrraline (PYR) were determined by LC/MS. One-way ANOVA and Student t test were utilized to compare results. Results: CML and PYR-albumin were higher in AGE-albumin as compared to C. AGE-albumin reduced ABCA1 in J774 and THP-1 macrophages (20-30%) and induced a higher ABCA1 ubiquitination and a faster protein decay rate that was dependent on the presence of AGE during the kinetics of measurement in the presence of cycloheximide. Proteasomal inhibition restored and lysosomal inhibition partially recovered ABCA1 in cells treated with AGE-albumin. Calpain inhibition was not able to rescue ABCA1. RAGE knockdown prevented the reduction in ABCA1 elicited by AGE. Conclusions: AGE-albumin.diminishes ABCA1 by accelerating its degradation through the proteasomal and lysosomal systems. This may increase lipid accumulation in macrophages by diminishing cholesterol efflux via RAGE signaling contributing to atherosclerosis in diabetes mellitus. (C) 2017 Elsevier Inc. All rights reserved. (AU)

• FAPESP's process: 12/19112-0 - Involvement of the RAGE/AGE axis in macrophage lipid accumulation induced by human advanced glycated albumin - contribution of glycemic control in diabetes mellitus.
• Grantee: Adriana Machado Saldiba de Lima
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 13/26256-1 - Intracellular trafficking of HDL receptor - ABCA-1 - in macrophages treated with advanced glycated albumin: influence of glycemic control in reverse cholesterol transport
• Grantee: Rodrigo Tallada Iborra
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 16/15603-0 - Unraveling mechanisms of glycemic control and chronic complications of Diabetes mellitus: contributions to human health
• Grantee: Ubiratan Fabres Machado
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 12/12088-7 - Glycemic control and the removal of macrophage cholesterol by ABCA-1: role of modified albumin by advanced glycation
• Grantee: Rodrigo Tallada Iborra
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 13/02854-7 - Anti-inflammatory role of apolipoprotein A-IV in diabetes mellitus and its impact on macrophage reverse cholesterol transport: influence of advanced glycation
• Grantee: Ligia Shimabukuro Okuda
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 15/21072-5 - Influence of the glycemic control and nephropathy stage on the reverse cholesterol transport in Diabetes mellitus: role of metabolic memory induced by advanced glycated albumin
• Grantee: Marisa Passarelli
• Support Opportunities: Regular Research Grants