Acetyl-CoA carboxylase 1 regulates endothelial cell migration by shifting the phospholipid composition

Abstract

The enzyme acetyl-CoA carboxylase (ACC) plays a crucial role in fatty acid metabolism. In recent years, ACC has been recognized as a promising drug target for treating different diseases. However, the role of ACC in vascular endothelial cells (ECs) has been neglected so far. To characterize the role of ACC, we used the ACC inhibitor, soraphen A, as a chemical tool, and also a gene silencing approach. We found that ACC1 was the predominant isoform in human umbilical vein ECs as well as in human microvascular ECs and that soraphen A reduced the levels of malonyl-CoA. We revealed that ACC inhibition shifted the lipid composition of EC membranes. Accordingly, membrane fluidity, filopodia formation, and migratory capacity were reduced. The antimigratory action of soraphen A depended on an increase in the cellular proportion of PUFAs and, most importantly, on a decreased level of phosphatidylglycerol. Our study provides a causal link between ACC, membrane lipid composition, and cell migration in ECs. Soraphen A represents a useful chemical tool to investigate the role of fatty acid metabolism in ECs and ACC inhibition offers a new and valuable therapeutic perspective for the treatment of EC migration-related diseases. The enzyme acetyl-CoA carboxylase (ACC) plays a crucial role in fatty acid metabolism. In recent years, ACC has been recognized as a promising drug target for treating different diseases. However, the role of ACC in vascular endothelial cells (ECs) has been neglected so far. To characterize the role of ACC, we used the ACC inhibitor, soraphen A, as a chemical tool, and also a gene silencing approach. We found that ACC1 was the predominant isoform in human umbilical vein ECs as well as in human microvascular ECs and that soraphen A reduced the levels of malonyl-CoA. We revealed that ACC inhibition shifted the lipid composition of EC membranes. Accordingly, membrane fluidity, filopodia formation, and migratory capacity were reduced. The antimigratory action of soraphen A depended on an increase in the cellular proportion of PUFAs and, most importantly, on a decreased level of phosphatidylglycerol. Our study provides a causal link between ACC, membrane lipid composition, and cell migration in ECs. Soraphen A represents a useful chemical tool to investigate the role of fatty acid metabolism in ECs and ACC inhibition offers a new and valuable therapeutic perspective for the treatment of EC migration-related diseases. The multi-domain enzyme acetyl-CoA carboxylase (ACC) is crucially involved in the fatty acid metabolism of eukaryotes. In the recent years, ACC has been increasingly recognized as a promising drug target for the treatment of different diseases (1.Tong L. Acetyl-coenzyme A carboxylase: crucial metabolic enzyme and attractive target for drug discovery.Cell. Mol. Life Sci. 2005; 62: 1784-1803Crossref PubMed Scopus (366) Google Scholar, 2.Tong L. Harwood Jr, H.J. Acetyl-coenzyme A carboxylases: versatile targets for drug discovery.J. Cell. Biochem. 2006; 99: 1476-1488Crossref PubMed Scopus (163) Google Scholar): i) The persistent increase in the incidence of the metabolic syndrome has aroused renewed interest in ACC after its discovery about 60 years ago (1.Tong L. Acetyl-coenzyme A carboxylase: crucial metabolic enzyme and attractive target for drug discovery.Cell. Mol. Life Sci. 2005; 62: 1784-1803Crossref PubMed Scopus (366) Google Scholar, 3.Wakil S.J. Stoops J.K. Joshi V.C. Fatty acid synthesis and its regulation.Annu. Rev. Biochem. 