REVIEWS

M E TA B O L I C S I G N A L L I N G

Cytosolic lipolysis and lipophagy:

two sides of the same coin
Rudolf Zechner1,2,, Frank Madeo1,2 and Dagmar Kratky1,3
Abstract | Fatty acids are the most efficient substrates for energy production in vertebrates
and are essential components of the lipids that form biological membranes. Synthesis of
triacylglycerols from non-esterified free fatty acids (FFAs) combined with triacylglycerol storage
represents a highly efficient strategy to stockpile FFAs in cells and prevent FFA-induced
lipotoxicity. Although essentially all vertebrate cells have some capacity to store and utilize
triacylglycerols, white adipose tissue is by far the largest triacylglycerol depot and is uniquely
able to supply FFAs to other tissues. The release of FFAs from triacylglycerols requires their
enzymatic hydrolysis by a process called lipolysis. Recent discoveries thoroughly altered and
extended our understanding of lipolysis. This Review discusses how cytosolic ‘neutral’ lipolysis
and lipophagy, which utilizes ‘acid’ lipolysis in lysosomes, degrade cellular triacylglycerols as well
as how these pathways communicate, how they affect lipid metabolism and energy homeostasis
and how their dysfunction affects the pathogenesis of metabolic diseases. Answers to these
questions will likely uncover novel strategies for the treatment of prevalent metabolic diseases.

Fatty acids
Monocarboxylic acids with
long saturated or unsaturated
aliphatic carbon chains. They
are either unesterified, in free
form (free fatty acids) or
esterified to various alcohols.
Acid–base homeostasis
Maintenance of a constant pH
in extracellular body fluids by
balanced concentrations of
acids and bases.
1
BioTechMed-Graz,
Mozartgasse 12, 8010 Graz,
Austria.
2
Institute of Molecular
Biosciences, University of
Graz, Heinrichstrasse 31,
8010 Graz, Austria.
3
Institute of Molecular
Biology and Biochemistry,
Medical University of Graz,
Neue Stiftingtalstrasse 6/6,
8010 Graz, Austria.
Correspondence to R.Z.
rudolf.zechner@uni-graz.at
doi:10.1038/nrm.2017.76
Published online 30 Aug 2017
Fatty acids are pivotal for life. They constitute integral
parts of biological membranes, harbour the highest
energy density of all known energy substrates, are precur­
sors for numerous lipid mediators and modulate protein
function through posttranslational acylation of target
proteins. Therefore, it is not surprising that fatty acids
in dietary fats and oils are key nutritional components,
accounting for 20–35% of the daily calorie intake in a
normal human diet. Additionally, all other major diet­
ary components, such as monosaccharides (which are
derived from dietary carbohydrates) and amino acids
(which are derived from diet­ary protein), can be con­
verted into free fatty acids (FFAs). Despite their role in
essential cellular functions, high concentrations of FFAs
are toxic because of their limited solubility and amphi­
pathic nature; their adverse impact on cellular acid–base
homeostasis; and their ready transformation into highly
bioactive, cytotoxic lipid species. These destructive effects
of FFAs — collectively referred to as ‘lipotoxicity’ — can
lead to cellular d­ ysfunction and cell death1.
To avoid lipotoxicity, FFAs are detoxified by their three­
fold esterification to the trivalent alcohol glycerol to form
triacylglycerols. Triacylglycerols (colloquially called ‘fat’
or ‘oil’, depending on their aggregation state) are inert and
sufficiently hydrophobic to provide optimal packaging of
FFAs for efficient transport and storage. Essentially all cells
are able to incorporate fatty acids into triacylglycerols.
In vertebrates, the vast majority (>90%) of ­triacylglycerols
are deposited in white adipose tissue (WAT).
During feeding, WAT stores excessive calories as fat.
Conversely, during fasting, these fat stores are degraded
to supply the body with FFAs. The release of FFAs
from triacylglycerols requires ester hydrolysis by an
enzymatic process called lipolysis. Given that triacyl­
glycerol molecules are themselves unable to cross bio­
logical membranes, their transport in and out of cells
also requires lipolysis and resynthesis. In physiological
terms, lipolysis fulfils three major functions in verte­
brates: gastro­intestinal lipolysis mediates the absorp­
tion of nutritional fats and oils in the stomach and gut 2,
vascular lipolysis enables the cellular uptake of FFAs
and mono­acylglycerols from circulating triacylglycerol-
rich plasma lipoproteins (very-low-density lipoproteins
(VLDLs) and chylomicrons)3, and intracellular l­ipolysis
accounts for the release of FFAs from cytoplasmic lipid
droplets (cLDs) and from endocytosed lipoprotein-­
associated triacylglycerols3. Cytoplasmic lipases hydrolyse
cLD-associated triacylglycerols at pH ~7 in a process
called ‘neutral’ lipolysis 4. By contrast, lipoprotein-­
associated triacylglycerols are transported by early and
late endosomes to the lysosome, where they undergo
‘acid’ lipolysis — hydrolysis by lysosomal acid lipase
(LAL) at pH 4.5–5 (REF. 5). Historically, these distinct
pathways were considered separate and were thought to
have little functional overlap. However, the discovery of
lipophagy in 2009 (REF. 6) suggested that acid lipoly­sis
is not restricted to the catabolism of endocytosed lipo­
proteins. Lipophagy is a subtype of macroautophagy

NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 18 | NOVEMBER 2017 | 671

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
REVIEWS

• Membrane integrity • Growth and

• Storage and detoxification and fluidity development
of FFAs in lipid droplets • Biosynthesis of steroid • Vision
• Emulsification in lipoproteins Membrane-lipid hormones, bile acids, • Glycoprotein Endocannabinoid • Glycolysis
for transport synthesis vitamin D synthesis signalling • Gluconeogenesis
Acid lipolysis Neutral lipolysis

CE, RE Cholesterol, retinol

TAG DAG MAG Glycerol
O
O H2O HO H O HO H O HO
O ATGL O 2
HSL 2
MGL
O O HO HO
O LAL O LAL O
O O O HO
CE, RE
• Lipid signalling, such as via PPARs
• Lipotoxicity
• Activation to acyl-CoA
- mitochondrial transport and β-oxidation
- protein acylation and acetylation FFA
O
- synthesis of membrane lipids,
OH
prostaglandins, leukotrienes, ceramides

Figure 1 | Overview of the enzymes involved in lipolysis and the potential functions of their enzymatic products.
Neutral lipolysis (upper panel): adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and monoacylglycerol
lipase (MGL) consecutively hydrolyse triacylglycerol (TAG), diacylglycerol (DAG) and Nature Reviews | Molecular
monoacylglycerol (MAG),Cell Biology
respectively, to generate non-esterified free fatty acids (FFAs) and glycerol. Acid lipolysis (lower panel): lysosomal acid
lipase (LAL) hydrolyses both TAG and DAG. Whether LAL is also able to hydrolyse MAG is controversial. HSL and LAL
additionally catalyse the hydrolysis of cholesteryl ester (CE) to cholesterol and retinyl ester (RE) to retinol. The potential
utilization of lipolytic intermediates and products is indicated. PPARs, peroxisome proliferator-activated receptors.

that facilitates the transport of cLD components, such a severe lipolytic defect and leads to systemic triacyl­
as triacylglycerols or cLD proteins, to lysosomes for glycerol accumulation in WAT and non-adipose tissues.
White adipose tissue
(WAT). A loose connective ­subsequent degradation by lysosomal enzymes. Triacylglycerol accrual in Atgl−/− animals is most pro­
tissue that predominantly Key discoveries during the last decade have reshaped nounced in cardiac muscle, resulting in mitochondrial
consists of adipocytes our understanding of many aspects of fat catabolism in dysfunction and lethal cardiomyopathy 3 months after
and that stores excess cells. This Review focuses on the principal pathways of birth. Global ATGL deficiency also causes severe cold
nutrients as triacylglycerols.
intracellular lipolysis — neutral lipolysis of cLDs and sensitivity. Whether defective thermoregulation results
Very-low-density acid lipolysis of cLDs (lipophagy) as well as lipo­protein- from brown adipose tissue dysfunction or impaired
lipoproteins associated lipids — and their crosstalk. Also discussed ­cardiac and haemodynamic function is not known.
(VLDL). Liver-derived plasma are new insights that are particularly relevant to both Many proteins regulate neutral lipolysis either directly
lipoproteins of very low density
understanding of the pathogenesis of metabolic dis­ or indirectly (FIG. 2). Two proteins directly interact with
that transport lipids
(predominantly eases and the exploration of new treatment strategies the patatin domain of ATGL and control its enzymatic
triacylglycerols) from the liver for lipid-associated disorders. activity in opposite ways: lipid droplet-binding protein
to non-hepatic tissues. CGI‑58 (also known as ABHD5) activates ATGL12 by an
Neutral lipolysis in the cytoplasm unknown mechanism, whereas G0/G1 switch pro­
Chylomicrons
Intestine-derived plasma
The discovery of adipose triglyceride lipase (ATGL; also tein 2 (G0S2) inhibits the enzyme13 (FIG. 2a). Consistent
lipoproteins that transport known as patatin-like phospholipase domain-­containing with a role for CGI‑58 in ATGL activation, CGI‑58−/−
dietary lipids (predominantly ­protein 2 (PNPLA2))7–9 in 2004, and the informative mice develop a lipid storage phenotype similar to the
triacylglycerols) from the phenotype of ATGL-deficient mice10 established the one observed in Atgl−/− mice, albeit less pronounced14.
digestive tract (intestine)
current concept of lipolysis in adipose and non-adipose In sharp contrast to Atgl−/− mice, however, CGI‑58−/− mice
to the liver and other tissues.
tissues. ATGL initiates triacylglycerol hydrolysis to form additionally exhibit severe ichthyosis, trans-­epidermal
Lipases diacylglycerol and FFAs. Hormone-sensitive lipase water loss and postnatal death around 16 hours after
Enzymes that hydrolyse fatty (HSL) and monoacylglycerol lipase (MGL)11 complete birth. The epidermal skin defects can be restored by
acid–glycerol esters. the process by consecutively hydrolysing diacyl­glycerols the exclusive expression of CGI‑58 in keratinocytes15.
Patatin-like phospholipase
into monoacylglycerols and FFAs and hydrolysing Although the ATGL-independent biochemical func­
domain-containing protein 2 ­monoacylglycerols into glycerol and FFAs (FIG. 1). tion of CGI‑58 in the epidermis remains unknown, it is
(PNPLA2). A protein with a interesting to note that deficiencies in PNPLA1 (REF. 16)
patatin domain that hydrolyses ATGL initiates triacylglycerol hydrolysis. ATGL belongs (another PNPLA family member, of unknown biochem­
neutral lipids, phospholipids
to a family of nine PNPLA genes and is the only robust ical function) and diacylglycerol O-acyltransferase 2
and retinyl esters.
triacylglycerol hydrolase within the family. ATGL has (DGAT2, which is an enzyme essential for the final step
Patatin domain an unusual Ser–Asp catalytic dyad in its patatin domain of triacylglycerol synthesis)17 in mice cause a very similar
A protein domain of instead of the more classical Ser–Asp–His catalytic skin phenotype. These observations suggest that all three
approximately 180 amino triad present in other lipid hydrolases. The pheno­ proteins act within the same, currently undefined bio­
acids in length that was
originally discovered
type of Atgl−/− mice clearly highlighted the crucial role chemical pathway in the epidermis. ATGL and CGI‑58
in the potato tuber storage of this enzyme in mammalian lipolysis and energy deficiencies in humans manifest clinical features similar
protein patatin. homeostasis10. Global ATGL deficiency in mice causes to those observed in the genetic mouse models (BOX 1).

