Biol. Rev. (2016), 91, pp. 452–468. 452 doi: 10.1111/brv.12178 De novo lipogenesis in the liver in health and disease: more than just a shunting yard for glucose Francis W. B. Sanders1,2 and Julian L. Griffin1,2,∗ 1MRC Human Nutrition Research, Elsie Widdowson Laboratory, 120 Fulbourn Road, Cambridge CB1 9NL, U.K. 2The Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1GA, U.K. ABSTRACT Hepatic de novo lipogenesis (DNL) is the biochemical process of synthesising fatty acids from acetyl-CoA subunits that are produced from a number of different pathways within the cell, most commonly carbohydrate catabolism. In addition to glucose which most commonly supplies carbon units for DNL, fructose is also a profoundly lipogenic substrate that can drive DNL, important when considering the increasing use of fructose in corn syrup as a sweetener. In the context of disease, DNL is thought to contribute to the pathogenesis of non-alcoholic fatty liver disease, a common condition often associated with the metabolic syndrome and consequent insulin resistance. Whether DNL plays a significant role in the pathogenesis of insulin resistance is yet to be fully elucidated, but it may be that the prevalent products of this synthetic process induce some aspect of hepatic insulin resistance. Key words: de novo lipogenesis (DNL), non-alcoholic fatty liver disease (NAFLD), fructose, liver, selective insulin resistance. CONTENTS I. Introduction .............................................................................................. 453 II. The biochemical process of DNL ........................................................................ 453 III. VLDL production and assembly ......................................................................... 455 IV. The regulation of hepatic DNL .......................................................................... 455 (1) SREBP1c ............................................................................................. 455 (2) ChREBP ............................................................................................. 457 (3) ACC .................................................................................................. 458 V. DNL: A Contributor to NAFLD ......................................................................... 460 VI. NAFLD as a component of metabolic syndrome and type 2 diabetes mellitus ........................... 460 VII. Selective insulin resistance promotes hepatic DNL ....................................................... 461 VIII. Anatomical zonation of the metabolic processes in the liver may underlie ‘selective’ insulin resistance ... 462 IX. Fructose, DNL and hepatic steatosis ..................................................................... 463 X. Fructose and the gut microbiome ........................................................................ 463 XI. Conclusions .............................................................................................. 464 XII. Acknowledgements ....................................................................................... 464 XIII. References ................................................................................................ 464 * Address for correspondence (Tel: 01223 437503; E-mail: Jules.griffi[email protected]). Biological Reviews 91 (2016) 452–468 © 2015 The Authors. Biological Reviews published by John Wiley & Sons Ltd on behalf of Cambridge Philosophical Society. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. Hepatic de novo lipogenesis and metabolic disease 453 I. INTRODUCTION Initially this is an acetyl unit bound to the thiol group of cysteine (Cys161)attheβ-ketoacyl synthase (KS) active Hepatic de novo lipogenesis (DNL) is a fundamental site (Witkowski, Joshi & Smith, 1997). The malonyl moiety biosynthetic pathway within the liver, contributing to undergoes decarboxylative condensation with an acetyl the lipids that are stored and secreted by hepatocytes moiety, as elucidated in Fig. 1 (von Wettstein-Knowles et al., (Jensen-Urstad & Semenkovich, 2012). This process is an 2006). ACP is then bound to a β-ketoacyl intermediate. extension of the complex metabolic networks at play within ACP shuttles the β-ketoacyl intermediate to the the liver, and is provided with substrate primarily through NADPH-dependent β-ketoreductase (KR) active site glycolysis and the metabolism of carbohydrates. Therefore, a (Chang & Hammes, 1990). The ketone of the β-carbon high-carbohydrate diet can prime the DNL pathway with a is reduced, generating a hydroxyl group. This is followed large substrate load and increase rates of DNL (Schwarz et al., by sequential dehydration, by the dehydratase (DH) 2003). Importantly this leads to an accumulation of DNL active site, and further reduction by NADPH-dependent products, fatty acyl chains linked to coenzyme A, which can enoyl-reductase (EnR) (Wakil, 1989; Chang & Hammes, be incorporated into a plethora of lipid species. These lipids 1990; Smith & Tsai, 2007). This generates a saturated acyl may then have further metabolic functions, which in turn chain elongated by two carbon groups, which can act as may be deleterious in cases of elevated DNL. the substrate for the next round of elongation as it binds the DNL has been suggested to be abnormally increased in thiol-group of the cysteine at the catalytic site of KS. and contribute to the pathogenesis of non-alcoholic fatty liver The elongation ceases at the 16- or 18-carbon stage disease (NAFLD) (Donnelly et al., 2005), a highly prevalent (Foster & Bloom, 1963; Carey, Dils & Hansen, 1970) with metabolic disease that is linked to the development of type release of palmitic acid or stearic acid from ACP via activity 2 diabetes mellitus (T2DM). DNL is also increased under of the thioesterase (TE) domain of FAS (Chakravarty et al., conditions of insulin resistance (Ameer et al., 2014), and thus 2004; Smith & Tsai, 2007). The specificity of FAS TE for knowing the rate of DNL in an individual may benefit the 16-carbon acyl dictates the length of the FAs released in vitro clinician as it would provide early warning of the possible as there is a rapid decline of TE activity for chain lengths development of T2DM. This is especially important as recent less than 14-carbons (Lin & Smith, 1978; Chakravarty et al., evidence suggests that ‘prediabetes’, a state of mildly elevated 2004), as it will not access the catalytic core of the domain, blood glucose, has increased significantly from 11.6% in 2003 and greater than 18-carbons as it may not be accommodated to 35.3% in 2011 amongst adults in England (Mainous et al., by the binding groove of TE (Chakravarty et al., 2004). The 2014). Therefore it may be crucial to understand and map termination of chain elongation at this 16- carbon stage is the full spectrum of metabolic disturbances associated with further promoted due to acyl chains of this length or longer insulin resistance, including rates of DNL. not readily transferring to the thiol-group of the active-site cysteine of KS (Witkowski et al., 1997). The incorporation of stable-isotope-labelled precursors into palmitate in humans, 3 and tritium from H2O in rat liver also supports palmitate II. THE BIOCHEMICAL PROCESS OF DNL as the major product of DNL (Foster & Bloom, 1963; Hellerstein et al., 1991; Murphy, 2006). DNL is the synthesis of fatty acid (FA) chains from acetyl-CoA While glucose is the main substrate for DNL, fructose subunits produced during glycolysis (Smith & Tsai, 2007) and is a highly lipogenic substrate (Dekker et al., 2010) these can undergo subsequent condensation with a glycerol and this is thought to arise from it bypassing the backbone (Coleman & Lee, 2004) (Fig. 1). critical regulatory step catalysed by phosphofructokinase-1 The reaction mechanism commences with the production (PFK-1) in glycolysis (Basaranoglu, 2013). Fructose is of malonyl-CoA from an acetyl-CoA precursor, under the phosphorylated by fructokinase in the liver to fructose regulated catalytic activity of acetyl-CoA carboxylase (ACC) 1-phosphate (F1P) (Hers, 1952). F1P is then the (Bianchi et al., 1990). The malonyl-CoA is transferred to substrate for catalytic cleavage by aldolase, generating the prosthetic phosphopantetheine group of acyl carrier dihydroxy-acetone-phosphate (DHAP) and glyceraldehyde. protein (ACP) (Majerus, Alberts & Vagelos, 1964), a Glyceraldehyde is subsequently phosphorylated by triokinase domain of the type I fatty acid synthase complex (FAS) to produce glyceraldehyde 3-phosphate (G3P) (Mayes, 1993). (Brindley, Matsumura & Bloch, 1969; Smith, 1994), with Thus G3P and DHAP can enter glycolysis (Fig. 2). This subsequent release of the coenzyme A carrier, catalysed by has implications that mean fructose may reinforce some of the activity of the malonyl/acetyl transferase (MAT) site the pathogenic mechanisms leading to NAFLD (Laville & of mammalian FAS (Mikkelsen et al., 1985). The prosthetic Nazare, 2009; Basaranoglu, 2013), as discussed in Sections phosphopantetheine arm of ACP thus shuttles the elongating IX and X. FA chain to the various catalytic centres in the active site cleft During de novo triacylglycerol (TG) synthesis FAs are of FAS, aided by the rotation of FAS (Wakil, 1989; Smith & incorporated through the initial acylation, by acyl-CoA, Tsai, 2007; Maier, Leibundgut & Ban, 2008). of glycerol-3-phosphate,