1983; 52: 537-579Crossref PubMed Google Scholar). ACC inhibition or knockdown promises beneficial effects to control lipid synthesis in an effort to treat metabolic diseases like obesity and diabetes (4.Abu-Elheiga L. Matzuk M.M. Abo-Hashema K.A. Wakil S.J. Continuous fatty acid oxidation and reduced fat storage in mice lacking acetyl-CoA carboxylase 2.Science. 2001; 291: 2613-2616Crossref PubMed Scopus (742) Google Scholar, 5.Schreurs M. Van Dijk T. Gerding A. Havinga R. Reijngoud D.J. Kuipers F. Soraphen, an inhibitor of the acetyl-CoA carboxylase system, improves peripheral insulin sensitivity in mice fed a high-fat diet.Diabetes Obes. Metab. 2009; 11: 987-991Crossref PubMed Scopus (36) Google Scholar, 6.Harwood Jr, H.J. Acetyl-CoA carboxylase inhibition for the treatment of metabolic syndrome.Curr. Opin. Investig. Drugs. 2004; 5: 283-289PubMed Google Scholar, 7.Harwood Jr, H.J. Treating the metabolic syndrome: acetyl-CoA carboxylase inhibition.Expert Opin. Ther. Targets. 2005; 9: 267-281Crossref PubMed Scopus (113) Google Scholar, 8.Stiede K. Miao W. Blanchette H.S. Beysen C. Harriman G. Harwood Jr., H.J. Kelley H. Kapeller R. Schmalbach T. Westlin W.F. Acetyl-CoA carboxylase inhibition reduces de novo lipogenesis in overweight male subjects: a randomized, double-blind, crossover study.Hepatology. 2017; 66: 324-334Crossref PubMed Scopus (82) Google Scholar, 9.Harriman G. Greenwood J. Bhat S. Huang X. Wang R. Paul D. Tong L. Saha A.K. Westlin W.F. Kapeller R. et al.Acetyl-CoA carboxylase inhibition by ND-630 reduces hepatic steatosis, improves insulin sensitivity, and modulates dyslipidemia in rats.Proc. Natl. Acad. Sci. USA. 2016; 113: E1796-E1805Crossref PubMed Scopus (183) Google Scholar). ii) Several studies reported that ACC is upregulated in different human cancers. Because this phenomenon is associated with increased lipogenesis and rapid cancer cell growth, the inhibition of ACC has been suggested as a promising therapeutic strategy (10.Swinnen J.V. Brusselmans K. Verhoeven G. Increased lipogenesis in cancer cells: new players, novel targets.Curr. Opin. Clin. Nutr. Metab. Care. 2006; 9: 358-365Crossref PubMed Scopus (472) Google Scholar, 11.Milgraum L.Z. Witters L.A. Pasternack G.R. Kuhajda F.P. Enzymes of the fatty acid synthesis pathway are highly expressed in in situ breast carcinoma.Clin. Cancer Res. 1997; 3: 2115-2120PubMed Google Scholar, 12.Beckers A. Organe S. Timmermans L. Scheys K. Peeters A. Brusselmans K. Verhoeven G. Swinnen J.V. Chemical inhibition of acetyl-CoA carboxylase induces growth arrest and cytotoxicity selectively in cancer cells.Cancer Res. 2007; 67: 8180-8187Crossref PubMed Scopus (245) Google Scholar, 13.Yahagi N. Shimano H. Hasegawa K. Ohashi K. Matsuzaka T. Najima Y. Sekiya M. Tomita S. Okazaki H. Tamura Y. et al.Co-ordinate activation of lipogenic enzymes in hepatocellular carcinoma.Eur. J. Cancer. 2005; 41: 1316-1322Abstract Full Text Full Text PDF PubMed Scopus (190) Google Scholar). iii) Furthermore, both the pharmacological inhibition and the deletion of ACC1 impaired TH17 cell-mediated autoimmune disease in mice (14.Berod L. Friedrich C. Nandan A. Freitag J. Hagemann S. Harmrolfs K. Sandouk A. Hesse C. Castro C.N. Bahre H. et al.De novo fatty acid synthesis controls the fate between regulatory T and T helper 17 cells.Nat. Med. 2014; 20: 1327-1333Crossref PubMed Scopus (512) Google Scholar). ACC catalyzes the carboxylation of acetyl-CoA into malonyl-CoA. This reaction is the first and rate-limiting step in the biosynthesis of fatty acids in all organisms (7.Harwood Jr, H.J. Treating the metabolic syndrome: acetyl-CoA carboxylase inhibition.Expert Opin. Ther. Targets. 2005; 9: 267-281Crossref PubMed Scopus (113) Google Scholar, 15.Kim K-H. Regulation of mammalian acetyl-coenzyme A carboxylase.Annu. Rev. Nutr. 1997; 17: 77-99Crossref PubMed Scopus (308) Google Scholar, 16.McGarry J.D. Mannaerts G. Foster D.W. A possible role for malonyl-CoA in the regulation of hepatic fatty acid oxidation and ketogenesis.J. Clin. Invest. 