672 | NOVEMBER 2017 | VOLUME 18 www.nature.com/nrm

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
 REVIEWS

Brown adipose tissue
Adipose tissue involved in
thermoregulation, uncoupling
mitochondrial electron
transport from ATP synthesis
and thereby generating heat
during chronic cold exposure.
sn
A notation that stands
for ‘stereospecific numbering’
and describes the
stereochemical configuration
of chiral glycerol derivatives.
Human G0S2 was discovered in expression studies
in blood mononuclear cells during cell cycle re‑entry
(G0‑to‑G1 transition)18. G0S2 suppresses lipolysis by
inhibiting ATGL13 (FIG. 2a). Subsequent studies showed
that a 32‑amino-acid fragment of G0S2 is sufficient to
interact with the patatin domain of ATGL and non-
competitively inhibit its activity 19. CGI‑58 and G0S2
bind to different sites on ATGL20. Overexpression and
knockout studies in mutant mouse models demon­
strated that G0S2 also regulates lipolysis in various
­tissues in vivo21–24.
Other proteins regulate ATGL indirectly. FFA-
binding proteins (FABPs) expressed in the liver, intes­
tine, muscle, adipose tissue and skin (FABPs 1–5)
interact with CGI‑58 (REF. 25) (FIG. 2a). This interaction
activates ATGL by a currently unknown mechanism.
Pigment epithelium-derived factor (PEDF) also activ­
ates ATGL, although the mechanism by which this
occurs is controversial. One study showed that secreted
PEDF binds to ATGL and unleashes the phospho­lipase
activity of the enzyme at the plasma membrane 26,
whereas other studies concluded that the ATGL–PEDF
interaction occurs on cLDs and induces triacylglycerol
hydrolysis27,28. Fat-specific protein FSP27 (also known
as CIDE‑C) inhibits ATGL by a mechanism that is also
controversial. Whereas one study claimed that FSP27
interacts with the transcription factor early growth
response protein 1 (EGR1), leading to the repression of
ATGL expression29, another argued that FSP27 directly
interacts with ATGL to inhibit its hydrolytic activity 30.
HSL and MGL complete the lipolytic enzyme trio.
ATGL possesses narrow substrate specificity for triacyl­
glycerols containing long-chain fatty acids, preferentially
cleaving ester bonds in the sn‑1 or sn‑2 position31. The
enzyme poorly hydrolyses diacylglycerols and mono­
acylglycerols, so other lipases are needed to release the
remaining fatty acids from the glycerol backbone. The
main diacylglycerol hydrolase is HSL (FIG. 2b), which
predominantly cleaves fatty acid residues in the sn‑1 or
sn‑3 position of diacyl­glycerols32. HSL exhibits much
broader substrate specificity than ATGL and can hydro­
lyse ester bonds in triacylglycerols, diacylglycerols,
monoacyl­glycerols, cholesteryl esters, retinyl esters and
short-chain carbonic acid esters33 (FIG. 1). The findings
that HSL cannot compensate for an absence of ATGL
to prevent systemic lipid accumulation and that HSL
deficiency results in diacyl­glycerol, rather than triacyl­
glycerol, accumulation in various tissues of ATGL-
deficient mice and humans10,34 indicated that HSL has
a predominant role in diacylglycerol catabolism within
the lipolytic ­cascade. Lack of hydrolysis of cholesteryl
esters and retinyl esters in the testes of HSL-deficient
mice leads to defective spermatogenesis and infertility 35.

a b c d
Starvation Starvation NPs Starvation

cAMP
TFEB TFEB TFEB
PKA PKG cAMP cGMP
PPARs– PPARs– PPARs– PPARs–
cAMP SIRT1 FABPs PGC1α PGC1α PGC1α SIRT1 PGC1α
P P
PKA PKG ERK FABP4
Perilipin 1 PEDF
Ac Ac
FOXO1 AMPK FOXO1
P Ser659 Ser660
P
CGI-58 P P P P
Ser600
ATGL Ser406 Ser563 HSL MGL LAL
PDE3B G0S2
Ser565 P
cAMP
CAMK2
mTORC1, EGR1 GSK4
mTORC2 OxLDL AggLDL
PDE3B AMPK
FSP27 Lysosomes

Insulin, IGF Nutrient abundance Insulin, IGF Nutrient abundance

Figure 2 | Direct and indirect regulation of the lipases involved in neutral
and acid lipolysis. a | Transcription of adipose triglyceride lipase (ATGL)
is controlled by sirtuin 1 (SIRT1)-mediated deacetylation of forkhead box
protein O1 (FOXO1) and by peroxisome proliferator-activated receptor
(PPAR) and PPARγ co-activator 1α (PGC1α). ATGL enzyme activity is regulated
by direct interaction of ATGL with its co-activator, lipid droplet-binding
protein CGI‑58, and with its inhibitor, G0/G1 switch protein 2 (G0S2), as well
as by phosphorylation of Ser406 by AMP-activated protein kinase (AMPK).
Other factors, including pigment epithelium-derived factor (PEDF),
fat-specific protein FSP27, fatty acid-binding proteins (FABPs), mTOR
complex 1 (mTORC1) and mTORC2 and early growth response protein 1
(EGR1), also regulate ATGL activity. b | Transcription of hormone-sensitive
lipase (HSL) is also regulated by the PPAR–PGC1α axis. In response to
starvation, protein kinase A (PKA) activates HSL by phosphorylation of Ser
Nature Reviews | Molecular Cell Biology
residues 563, 659 and 660. Natriuretic peptides (NPs) stimulate lipolysis
through PKG and phosphorylation of HSL at Ser660. AMPK, glycogen
synthase kinase 4 (GSK4) and calcium/calmodulin-­dependent protein kinase
type II (CAMK2) inhibit HSL by phosphorylating Ser565. FABP4 interacts
with HSL and stimulates its activity. c | Monoacylglycerol lipase (MGL)
expression is responsive to PPAR–PGC1α activation. d | Similar to
ATGL expression, lysosomal acid lipase (LAL) expression is also positively
regulated by transcription factor EB (TFEB), PGC1α and SIRT1‑dependent
deacetylation of FOXO1. Oxidized (Ox) or aggregated (Agg) low-density
lipoprotein (LDL) inhibits LAL activity by changing the lysosomal pH.
Ac, acetylation; cAMP, cyclic adenosine monophosphate; GF, insulin-like
growth factor; P, phosphorylation; PDE3B, phosphodiesterase 3B.

NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 18 | NOVEMBER 2017 | 673

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
REVIEWS

Endocannabinoid signalling
Signalling that comprises
cannabinoid receptors and
their endogenous ligands,
which regulate numerous
physiological processes,
including energy homeostasis,
pain perception, inflammation
and tumorigenesis.
Catecholamines
Tyrosine derivatives of
catechol, including adrenaline,
noradrenaline and dopamine,
that act as neuromodulators
and hormones.
Natriuretic peptides
Peptides that control the
homeostasis of water, sodium
and potassium in the body.
In adipocytes, these peptides
also regulate lipolysis.
Gene mutations leading to HSL deficiency in humans
were recently reported36 (BOX 1). Even though HSL-
deficient mice and humans exhibit similar phenotypes,
unlike mice, HSL-deficient human males are fertile36.
The final step in the lipolytic cascade is catalysed by
MGL, which cleaves both triacylglycerol-derived and
glycerophospholipid-derived monoacylglycerols in all
three stereospecific positions11. Owing to the increased
solubility of monoacylglycerols compared with triacyl­
glycerols and diacylglycerols, the enzyme is cyto­plasmic
and, unlike ATGL or HSL, does not act on cLDs. MGL
fulfils a crucial role in endocannabinoid signalling by
hydrolysing and inactivating 2­ ‑arachidonylglycerol,
which is the most abundant endocannabinoid in
mammals, regulating a wide variety of biological pro­
cesses, including energy homeostasis, pain perception,
­inflammation and tumorigenesis37.
Direct and indirect regulation of neutral lipolysis.
Endocrine regulation of neutral lipolysis is complex
and involves numerous hormones, growth factors
and (adipo)cytokines that are linked to diverse signal
transduction pathways33 (FIG. 2). The majority of stud­
ies addressed the control of lipolysis by catecholamines
and insulin in WAT during fasting and feeding. In the
fasted state, catecholamines (in WAT, muscle and the
liver) and glucagon (in the liver) increase cellular cAMP
concentrations by activating a Gs protein-dependent
adenylyl cyclase. Next, cAMP activates protein kinase
A (PKA), which phosphorylates a number of cLD-­
associated proteins, including perilipin 1, HSL and
CGI‑58. Unphosphorylated perilipin 1 binds CGI‑58
(REF. 38) and thereby prevents CGI‑58‑mediated ATGL
activation. Perilipin 1 phosphorylation at multiple
residues leads to the release of CGI‑58, which is then
able to activate ATGL39. Simultaneously, phosphoryl­
ated HSL translocates from the cytosol to cLDs. HSL
regulation by enzyme phosphorylation is complex 40
(FIG.  2b) . Five distinct Ser residues are phosphoryl­
ated by either activating kinases (PKA, PKG and
extracellular-­signal-regulated kinases (ERKs)) or inhib­
itory kinases (AMP-activated protein kinase (AMPK),
calcium/calmodulin-­dependent protein kinase type II
and glycogen synthase kinase 4), which respectively
trigger and prevent HSL translocation and activation
(FIG. 2b). In addition to catecholamines and glucagon,
other effectors and metabolites are also able to activ­
ate neutral lipolysis via the cAMP–PKA pathway,
including thyroid-stimulating hormone and melano­
cortins33. Natriuretic peptides induce lipolysis through