1977; 60: 265-270Crossref PubMed Scopus (514) Google Scholar). Malonyl-CoA is an essential substrate for fatty acid synthesis and chain elongation and functions as a strong inhibitor of mitochondrial fatty acid β-oxidation. There are two tissue-specific isoforms of ACC in mammalian cells: ACC1, present in lipogenic tissue (liver, adipose), and ACC2, present in oxidative tissues (liver, heart, skeletal muscle) (1.Tong L. Acetyl-coenzyme A carboxylase: crucial metabolic enzyme and attractive target for drug discovery.Cell. Mol. Life Sci. 2005; 62: 1784-1803Crossref PubMed Scopus (366) Google Scholar). Cytosolic ACC1 generates malonyl-CoA for de novo lipogenesis, while mitochondrial ACC2 generates malonyl-CoA that acts as an inhibitor of carnitine palmitoyltransferase. This enzyme transfers fatty acids into the mitochondria for β-oxidation to acetyl-CoA (16.McGarry J.D. Mannaerts G. Foster D.W. A possible role for malonyl-CoA in the regulation of hepatic fatty acid oxidation and ketogenesis.J. Clin. Invest. 1977; 60: 265-270Crossref PubMed Scopus (514) Google Scholar, 17.Saggerson D. Malonyl-CoA, a key signaling molecule in mammalian cells.Annu. Rev. Nutr. 2008; 28: 253-272Crossref PubMed Scopus (198) Google Scholar). Despite the great interest in ACC as pharmacological drug target, no attention has as yet been given to the role of ACC in vascular endothelial cells (ECs). A monolayer of ECs forms the inner lining of each blood vessel and provides a barrier that tightly controls the passage of fluid, solutes, macromolecules, and cells from the blood to the underlying tissue (18.Sumpio B.E. Riley J.T. Dardik A. Cells in focus: endothelial cell.Int. J. Biochem. PubMed Scopus Google Scholar, J.D. endothelial cell 9: PubMed Scopus Google Scholar). its as a the vascular also as a and the J.D. endothelial cell 9: PubMed Scopus Google Scholar). A of ECs is to for and growth as well as and tissue its as a EC migration is of the and of as and cancer L. F. J. cell migration Res. 2007; PubMed Scopus Google Scholar, of endothelial cell Mol. Life Sci. 2014; PubMed Scopus Google Scholar). with EC migration offers an therapeutic approach. the of ACC for the migration of ECs has been neglected so far. this study to investigate the role of ACC in human ECs with a on membrane and the migratory capacity of ECs. To the of ACC, we used the ACC inhibitor, soraphen A, a from the as well as a gene approach. Soraphen A is to ACC by to the carboxylase of ACC1 and The of soraphen A with the of this is for ACC Y. Tong L. A for the inhibition of acetyl-coenzyme A carboxylase by soraphen A, a Cell. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar, D. for the regulation of human ACC2 by Res. PubMed Scopus Google Scholar). Our for the first that ACC1 endothelial filopodia and, most importantly, EC migration by the The present study a in the by the between the fatty acid metabolism and the migration of ECs. Soraphen A was by et K. S. S. 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PubMed Scopus Google Scholar). as well as levels of ACC1 and ACC2 were by and ACC1 and ACC2 levels in and were with the human cancer cell by and that the ACC1 in was about of the in cells and the in was about of the in In the of ACC2 in both and was of the levels found in cells Accordingly, by that the ACC1 in was about the of ACC2 The of the were in with that ACC1 is the predominant isoform in ECs. Several studies reported that soraphen A is a inhibitor of ACC L. H. A. of the ACC1 gene as the target of the soraphen PubMed Scopus Google Scholar, K. N. H. G. H. The a of novel from PubMed Scopus Google Scholar). To the of soraphen A in we the of malonyl-CoA after soraphen A treatment by Malonyl-CoA levels were reduced soraphen A treatment by Soraphen A the effects in after or of as by the metabolic the with and the of cells with membrane of that soraphen A ACC and the of ECs. 