Box 1 | Pathophysiology of neutral lipolysis

Rare mutations in the genes encoding adipose triglyceride lipase (ATGL), lipid droplet-binding protein CGI‑58 and
hormone-sensitive lipase (HSL) cause distinct metabolic disorders in humans.
• ATGL. Individuals with mutations in the gene encoding ATGL develop neutral lipid storage disease with myopathy
(NLSDM)34. Genetically confirmed NLSDM has been diagnosed in over 40 individuals with approximately 30 different
mutations. NLSDM is a rare autosomal disorder characterized by systemic triacylglycerol accumulation in multiple tissues,
including cardiac and skeletal muscles; the liver, skin and pancreas; and blood leukocytes (Jordan anomaly)164. Cardiac
steatosis is associated with severe dilated cardiomyopathy in 44% of patients, and this condition often requires heart
transplantation to avert cardiac death. Progressive skeletal myopathy is observed in the majority of patients and often
leads to loss of muscle strength and difficulties in walking. Other more moderate and rare clinical features include
hepatosteatosis and hepatomegaly, pancreatitis and cognitive impairment. Unlike Atgl−/− mice, ATGL-deficient humans
are mostly normoglycaemic and show no signs of increased insulin sensitivity or glucose tolerance165.
• CGI‑58. Human CGI‑58 deficiency causes neutral lipid storage disease with ichthyosis (NLSDI; also designated
Dorfman-Chanarin syndrome). All individuals with NLSDI exhibit severe congenital ichthyosiform erythroderma166,167.
More infrequent clinical features include hepatosplenomegaly, myopathy, hearing loss and mental retardation.
Although patients with NLSDI also exhibit systemic triacylglycerol accumulation and Jordan anomaly, lipid
accumulation in muscle and the heart does not lead to severe skeletal or cardiac myopathy. In 2001, mutations in
CGI‑58 were found to cause NLSDI168. Unlike NLSDM, NLSDI always occurs in conjunction with a severe skin phenotype,
supporting an ATGL-independent function of CGI‑58 in the epidermis. Similar to CGI‑58−/− mice14, the epidermal
keratinocytes of humans with NLSDI are unable to synthesize acylceramides or to form a functional corneocyte lipid
envelope169. Although the pathophysiological consequences of CGI‑58 deficiency are well established, the actual
biochemical function of CGI‑58 remains a subject of debate. The postulated ATGL-independent functions of CGI‑58
include enzyme activity as an acyltransferase and/or involvement in autophagy145,170,171. To date, however, a link between
these activities and the skin pathology observed in patients with NLSDI remains elusive.
• HSL. Mutations causing HSL deficiency were first reported only recently36,172. Individuals with homozygous mutations
in HSL exhibit a relatively benign phenotype. Similar to Hsl−/− and Atgl−/− mice, HSL-deficient patients develop partial
lipodystrophy owing to a reduction in peroxisome proliferator-activated receptor γ (PPARγ)-driven triacylglycerol
synthesis in HSL-deficient adipocytes, reinforcing the links among lipolysis, PPAR signalling and lipid synthesis. Unlike
Hsl−/− mice, however, all HSL-deficient humans develop insulin resistance and type 2 diabetes, fatty liver disease and
dyslipidaemia, characterized by increased plasma triacylglycerol and low-density lipoprotein concentrations and
decreased plasma high-density lipoprotein concentrations. It is conceivable that this metabolic syndrome phenotype is
a consequence of the partial lipodystrophy affecting HSL-deficient individuals. A similar phenotype in knockout mice
may have been missed because plasma lipid and lipoprotein parameters were studied in young mice, before the onset
of lipodystrophy. It is also interesting to note that HSL-deficient human males are fertile, whereas male Hsl−/− mice are
sterile35. Functional spermatogenesis and fertility in mice require the presence of enzymatically active HSL in the testes,
which is apparently not the case in human males with HSL deficiency173.

674 | NOVEMBER 2017 | VOLUME 18 www.nature.com/nrm

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
 REVIEWS

the synthesis of cyclic guanosine monophosphate
and the activation of PKG, which phosphorylates peri­
lipin 1 and HSL41 (FIG. 2b). Finally, pituitary growth hor­
mone (somato­tropin) activates ATGL by inducing its
gene transcription via signal transducer and activator
of ­transcription 5 (STAT5) signalling 42.
Inhibitors of lipolysis include the classical ­hormone
inhibitors insulin and insulin-like growth factors43
as well as non-hormone inhibitors like lactate, adeno­
sine, β‑hydroxybutyrate and nicotinic acid (niacin),
which act through G-protein-coupled receptors 33.
Although known for decades, the complexity of insulin-­
mediated inhibition of lipolysis remains incompletely
understood. According to the established model,
insulin induces PKB-dependent phosphorylation and
activation of phosphodiesterase 3B (PDE3B), which
subsequently hydrolyses cAMP (FIG. 2a,b). This event in
turn leads to decreased phosphorylation of perilipin 1
and HSL by the inhibition of PKA as well as the induc­
tion of protein phosphatase 1 and downregulation of
lipolysis. Consistent with this model, insulin-mediated
inhibition of lipolysis was recently shown to require
the activation of PDE3B44. However, PDE3B activation
did not depend on PDE3B phosphorylation by PKB44.
Accordingly, the mech­anism of PDE3B activation by
insulin remains unknown. Insulin additionally inhibits
ATGL mRNA synthesis in WAT and the liver by exerting
an inhibitory effect on the transcription factor forkhead
box protein O1 (FOXO1), which is a direct activator of
ATGL transcription45 (FIG. 2). Conversely, FOXO1 activ­
ation by deacetylation mediated by NAD+-dependent
protein deacetylase sirtuin 1 (SIRT1) induces ATGL
expression46, thereby providing an explanation for the
pro-lipolytic effect of SIRT1 (REF. 47) and the SIRT1
­activator resveratrol48.
Other major insulin-stimulated kinases that regulate
neutral lipolysis include mTOR complex 1 (mTORC1),
mTORC2 (REF. 49) and AMPK50 (FIG. 2). The inhibition
of mTORC1 by rapamycin or inactivation by raptor
deletion increase neutral lipolysis and FFA release from
adipocytes51; conversely, mTORC1 activation through
overexpression of GTP-binding protein Rheb decreases
lipolysis, supporting an antilipolytic role for mTOR
signalling 51. The signalling mechanism is complex and
incompletely understood but involves transcriptional
regulation of ATGL by EGR152. Similarly, mTORC2
inactivation in WAT due to adipose-specific loss of the
mTORC2 subunit rictor also resulted in unrestrained
lipolysis, supporting an important antilipolytic role for
mTOR signalling during nutrient abundance53.
The role of AMPK in the regulation of lipolysis is
less well defined. AMPK is activated during fasting and
exercise, when cellular AMP concentrations increase,
but whether or not this induction contributes to the
upregulation of lipolysis is controversial. Contradictory
reports claim that the kinase induced, inhibited or
did not affect neutral lipolysis in WAT50. The findings
that the lipolytic response depends on the duration of
experi­mental AMPK inhibition or activation54, that
AMPK stimulates ATGL but inhibits HSL55 (FIG. 2a,b) and
that the HSL phosphorylation pattern differs between
adipose tissue and muscle56 may at least partially under­
lie the confusion. It is also uncertain whether AMPK
or PKA ­phosphorylates ATGL at Ser406 to regulate
its activity 57,58.
Whereas ATGL and HSL are regulated by multi­
ple mechanisms, MGL regulation is less complex
(FIG. 2c). Tissue-specific isoforms of the enzyme exist 59,
but whether this variation affects activity is not clear.
Posttranslational modifications have not been linked to
MGL function. Transcriptionally, MGL is a target for
the nuclear receptor peroxisome proliferator-activated
receptor α (PPARα)60.
Neutral lipolysis regulates key signalling pathways in
energy homeostasis. In addition to having a ­crucial role
as energy substrates, the products and inter­mediates
of neutral lipolysis impact lipid signalling and energy
homeo­stasis. For example, a functional link exists
between lipolysis, PPAR signalling and oxidative ­capacity
in various tissues. Defective lipolysis in the heart of Atgl−/−
(REF. 61) and CGI‑58−/− (REF. 62) mice not only causes
severe fat accumulation but also leads to impaired mito­
chondrial respiratory function. This insufficiency results
from pronounced downregulation of PPARα ­target
genes involved in mitochondrial substrate oxid­ation
and oxid­ative phosphorylation. PPARα agonists such as
WY14643 and fenofibrate reverse the phenotype, con­
sistent with the concept that mitochondrial dysfunction
contributes to the lethal cardiomyopathy in Atgl−/− mice.
This conclusion is further supported by the observation
that cardiomyocyte-exclusive expression of ATGL pre­
vents severe cardiomyopathy and premature death63.
Decreased expression of PPARα target genes was also
observed in the liver 64,65, intestinal cells66, macrophages67
and brown adipocytes57,68 of Atgl−/− mice. Hepatic PPARα
signalling not only is affected by tissue-autonomous
lipolysis in the liver but also depends on lipolysis in
and FFA delivery from WAT. Conditional knockout of
the genes encoding ATGL69 or CGI‑58 (REF. 70) in WAT
drastically lowers hepatic PPARα activity and target
gene expression. In pancreatic β-cells, ATGL regulates
mitochondrial function and glucose-­stimulated insulin
secretion through PPARδ71, and β-cell-specific deletion
of ATGL causes hyperglycaemia, which is reversible by
the ­administration of PPARδ (but not PPARα) agonists.
ATGL-mediated lipolysis also affects PPARγ sig­
nalling and lipid synthesis in WAT. Mice with global or
adipose tissue-specific ATGL deficiency are resistant to
obesity induced by a high-fat diet owing to decreased
food intake and downregulation of FFA uptake and
triacyl­g lycerol synthesis in WAT69,72. The defect in
PPARγ signalling and lipid synthesis in WAT can be
abrogated by the PPARγ agonist rosiglitazone72. This
finding is reminiscent of the impaired PPARγ signal­
ling and lipodystrophy that occurs in HSL-deficient
mice and humans36,73 and supports a more general func­
tion for lipolysis in PPAR signalling, beyond the activity
of ­individual lipases.
The signal transduction pathways linking lipolysis
to PPAR activity remain unclear. The evidence that
FABPs bind to CGI‑58, thereby affecting PPARα and

NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 18 | NOVEMBER 2017 | 675

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
REVIEWS

Starvation acetyl-proteome82–84. Histone acetylation at specific Lys

residues is a highly regulated process that requires, in
addition to acetyl-CoA, the activity of more than a
P
AMPK ULK1
Autophagy, dozen histone acetyltransferases. The abundance of
mTOR lipophagy cyto­plasmic acetyl-CoA affects histone acetylation and
P regu­lates the transcription of numerous genes85,86, but
TFEB P SIRT1
Ac the specific effect of lipolysis on histone acetylation
PGC1α
­patterns remains to be elucidated.
P Ac Unlike histone acetylation, protein acetylation out­
TFEB PGC1α PPARα FOXO1 side the nucleus occurs at low stoichiometry, likely by a
non-enzymatic mechanism that is mostly driven by
PGC1α local acetyl-CoA concentrations82,83. Consistent with
this concept, non-nuclear protein acetylation is highest
PPARα
RXR

in mitochondria and is markedly affected by fasting-­

TFEB FOXO1
ATGL LAL
HSL
MGL
Neutral Acid
lipolysis lipolysis
Figure 3 | Common regulation of neutral lipolysis and lipophagy. During nutrient
Nature Reviews | Molecular Cell Biology
starvation, inhibition of the mTOR signalling pathway allows the dephosphorylation
and nuclear translocation of transcription factor EB (TFEB). Starvation also activates
AMP-activated protein kinase (AMPK), which phosphorylates peroxisome proliferator-
activated receptor γ (PPARγ) co-activator 1α (PGC1α), thereby activating gene
transcription mediated by PPARα–PGC1α–retinoid X receptor (RXR). AMPK enhances
sirtuin 1 (SIRT1) activity, leading to the deacetylation of downstream SIRT1 targets,
including PGC1α and the forkhead box O (FOXO) transcription factors. After
translocation to the nucleus, TFEB, PPARα–PGC1α–RXR and the FOXO transcription
factors initiate the transcription of adipose triglyceride lipase (ATGL) and lysosomal
acid lipase (LAL), resulting in increased neutral lipolysis and acid lipolysis, respectively.
AMPK induces lipophagy by phosphorylating and activating serine/threonine-protein
kinase ULK1.
PPARγ signalling in hepatocytes and fat cells25, supports
lipolysis-­coupled transport of FFAs into the cell nucleus,
where they serve as activating ligands for PPARs.
Consistent findings show that PPAR activity requires the
nuclear translocation of FFA-loaded FABPs74–76. FABP4
also interacts with HSL and stimulates its enzyme activ­
ity, but it is not known whether this interaction regulates
PPAR activity77,78. Additionally, lipolysis activates SIRT1
(REF. 79), which deacetylates and thereby activates PPARγ
co-activator 1α (PGC1α). SIRT proteins also induce the
activity of FOXO transcription factors, which regu­
late multiple processes in adipogenesis and adipose
metabolism, including autophagy 80 and the expression
of ATGL46 and LAL81. Accordingly, ATGL, sirtuins and
FOXO1 form an autoregulatory loop that coordinates
lipid metabolism and links neutral lipolysis and acid
lipolysis (FIG. 3).
Protein acetylation. Lipolysis may also regulate meta­
bolic pathways via protein acetylation. The oxidation
of lipolysis-derived FFAs generates acetyl-CoA, which
is the sole cofactor for protein acetylation. It is there­
fore not surprising that lipolysis affects the cellular
induced lipolysis and FFA oxidation. Fasting increases
mitochondrial protein acetylation, but this effect is
completely attenuated in Atgl−/− mice84 and in mice that
lack very-long-chain acyl-CoA dehydrogenase, which
is a key enzyme for the initiation of FFA ­β-oxidation87.
This finding supports the view that lipolysis and
acetyl-CoA production drive mitochondrial protein
acetylation. During fasting or calorie restriction, when
mitochondrial acetyl-CoA concentrations are high,
­target sites in multiple proteins are hyperacetylated and
require subsequent deacetylation (‘repair’) by the mito­
chondrial deacetylase SIRT3 (REFS 84,88). Consistent
with the increased demand for such repair, SIRT3 is
upregulated during fasting and calorie restriction88,89.
Increased hepatic acetyl-CoA concentrations owing to
increased adipose lipolysis adversely affect insulin con­
trol of hepatic glucose production90. Whether this effect
involves differences in protein acetylation in response to
acetyl-CoA concentrations is not known.
Acid lipolysis in lysosomes
In addition to cLDs, endosomes and lysosomes are the
only cell organelles where intracellular triacyl­glycerol
hydrolysis occurs. Neutral lipolysis was thought to be
restricted to the catabolism of cLD-associated triacyl­
glycerols, whereas lysosomal (acid) lipolysis was
assumed to be exclusively responsible for the degrad­
ation of exogenous plasma lipoprotein-associated lipids,
including triacylglycerols. The discovery of lipophagy 6
challenged this view and demonstrated that lyso­somal
triacylglycerol degradation also contributes to the
­turnover of c­ ytosolic lipid stores.
Lysosomal triacylglycerol degradation is carried out
by LAL owing to its optimal activity at the lysosomal pH
of 4.5–5. LAL is highly glycosylated and exists in vari­
ous tissue-specific isoforms. It exhibits broad substrate
specificity, hydrolysing triacylglycerols, diacyl­glycerols,
cholesteryl esters91,92 and retinyl esters93 (FIG. 1). Whether
LAL also hydrolyses monoacylglycerols is contro­
versial91,92. LAL localization is not restricted to lyso­
somes, as it can be secreted from cells via the classical
endo­plasmic reticulum (ER)–Golgi secretory pathway
and can subsequently re-enter cells and lysosomes by
endocytosis94. Furthermore, LAL can be secreted from
cells by a process called exophagy, in which lysosomes
fuse with the plasma membrane and release their content
into the extracellular space95.