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H. A. on the role of lipid synthesis in cancer metabolism and PubMed Scopus Google Scholar, C. L. in cancer J. 2014; PubMed Scopus Google Scholar). with cells fatty acids in an on lipogenesis H. X. and of fatty acid an therapeutic target and J. Biochem. Mol. Google Scholar). the fatty acid metabolism of the has ECs are to fatty acids from the de novo lipogenesis has been suggested to of for the and regulation of as well as endothelial as endothelial X. A. novo lipogenesis vascular endothelial Full Text Full Text PDF PubMed Scopus Google endothelial F. D. F. G. L. et metabolic that fatty acid oxidation controls endothelial cell Cell. Full Text Full Text PDF PubMed Scopus Google or and S. K. M. S. A. A. J. et and of by reduces Metab. 2014; Full Text Full Text PDF PubMed Scopus Google Scholar). The inhibition of ACC offers two attractive to with fatty acid the fatty acid oxidation in oxidative tissues and, the de novo lipogenesis in lipogenic tissues L. Harwood Jr, H.J. Acetyl-coenzyme A carboxylases: versatile targets for drug discovery.J. Cell. Biochem. 2006; 99: 1476-1488Crossref PubMed Scopus (163) Google Scholar, S.J. L.A. Fatty acid target for metabolic Res. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar). studies ACC as a valuable drug target for the treatment of the metabolic syndrome and and, of also autoimmune diseases. Harriman et G. Greenwood J. Bhat S. Huang X. Wang R. Paul D. Tong L. Saha A.K. Westlin W.F. Kapeller R. et al.Acetyl-CoA carboxylase inhibition by ND-630 reduces hepatic steatosis, improves insulin sensitivity, and modulates dyslipidemia in rats.Proc. Natl. Acad. Sci. USA. 2016; 113: E1796-E1805Crossref PubMed Scopus (183) Google reported that the ACC inhibitor was to in a of reduced hepatic and and insulin sensitivity and In an in of the ACC inhibitor in growth S.J. M. Van A. L. et of acetyl-CoA carboxylase fatty acid synthesis and growth of cancer in Med. 2016; PubMed Scopus Google Scholar). soraphen A decreased the of TH17 the of and the in a of (14.Berod L. Friedrich C. Nandan A. Freitag J. Hagemann S. Harmrolfs K. Sandouk A. Hesse C. Castro C.N. Bahre H. et al.De novo fatty acid synthesis controls the fate between regulatory T and T helper 17 cells.Nat. Med. 2014; 20: 1327-1333Crossref PubMed Scopus (512) Google Scholar). Despite this the role of ACC and the of ACC inhibition on EC functions been so far. Our study provides the first link between acid metabolism and migration in ECs. 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A. of membrane on cancer cell migration and J. 17: Scopus Google also found that soraphen A treatment of cancer cells increased the of and the chain of fatty membrane and the migratory capacity of In to ACC inhibition to in the of This to the of different cell treatment and of soraphen A. A of study is that as well as PUFAs are of membrane and migration of ECs. PUFAs the antimigratory action of soraphen A. fatty acids the of soraphen A on EC an of the of ACC This that the antimigratory action of soraphen A is to the of present in EC membranes. The role of in cell migration was in the so far. Our study for the first a link between migration and an composition in ECs. the soraphen A represents a useful chemical tool to investigate the role of fatty acid metabolism in ECs. the the inhibition of ACC offers a new and valuable therapeutic perspective for the treatment of EC migration-related diseases. 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