676 | NOVEMBER 2017 | VOLUME 18 www.nature.com/nrm

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
 REVIEWS

Scavenger receptors Exogenous triacylglycerols, cholesteryl esters and
Lipoprotein receptors that r­ etinyl esters destined for LAL-mediated degradation
remove modified lipoproteins enter cells by receptor-dependent uptake of plasma lipo­
(for example, acetylated proteins. Lipoprotein binding by various receptors, such
or oxidized low-density
lipoproteins) and other
as the low-density lipoprotein (LDL) receptor, LDL
negatively charged receptor-related proteins or scavenger receptors; subse­
macromolecules from quent lipoprotein internalization; and the recycling of
the blood. endosomal receptors have been extensively reviewed else­
where96,97. Although the intracellular sorting mechanisms
Kupffer cells
Specialized macrophages
for all lipoprotein receptors are not equally well studied,
in the liver. it is generally understood that when late endosomes even­
tually fuse with lysosomes, lysosomal hydrolases degrade
Foam cell all lipoprotein components, including apolipoproteins,
A lipid-filled macrophage that
is present in atherosclerotic
phospholipids and neutral lipids. To date, the only known
lesions and plaques. enzyme involved in the hydrolysis of neutral lipids is LAL.
LAL regulation. An absence or dysfunction of LAL
results in excessive lysosomal lipid accumulation in
humans and mice, consistent with the notion that LAL
is the sole enzyme responsible for degrading triacyl­
glycerols and cholesteryl esters in lysosomes. LAL defi­
ciency in humans causes Wolman disease and cholesteryl
ester storage disease (CESD) (BOX 2). Lal−/− mice survive
Box 2 | Pathophysiology of acid lipolysis
Mutations in the lysosomal acid lipase (LAL)-encoding gene (LIPA) cause two diseases in
humans that differ in severity. Wolman disease is a lipid storage disorder174 characterized
by a failure to thrive, progressive hepatosplenomegaly, adrenal calcification and death
within the first year after birth. Cholesterol ester storage disease (CESD) has a similar
phenotype but a later onset and a much slower progression175. Both conditions are
caused by reduced LAL activity, giving rise to excessive cholesteryl ester and
triacylglycerol accumulation in lysosomes176,177. Whereas individuals with Wolman
disease invariably lack the enzyme and require liver transplantation to survive,
individuals with CESD frequently, but not always, exhibit some remnant enzyme
activity178. The spectrum of disease manifestation and progression in CESD ranges from
severe liver disease and liver failure in children to no apparent clinical features until
adulthood. The outcome is often independent of the magnitude of remnant LAL activity.
Therefore, the more recently used designation ‘LAL deficiency’ (LAL‑D) seems more
appropriate for both conditions. Unlike Lal−/− mice, which develop severe steatohepatitis,
characterized by massive macrophage infiltration and inflammation, humans with LAL‑D
develop liver steatosis and fibrosis but show no signs of liver inflammation178,179.
Individuals with CESD develop dyslipidaemia characterized by increased plasma
low-density lipoprotein (LDL) cholesterol concentrations and decreased plasma
high-density lipoprotein (HDL) cholesterol concentrations, resulting in increased
atherosclerosis178. LAL has a crucial role in cholesterol homeostasis by linking
exogenously internalized cholesterol and cholesteryl esters to the regulation of de novo
cholesterol synthesis pathways (see main text). Excessive uptake of modified LDL
in macrophages via the scavenger receptor pathway has been shown to inhibit LAL in
lysosomes and extracellular compartments. In macrophages, this derepresses de novo
cholesterol synthesis, increases LDL receptor activity and attenuates HDL-mediated
reverse cholesterol transport. Together, these events lead to increased foam cell
and coronary lesion formation and an increased risk of coronary artery disease.
Enzyme replacement therapy was recently approved for the treatment of individuals
with LAL-D180. Similar treatment strategies have been previously approved for other
lysosomal storage diseases, such as Fabry disease and Pompe disease. In a
placebo-controlled trial, 66 patients were treated with an enzymatically active LAL
preparation (sebelipase)180. Although the mechanism of cellular enzyme uptake
remains unknown, sebelipase treatment was beneficial for many patients with LAL-D.
It improved liver function, normalized plasma transaminase activities and decreased
hepatic triglyceride content. Whether this (very costly) treatment will affect the
long-term outcome or end points of the disease in more severely affected individuals
requires further investigation. Sebelipase may also be a treatment strategy for
atherosclerosis, which is commonly observed in individuals with LAL-D179.
the postnatal state, and the lipid accumulation pheno­
type develops more gradually in these mice than in
humans98,99. Notably, Kupffer cells, and not hepatocytes,
are the main liver cell type in which cholesteryl esters
and triacylglycerols accumulate, which raises both the
question of how LAL-deficient hepatocytes manage
to cope with constant encounters with lipoprotein-­
associated lipids and the possibility that LAL is not the
sole enzyme responsible for degrading triacylglycerols
and choles­teryl esters in lysosomes. Additionally, Lal−/−
mice become lipodystrophic99,100.
Lysosomes are unable to store any degradation prod­
ucts, so the catabolic machinery, including LAL, and
lysosomal export mechanisms are constitutively active.
Therefore, the regulation of acid lipolysis, and specifically
LAL, is less complex than the regulation of neutral lipolysis
(FIG. 2). LAL regulation occurs predominantly at the gene
transcription stage. Similar to the regulation of neutral
lipases, fasting promotes FOXO1‑dependent LAL gene
transcription following the SIRT1‑mediated deacetylation
and nuclear translocation of FOXO1 (REF. 81). LAL gene
transcription is also activated by transcription factor EB
(TFEB)101. Whether transcription factor E3 (TFE3) also
regulates LAL gene transcription has not been directly
tested but seems likely considering the highly redun­
dant activities of TFEB and TFE3 (REFS 102,103) and the
fact that in Caenorhabditis elegans, the TFE3 orthologue
HLH‑30 activates lysosomal lipolysis104. When nutrients
are abundant, the mTOR complex is active and phos­
phorylates TFEB and TFE3, which remain cytosolic and
inactive under this condition. Upon fasting, the proteins
are dephosphorylated, translocate to the nucleus and
regulate the transcription of numerous genes (FIG. 3).
In oxidative tissues such as the liver and muscle, TFEB
regulates the expression of LAL and induces the tran­
scription of other transcription factors that are involved
in catabolic pathways, such as PPARα and its c­ o-activator,
PGC1α101,105, which further augment the expression of
LAL and the neutral lipases ATGL, HSL and MGL60.
Few factors are known to regulate LAL post-­
transcriptionally, but oxidized and aggregated LDLs are
important inhibitors (FIG. 2d). Although the details of
related mechanisms are unknown, modified lipoproteins
appear to cause an increase in luminal pH in lysosomes
owing to decreased vacuolar H+-ATPase-mediated pro­
ton pumping and lysosomal-to-­membrane leakiness106,107,
leading to decreased LAL activity. The modified LDL-
induced hydrolytic defect results in cholesteryl ester
retention in lysosomes108, induction of de novo choles­
terol synthesis and inhibition of reverse cholesterol trans­
port 109. Taken together, these processes may contribute
to foam cell formation and increased a­ therosclerosis in
individuals with CESD (BOX 2).
LAL-generated lipids regulate cellular lipid homeostasis.
The products of LAL are FFAs, unesterified cholesterol
and retinol (FIG. 1). The lysosomal transport and exit strat­
egies for these highly hydrophobic compounds are only
partially understood. The best-studied lysosomal lipid
export process is the transport and secretion of unesteri­
fied cholesterol by Niemann–Pick disease type C (NPC)

NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 18 | NOVEMBER 2017 | 677

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
REVIEWS

Steatosis
A process describing the
abnormal retention of neutral
lipids (triacylglycerols and
cholesteryl esters) within cells
and tissues.
proteins, which are involved in lysosomal cholesterol
traffick­ing 110. According to the current model, NPC2
binds unesterified cholesterol and shuttles it to the lyso­
somal membrane, where NPC1 facilitates transmembrane
passage. Deficiencies in either NPC1 or NPC2 cause NPC,
which is a severe lysosomal cholesterol storage disorder 110.
In addition to binding NPC proteins, cholesterol binds to
lysosome-associated membrane glycoprotein 1 (LAMP1)
and LAMP2, and similarly to what has been observed
for NPC, the absence of these proteins results in the
accumu­lation of unesterified cholesterol in lysosomes111.
How other LAL products are exported remains largely
unknown. For example, it is unclear whether lysosomal
monoacylglycerol hydrolysis is needed or whether mono­
acylglycerols per se exit lysosomes for further hydrolysis by
MGL in the cytoplasm. Similarly, the mechanisms of the
transport and release of FFAs and retinol remain elusive.
Accumulation of FFAs in NPC1‑deficient late endosomes
originally indicated a role for NPC1 in endosomal FFA
release112. However, normal FFA flux in the fibroblasts
of individuals lacking NPC1 did not support a role for
NPC1 in FFA translocation from lysosomes113, leaving the
­mechanisms of FFA export essentially uncharacterized.
After their release from lysosomes, LAL-generated
lipids regulate numerous metabolic processes and serve
as crucial precursors for the de novo synthesis of lipid
species (FIG.  1). Unesterified cholesterol becomes an
important membrane constituent or, upon membrane
saturation, is re-acylated and deposited as cholesteryl
esters in cLDs. The cholesterol content of the ER in turn
regulates the processing, localization and activity of sterol
regulatory element-binding protein 2, which is an impor­
tant transcriptional activator of cholesterol biosynthesis
enzymes and the LDL receptor 96. Furthermore, the oxid­
ation products of unesterified cholesterol (for example,
27‑OH‑cholesterol) are ligands of nuclear receptors
in the liver X receptor family 114. These receptors affect
­cholesterol homeostasis by inducing the conversion
of cholesterol into bile acids in the liver, upregulating
cholesterol efflux from peripheral cells and increasing
­intestinal ­cholesterol excretion.
Similar to FFAs produced by neutral lipolysis, LAL-
generated FFAs are able to regulate PPARα and PPARα
target gene expression100. However, the consequences of
decreased PPARα signalling in LAL deficiency may be
less severe than in humans or mice with ATGL deficiency
and do not lead to the development of cardiomyopathies.
Notably, a study in C. elegans demonstrated that LAL
is involved in the generation of a specific lipid ligand
(oleoylethanolamine) for nuclear receptors that regu­
late the expression of metabolic genes and longevity 115.
Whether LAL-generated retinol and monoacylglycerols
contribute to nuclear receptor and endocannabinoid
­signalling, respectively, is not known.
Lipophagy
In addition to the ubiquitin–proteasome protein
degrad­ation system, autophagy, which is less substrate
selective, represents the most important catabolic path­
way for vari­ous cellular components. Defective auto­
phagy is associated with numerous diseases, including
neurological disorders, cancer, cardiomyopathies and
metabolic dis­orders116. The major variants of autophagy
include macro­autophagy, chaperone-mediated auto­
phagy (CMA) and microautophagy 117. One form of
macroautophagy, designated ‘lipophagy’, was discovered
in 2009 and was shown to contribute to the hydrolysis of
­triacylglycerols stored in cLDs6.
Lipophagy relies on the same general mechanisms
as macroautophagy, which involves more than 30 ATG-
encoding genes118 (FIG. 4a). Macroautophagy in general
and lipophagy in particular are strongly induced by the
major metabolic kinases mTORC1 and AMPK during
lengthy fasting 119. The activity of these kinases depends
on growth factor signalling, the cellular energy status
(ATP:AMP ratio) and nutrient availability 120. Nutrient-
mediated transcriptional regulation of hepatic autophagy
also occurs through the nuclear receptors PPARα and the
liver X receptors121.
The large size of cLDs impedes their recruitment into
lipoautophagosomes6; therefore, lipophagy recruits only
parts of cLDs (FIG. 4b). The potential mechanisms of and
the recruitment factors involved in cLD fragmentation as
well as the specific cargo targets on cLDs remain unidenti­
fied. Lipoautophagosomes eventually fuse with late
endosomes or lysosomes to form auto­lysosomes (FIG. 4b).
The fusion mechanism is insufficiently understood but
may involve soluble N‑ethylmaleimide-sensitive factor
attachment protein receptors, microtubule-associated
protein light chain 3 (LC3), LAMP1, LAMP2B and
LAMP2C as well as small GTPases, such as Ras-related
protein Rab7a122–125. Within autolysosomes, LAL presum­
ably hydrolyses triacylglycerols and cholesteryl esters,
whereas acid phospholipases and peptidases degrade
cLD-­associated phospholipids and proteins, respectively.
A recent study presented an alternative mech­anism,
in which late endosomes and lysosomes directly bind to
‘primed’ cLDs in a process that requires Rab7a (REF. 123)
(FIG. 4b). This interaction is a transient ‘kiss and run’-like
process that resembles the microautophagy that occurs
in yeast, in which catabolic organelles directly bind to
target structures and hydrolyse them after their incorpor­
ation126. How this process would allow for the inter­action
of LAL with triacylglycerols within the core of cLDs
remains to be elucidated.
A substantial contribution of lipophagy to the catabo­
lism of cellular triacylglycerols and cholesteryl esters
has been demonstrated in various cell types, including
hepato­cytes, enterocytes, macrophages, brown adipo­
cytes and neurons127. Autophagy inhibitors, knockdown
of ATG5 in liver cells and the phenotype of mice with
liver-­specific ATG7 deficiency substantiated the crucial
role of lipophagy in the breakdown of hepatic triacyl­
glycerols6. Inhibition of neutral lipases or autophagy
showed that both processes are activated in hepatocytes
during fasting 6. The quantitative contribution of each
of the lipolytic pathways (cytosolic lipolysis and lipo­
phagy) to overall lipid catabolism is unknown and may
vary consider­ably between different cell types, such as
hepatocytes, macrophages and adipocytes. Generally,
the magnitude of s­ teatosis in mice with knockout of
autophagy-­specific genes is variable and more moderate

678 | NOVEMBER 2017 | VOLUME 18 www.nature.com/nrm

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
 REVIEWS

a Autophagy pro-LC3-I
ATG4
• Insulin
• Growth factors LC3-I
• Amino acids
ATG12 ATG7
ATG7 ATG3

ATG10 LC3-II

mTORC1 ATG5
ATG16 p62
ATG5 Cargo
AMPK p62
ATG16 ATG12

ULK1/2-ATG13–FIP200 WIPIs
LC3 lipidation and
Cytoplasm complex PI3P PE autophagosome formation
Beclin 1 VPS34
ER Phagophore
initiation and elongation

b Lipophagy LAL
Lysosome

cLD Rab7 p62

cLD-cargo p62
LC3-II
Cytoplasm
Lipoautophagosome Autolysosome
ER Phagophore

Lipophagy–lipolysis Perilipin 3
crosstsalk” Perilipin 2 HSC70
Perilipin 2,3
LAMP2A
cLD FFA
LC3-II Acid lipolysis
ATGL– CGI-58
LDL

Receptor-mediated endocytosis
Figure 4 | Autophagy, lipophagy, lipolysis and their crosstalk. a | Autophagy. Nutrient uptake and signalling through
insulin, growth factors and amino acids activate mTOR complex 1 (mTORC1), which inhibits autophagosome assembly by
phosphorylating and inactivating a complex composed of serine/threonine-protein kinase ULK1 or ULK2, ATG13 and FAK
family-interacting protein of 200 kDa (FIP200; also known as RB1CC1). Upon nutrient deprivation, in the absence of growth
factors and in conditions of increasing energy demand, AMP-activated protein kinase (AMPK) becomes active, inhibits
Nature Reviews | Molecular Cell Biology
mTORC1 and activates ULK1. Activated ULK1 phosphorylates ATG13 and FIP200 in ULK1–ATG13–FIP200 and ULK2–
ATG13–FIP200 complexes, which then activate beclin 1. Activated beclin 1 interacts with phosphatidylinositol 3‑kinase
catalytic subunit type 3 (PIK3C3; also known as VPS34). Beclin 1–VPS34 heterodimers generate PI‑3,4,5‑phosphate (PI3P),
which nucleates the formation of autophagosomal membranes. Subsequent autophagosome elongation facilitated by
the binding of PI3P to WD repeat domain phosphoinositide-interacting proteins (WIPIs) and autophagosome maturation
involves the successive activity of two ubiquitin-like conjugation systems. The conjugation of ATG12 to ATG5 by the ligases
ATG7 and ATG10 and subsequent binding of the complex to the multimeric protein ATG16 and conjugation of
proteolytically processed microtubule-associated protein light chain 3 (pro‑LC3‑I) to phosphatidylethanolamine (PE)
by the ligases ATG7 and ATG3 lead to LC3‑II formation. The incorporation of LC3‑II into the growing membrane and the
recruitment of cargo adaptor proteins such as p62 lead to the enclosure of cargo (cytoplasmic components or cell
organelles) within autophagosomes. b | Lipophagy involves LC3‑II‑positive membranes engulfing small cytosolic lipid
droplets (cLDs) or sequestering portions of large cLDs. Lipoautophagosomes deliver cLD cargo to lysosomes, wherein
lysosomal acid lipase (LAL) degrades the lipid cargo. Subsequently, non-esterified free fatty acids (FFAs) are released into
the cytosol. Alternatively, lysosomes directly bind to ‘primed’ cLDs via a ‘kiss and run’ mechanism in a process that requires
Ras-related protein Rab7a. Crosstalk between lipophagy and neutral lipolysis is established when LC3‑II‑positive
membranes direct adipose triglyceride lipase (ATGL) to cLDs to increase lipolysis. Lipophagy–lipolysis crosstalk. Activation
of chaperone-mediated autophagy degrades the cLD coat proteins perilipin 2 and perilipin 3 through the coordinated
action of heat-shock cognate 71 kDa protein (HSC70; also known as HSPA8) and the receptor lysosome-associated
membrane glycoprotein 2A (LAMP2A). Removal of perilipins from the cLD surface allows the docking of autophagy
proteins, and the cLD surface becomes accessible to neutral lipolysis by ATGL in complex with lipid droplet-binding protein
CGI‑58, which hydrolyses the cLD triacylglycerols to generate FFAs. Acid lipolysis. After receptor-mediated endocytosis
of lipoproteins, cholesteryl esters and triacylglycerols are hydrolysed by LAL in lysosomes. ER, endoplasmic reticulum.

NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 18 | NOVEMBER 2017 | 679

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
REVIEWS

Fatty liver disease
A reversible condition
characterized by the excessive
accumulation of neutral lipids
in the liver. This condition
can be subdivided into
alcoholic fatty liver disease
and non-alcoholic fatty
liver disease.
than in mice lacking ATGL or its co-activator, CGI‑58.
For example, liver-­specific ATG7 deficiency, which is
expected to completely abolish hepatic autophagy, caused
a marked increase in hepatic fat content in some, but not
all, studies101,128–131. Conflicting results were also reported
for the role of autophagy in fatty liver disease. The majority
of studies132–134 provided evidence for a protective role of
lipophagy against hepatosteatosis, but others concluded
that hepatocyte-specific autophagy deficiency per se does
not ­exacerbate hepatic steatosis131,135.
Lipophagy and lipid synthesis. In addition to lipid catabo­
lism, lipophagy has important roles in adipogenesis, lipid
synthesis and cLD biogenesis. LC3‑II binding to cLDs
is required for lipid synthesis and cLD growth in liver
cells and cardiomyocytes, and defective LC3 conjugation
in mice with liver-specific ATG7 deficiency inhibited
cLD formation129,136. Similar results were observed in
Atg5−/− mouse embryonic fibroblasts137. Consistent with
a role for autophagy in adipogenesis, mice with adipose-­
specific ATG7 deficiency and newborn mice with global
ATG5 deficiency exhibit decreased adipose mass138–140,
the opposite of what would have been predicted from a
defect in lipophagic triacylglycerol catabolism. Autophagy
was found to be crucial for adipocyte differentiation at
early stages but dispensable later during differentiation141.
Finally, as mentioned above, autophagy-defective livers
are unable to store lipids in response to fasting-induced
WAT lipolysis131,135,136,142. Together, these data suggest that
besides having a role in catabolic pathways, autophagy
has a function in anabolic lipid metabolism. It is conceiv­
able that phosphatidylethanolamine-conjugated LC3‑II
(FIG. 4a) binds to cLDs and acts as a platform for both
­anabolic and catabolic processes.
Lipolysis–lipophagy crosstalk
The recent findings that metabolic kinases such as ERK
interact with LC3‑II143 and that LC3‑II binds ATGL on
cLDs in brown adipocytes after cold exposure to promote
triacylglycerol hydrolysis144 strengthened the possibility
of a functional link between autophagy and cytosolic
lipolysis. Clear discrimination between and elucid­
ation of the autophagy-related and autophagy-­unrelated
functions of ATGL will require systematic epistatic
experiments combined with autophagic flux analyses.
In addition to ATGL, CGI‑58 may be directly involved
in the regulation of autophagy by preventing cleavage of
beclin 1 by caspase 3 (REF. 145).
Further evidence for a functional link between auto­
phagy and lipolysis came from the discovery that CMA
degrades cLD-associated proteins and thereby regulates
neutral lipolysis146. In CMA, heat-shock cognate 71 kDa
protein recognizes and binds cytosolic proteins and inter­
acts with LAMP2A and other chaperones to form a trans­
location complex, which is transferred to lysosomes for
degradation (FIG. 4b). Perilipin 2 and perilipin 3, which are
abundant cLD-associated proteins that shield cLDs from
lipases and lipolysis, are CMA targets146. Consequently,
the removal of perilipin 2 and perilipin 3 by CMA ­enables
ATGL to efficiently access the cLD surface, thereby
increasing lipolytic rates.
A third form of crosstalk involves the consecutive
action of autophagy and neutral lipolysis147. In the first
step, autophagic digestion of membrane material and sub­
sequent triacylglycerol synthesis lead to the formation of
cLDs. In the second step, cLD-associated triacylglycerols
are hydrolysed by ATGL. This process is closely linked to
mitochondrial FFA uptake and oxidation.
Importantly, neutral lipolysis may be directly involved
in the regulation of autophagy, specifically through
the effect of ATGL on the hepatic function of PPARα
and SIRT1 (described above), both of which are well-­
established activators of autophagy in the liver 119,121.
Together with the co‑regulation of lipophagy and n ­ eutral
lipolysis by the major metabolic hormones and their
associ­ated regulatory hubs (mTOR, AMPK and the FOXO
transcription factors) (FIG. 3), these data support the view
that neutral lipolysis and lipophagy should not be con­
sidered distinct, but instead should be seen as two sides
of the same coin.
Therapeutic potential
The question of whether the pharmacological inhib­
ition of lipolysis in WAT and the concomitant decline in
plasma FFA concentrations represent a beneficial strat­
egy for preventing and treating ectopic lipid accumulation
and lipotoxicity has been widely discussed. The concept
appears attractive with regard to achieving a lasting reduc­
tion of hepatosteatosis and preventing its progression to
more morbid pathologies, such as steatohepatitis, cirrho­
sis and liver cancer 148. Decreased FFA concentrations
in the plasma may also increase insulin sensitivity and
decrease the concentration of atherogenic lipoproteins
(including VLDL)149. The antilipolytic action of nico­
tinic acid (­niacin), a drug long known to lower plasma
triacyl­glycerol levels and increase the concentrations of
high-density lipoprotein (HDL) cholesterol, provided
partial proof of this concept 150. Niacin binds to the adi­
pose tissue-specific G-protein coupled receptor 109A
(GPR109A; also known as HCAR2), leading to decreased
cellular cAMP concentrations and reduced lipolysis151.
The antilipolytic effect decreases plasma FFA concentra­
tions, which in turn results in a favourable plasma lipo­
protein profile owing to decreased VLDL production in
the liver and increased HDL-mediated reverse cholesterol
transport. Unfortunately, niacin has several adverse side
effects that restrict its broad application152. Additionally, in
Gpr109a−/− mice, the decrease in hepatic VLDL synthesis
and the lowering of plasma triacylglycerol concentrations
by ­niacin may not result from the inhibition of adipose
lipo­lysis, but rather from a direct effect of niacin on hepatic
­triacylglycerol and VLDL synthesis in hepatocytes152,153.
Another strategy for inhibiting lipolysis in WAT
focused on chemically synthesizing small-molecule inhib­
itors of HSL154,155; these compounds were developed
before the discovery of ATGL. HSL inhibitors moderately
decreased plasma FFA concentrations in treated rats and
mice. Importantly, however, HSL inhibition improved
insulin sensitivity, decreased plasma glucose levels and
reduced glucose-induced insulin secretion from pan­
creatic β-cells, in line with the concept that inhibition of
lipolysis may beneficially affect glucose homeostasis156.

680 | NOVEMBER 2017 | VOLUME 18 www.nature.com/nrm

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
 REVIEWS

Box 3 | Lipolysis in cancer and cancer-associated cachexia

The roles of lipolysis and lipophagy in tumorigenesis and the development
of cancer-associated cachexia (CAC) have only recently attracted
substantial attention. In 2010, monoacylglycerol lipase (MGL) was described
as a potential oncogene181. High MGL expression levels in cancer cells
promote cell proliferation and tumour growth and strongly correlate with
malignancy. Conversely, inhibition of MGL reduces cancer cell proliferation
and metastasis and induces cancer cell apoptosis182. The beneficial effects
of MGL inhibition probably involve the induction of endocannabinoid
signalling by inhibition of 2‑arachidonylglycerol hydrolysis and a decrease
in the levels of lipid mediators derived from free fatty acids (FFAs) owing to
impaired FFA production183.
How triacylglycerol hydrolysis by adipose triglyceride lipase (ATGL) and
lipid droplet-binding protein CGI‑58 affects cancer cell metabolism
and proliferation is less clear. A robust correlation between low ATGL
expression on the one hand and malignancy and reduced survival on
the other was reported for non-small-cell lung cancers, pancreatic
adenocarcinoma and leiomyosarcoma184. Similarly, the downregulation
of ATGL by epigenetic silencing of the gene encoding the E2 ligase UBCH8
(also known as UBE2L6) correlated with poor prognosis in nasopharyngeal
cancer185. Decreased UBCH8 expression leads to reduced ATGL stability
and activity and triacylglycerol accumulation185. In mice, ATGL deficiency
frequently leads to pulmonary neoplasia and adenocarcinoma184, and
deficiency of both ATGL and hormone-sensitive lipase (HSL) causes
liposarcoma186, suggesting a tumour suppressor function for these lipolytic
enzymes. The biological basis for the pro-tumorigenic activity of MGL
and the antitumorigenic activity of ATGL is not known.
The role of CGI‑58 in tumorigenesis also remains insufficiently
understood. Overexpression of CGI‑58 in macrophages causes increased
tumour growth in a xenotransplant model of colorectal cancer187.
Another study identified CGI‑58 as a potential tumour suppressor because
loss of CGI‑58 increased the propensity for tumour growth in a mouse
intestinal cancer model188. Several, mostly ATGL-independent mechanisms
were held responsible for various CGI‑58‑dependent cancer phenotypes,
including the regulation of AMP-activated protein kinase (AMPK), the
production of cellular polyamine and inhibition of autophagy by the
interaction of CGI‑58 with beclin 1 (REFS 145,187,188).
Several recent studies reported a tumour suppressor function for lysosomal
acid lipase (LAL). LAL deficiency causes tumour growth and metastasis in
various cancer models189. LAL deficiency leads to mTOR-mediated expansion
and organ infiltration of myeloid-derived suppressor cells (MDSCs), which
suppress tumour immune surveillance. Re‑expression of LAL in hepatocytes
or MDSCs reduces the abundance of MDSCs in the liver, improves hepatic
lipid metabolism, attenuates inflammation and decreases liver metastasis in
a mouse xenotransplant melanoma model190,191. Similarly, LAL re‑expression
in lung epithelial cells improved functional and metabolic parameters and
reduced the metastasis of lung cancer in mice192. Although still preliminary
and incomplete, these results highlight a previously underestimated role for
neutral and acid lipolysis in cancer cell metabolism and tumorigenesis.
Lipases involved in neutral lipolysis may also have an essential role in CAC.
Lipolysis is strongly induced in the white adipose tissue (WAT) of individuals
with CAC160–162, and deficiency of ATGL and HSL in mice partially protects
against CAC in two xenotransplant cancer models161. CAC is accompanied
by the activation of thermogenesis in brown adipose tissue193 and increased
‘browning’ of subcutaneous WAT194. Anti-inflammatory treatment partially
reversed these metabolic aberrations, suggesting that CAC predominantly
resulted from systemic inflammation194. A recent study demonstrated that
a pronounced downregulation of AMPK induced HSL and provoked CAC
in murine cancer models163.

Cachexia
A wasting syndrome
characterized by an
unintentional and nutritionally
irreversible loss of body mass
(muscle and fat mass). Various
chronic diseases can lead to
cachexia, but it is most
common in cancer.
Recently, an ATGL-specific inhibitor (atglistatin)
was discovered and tested in mice157. Atglistatin treat­
ment effectively lowered plasma FFA, triacylglycerol and
choles­terol concentrations; improved insulin sensitivity;
and attenuated hepatosteatosis in mice treated with a
high-fat diet158. Unexpectedly, atglistatin-treated a­ nimals
were resistant to obesity induced by a high-fat diet;
showed no signs of triacylglycerol deposition in ectopic
tissues; and had normal or even improved cardiac func­
tion. The drastic reduction in liver fat, with no signs of
lipid accumulation, in skeletal and cardiac muscles differs
strikingly from observations in Atgl−/− mice, which exhibit
systemic lipid accumulation10. Although the reasons for
these contrasting results are not completely understood,
it is conceivable that the competitive and reversible inhib­
itory properties of atglistatin lead to oscillating inhibition
of ATGL, thereby preventing the detrimental effects of
complete ATGL deficiency. The fact that atglistatin is
not detectably absorbed by cardiac or skeletal muscle
possibly contributes to the absence of muscle steatosis
in treated animals. Thus, the inhibition of lipolysis by
ATGL and/or HSL inhibitors may serve as a powerful
strategy in the prevention and treatment of metabolic
disease. In particu­lar, liver steatosis, with its tremendous
prevalence, represents an important target. Whether drug
treatment can reduce atherosclerotic plaque formation, as
observed in Atgl−/− mice159, needs to be examined.
Inhibitors of lipolysis may also be beneficial for
the treatment of cancer-associated cachexia (CAC).
In the past decade, the roles of neutral and acid lipases
in tumori­genesis and the pathogenesis of CAC have
attracted major interest (BOX 3). ATGL and HSL activity
is high in the WAT of cancer patients with CAC160–162 and
contributes to the associated wasting process161,163. ATGL
or HSL deficiency partially prevents CAC in murine
­cancer models161,163. Whether pharmacological inhibition
of lipolysis can prevent CAC in mice is currently being
investigated. The roles of autophagy and lipophagy in
CAC are unknown.
Conclusion and future perspective
Despite major progress in our understanding of how the
different branches of lipolysis function, how these path­
ways communicate with each other and how they affect
the (patho)physiology of mammals, many key questions
remain unanswered. (i) The structural and functional
characterization of the neutral lipolytic machinery
is incomplete, as the three-dimensional structures of
enzymes and regulators are not available, and the ATGL-
independent function(s) of CGI‑58 remain(s) elusive. (ii)
The extent to which lipophagy contributes to bulk triacyl­
glycerol catabolism in various cell types and tissues needs
to be determined. (iii) The multitude of biological pro­
cesses that are regulated by products of neutral lipolysis,
lipophagy and acid lipolysis require better characteriza­
tion. (iv) The mechanistic link between neutral lipolysis
and lipophagy requires further interrogation. (v) Finally,
we need to better understand the roles of neutral lipolysis,
lipophagy and acid lipolysis in the pathogenesis of meta­
bolic disorders and to evaluate whether the inhibition of
lipolysis can help to prevent insulin resistance and fatty
liver disease in humans.

NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 18 | NOVEMBER 2017 | 681

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
REVIEWS

1. Unger, R. H. Lipotoxic diseases. Annu. Rev. Med. 53,
319–336 (2002).
2. Armand, M. Lipases and lipolysis in the human
digestive tract: where do we stand? Curr. Opin. Clin.
Nutr. Metab. Care 10, 156–164 (2007).
3. Young, S. G. & Zechner, R. Biochemistry and
pathophysiology of intravascular and intracellular
lipolysis. Genes Dev. 27, 459–484 (2013).
4. Zechner, R. FAT FLUX: enzymes, regulators, and
pathophysiology of intracellular lipolysis. EMBO Mol.
Med. 7, 359–362 (2015).
5. Dubland, J. A. & Francis, G. A. Lysosomal acid lipase:
at the crossroads of normal and atherogenic
cholesterol metabolism. Front. Cell Dev. Biol. 3,
3 (2015).
6. Singh, R. et al. Autophagy regulates lipid metabolism.
Nature 458, 1131–1135 (2009).
7. Zimmermann, R. et al. Fat mobilization in adipose
tissue is promoted by adipose triglyceride lipase.
Science 306, 1383–1386 (2004).
8. Villena, J. A., Roy, S., Sarkadi-Nagy, E., Kim, K. H.
& Sul, H. S. Desnutrin, an adipocyte gene encoding
a novel patatin domain-containing protein, is induced
by fasting and glucocorticoids: ectopic expression
of desnutrin increases triglyceride hydrolysis. J. Biol.
Chem. 279, 47066–47075 (2004).
9. Jenkins, C. M. et al. Identification, cloning,
expression, and purification of three novel human
calcium-independent phospholipase A2 family
members possessing triacylglycerol lipase and
acylglycerol transacylase activities. J. Biol. Chem. 279,
48968–48975 (2004).
10. Haemmerle, G. et al. Defective lipolysis and altered
energy metabolism in mice lacking adipose triglyceride
lipase. Science 312, 734–737 (2006).
11. Vaughan, M., Berger, J. E. & Steinberg, D. Hormone-
sensitive lipase and monoglyceride lipase activities in
adipose tissue. J. Biol. Chem. 239, 401–409 (1964).
12. Lass, A. et al. Adipose triglyceride lipase-mediated
lipolysis of cellular fat stores is activated by CGI‑58
and defective in Chanarin-Dorfman Syndrome.
Cell Metab. 3, 309–319 (2006).
13. Yang, X. et al. The G0/G1 switch gene 2 regulates
adipose lipolysis through association with adipose
triglyceride lipase. Cell Metab. 11, 194–205 (2010).
14. Radner, F. P. et al. Growth retardation, impaired
triacylglycerol catabolism, hepatic steatosis, and lethal
skin barrier defect in mice lacking comparative gene
identification‑58 (CGI‑58). J. Biol. Chem. 285,
7300–7311 (2010).
15. Grond, S. et al. Skin barrier development depends
on CGI‑58 protein expression during late-stage
keratinocyte differentiation. J. Invest. Dermatol. 137,
403–413 (2017).
16. Grond, S. et al. PNPLA1 deficiency in mice and
humans leads to a defect in the synthesis of
omega‑O‑Acylceramides. J. Invest. Dermatol. 137,
394–402 (2017).
17. Stone, S. J. et al. Lipopenia and skin barrier
abnormalities in DGAT2‑deficient mice. J. Biol. Chem.
279, 11767–11776 (2004).
18. Russell, L. & Forsdyke, D. R. A human putative
lymphocyte G0/G1 switch gene containing a CpG-rich
island encodes a small basic protein with the potential
to be phosphorylated. DNA Cell Biol. 10, 581–591
(1991).
19. Cerk, I. K. et al. A peptide derived from G0/G1 switch
gene 2 acts as noncompetitive inhibitor of adipose
triglyceride lipase. J. Biol. Chem. 289, 32559–32570
(2014).
20. Lu, X., Yang, X. & Liu, J. Differential control of
ATGL-mediated lipid droplet degradation by CGI‑58
and G0S2. Cell Cycle 9, 2719–2725 (2010).
21. Heckmann, B. L. et al. Defective adipose lipolysis and
altered global energy metabolism in mice with adipose
overexpression of the lipolytic inhibitor G0/G1 switch
gene 2 (G0S2). J. Biol. Chem. 289, 1905–1916
(2014).
22. Zhang, X. et al. Targeted disruption of G0/G1 switch
gene 2 enhances adipose lipolysis, alters hepatic
energy balance, and alleviates high-fat diet-induced
liver steatosis. Diabetes 63, 934–946 (2014).
23. Wang, Y. et al. The G0/G1 switch gene 2 is an
important regulator of hepatic triglyceride
metabolism. PLoS ONE 8, e72315 (2013).
24. Heier, C. et al. G0/G1 switch gene 2 regulates cardiac
lipolysis. J. Biol. Chem. 290, 26141–26150 (2015).
25. Hofer, P. et al. Fatty acid-binding proteins interact
with comparative gene identification‑58 linking
lipolysis with lipid ligand shuttling. J. Biol. Chem. 290,
18438–18453 (2015).
26. Notari, L. et al. Identification of a lipase-linked cell
membrane receptor for pigment epithelium-derived
factor. J. Biol. Chem. 281, 38022–38037 (2006).
27. Chung, C. et al. Anti-angiogenic pigment epithelium-
derived factor regulates hepatocyte triglyceride
content through adipose triglyceride lipase (ATGL).
J. Hepatol. 48, 471–478 (2008).
28. Borg, M. L. et al. Pigment epithelium-derived factor
regulates lipid metabolism via adipose triglyceride
lipase. Diabetes 60, 1458–1466 (2011).
29. Singh, M. et al. Fat-specific protein 27 inhibits lipolysis
by facilitating the inhibitory effect of transcription
factor Egr1 on transcription of adipose triglyceride
lipase. J. Biol. Chem. 289, 14481–14487 (2014).
30. Grahn, T. H. et al. Fat-specific protein 27 (FSP27)
interacts with adipose triglyceride lipase (ATGL) to
regulate lipolysis and insulin sensitivity in human
adipocytes. J. Biol. Chem. 289, 12029–12039
(2014).
31. Eichmann, T. O. et al. Studies on the substrate
and stereo/regioselectivity of adipose triglyceride
lipase, hormone-sensitive lipase, and diacylglycerol-
O‑acyltransferases. J. Biol. Chem. 287, 41446–41457
(2012).
32. Rodriguez, J. A. et al. In vitro stereoselective
hydrolysis of diacylglycerols by hormone-sensitive
lipase. Biochim. Biophys. Acta 1801, 77–83 (2010).
33. Lafontan, M. & Langin, D. Lipolysis and lipid
mobilization in human adipose tissue. Prog. Lipid Res.
48, 275–297 (2009).
34. Fischer, J. et al. The gene encoding adipose
triglyceride lipase (PNPLA2) is mutated in neutral lipid
storage disease with myopathy. Nat. Genet. 39,
28–30 (2007).
35. Osuga, J. et al. Targeted disruption of hormone-
sensitive lipase results in male sterility and adipocyte
hypertrophy, but not in obesity. Proc. Natl Acad.
Sci. USA 97, 787–792 (2000).
36. Albert, J. S. et al. Null mutation in hormone-sensitive
lipase gene and risk of type 2 diabetes. N. Engl.
J. Med. 370, 2307–2315 (2014).
37. Petrosino, S. & Di Marzo, V. FAAH and MAGL
inhibitors: therapeutic opportunities from regulating
endocannabinoid levels. Curr. Opin. Investig. Drugs
11, 51–62 (2010).
38. Subramanian, V. et al. Perilipin A mediates the
reversible binding of CGI‑58 to lipid droplets in 3T3‑L1
adipocytes. J. Biol. Chem. 279, 42062–42071
(2004).
39. Granneman, J. G. & Moore, H. P. Location, location:
protein trafficking and lipolysis in adipocytes.
Trends Endocrinol. Metab. 19, 3–9 (2008).
40. Watt, M. J. & Steinberg, G. R. Regulation and function
of triacylglycerol lipases in cellular metabolism.
Biochem. J. 414, 313–325 (2008).
41. Collins, S. A heart-adipose tissue connection in the
regulation of energy metabolism. Nat. Rev. Endocrinol.
10, 157–163 (2014).
42. Kaltenecker, D. et al. Adipocyte STAT5 deficiency
promotes adiposity and impairs lipid mobilisation
in mice. Diabetologia 60, 296–330 (2016).
43. Saltiel, A. R. Insulin signaling in the control of glucose
and lipid homeostasis. Handb Exp. Pharmacol. 233,
51–71 (2016).
44. DiPilato, L. M. et al. The role of PDE3B
phosphorylation in the inhibition of lipolysis by insulin.
Mol. Cell. Biol. 35, 2752–2760 (2015).
45. Chakrabarti, P. & Kandror, K. V. FoxO1 controls
insulin-dependent adipose triglyceride lipase (ATGL)
expression and lipolysis in adipocytes. J. Biol. Chem.
284, 13296–13300 (2009).
46. Chakrabarti, P. et al. SIRT1 controls lipolysis in
adipocytes via FOXO1‑mediated expression of ATGL.
J. Lipid Res. 52, 1693–1701 (2011).
47. Picard, F. et al. Sirt1 promotes fat mobilization in
white adipocytes by repressing PPAR-γ. Nature 429,
771–776 (2004).
48. Lasa, A. et al. Resveratrol regulates lipolysis via
adipose triglyceride lipase. J. Nutr. Biochem. 23,
379–384 (2012).
49. Lamming, D. W. & Sabatini, D. M. A. Central role for
mTOR in lipid homeostasis. Cell Metab. 18, 465–469
(2013).
50. Ceddia, R. B. The role of AMP-activated protein kinase
in regulating white adipose tissue metabolism.
Mol. Cell Endocrinol. 366, 194–203 (2013).
51. Chakrabarti, P., English, T., Shi, J., Smas, C. M.
& Kandror, K. V. Mammalian target of rapamycin
complex 1 suppresses lipolysis, stimulates lipogenesis,
and promotes fat storage. Diabetes 59, 775–781
(2010).
52. Chakrabarti, P. et al. Insulin inhibits lipolysis in
adipocytes via the evolutionarily conserved
mTORC1‑Egr1‑ATGL-mediated pathway. Mol. Cell.
Biol. 33, 3659–3666 (2013).
53. Kumar, A. et al. Fat cell-specific ablation of rictor
in mice impairs insulin-regulated fat cell and whole-
body glucose and lipid metabolism. Diabetes 59,
1397–1406 (2010).
54. Gaidhu, M. P. et al. Prolonged AICAR-induced
AMP‑kinase activation promotes energy dissipation
in white adipocytes: novel mechanisms integrating
HSL and ATGL. J. Lipid Res. 50, 704–715 (2009).
55. Kim, S. J. et al. AMPK phosphorylates desnutrin/ATGL
and hormone-sensitive lipase to regulate lipolysis
and fatty acid oxidation within adipose tissue.
Mol. Cell. Biol. 36, 1961–1976 (2016).
56. Watt, M. J. et al. Regulation of HSL serine
phosphorylation in skeletal muscle and adipose tissue.
Am. J. Physiol. Endocrinol. Metab. 290, E500–E508
(2006).
57. Ahmadian, M. et al. Desnutrin/ATGL is regulated by
AMPK and is required for a brown adipose phenotype.
Cell Metab. 13, 739–748 (2011).
58. Pagnon, J. et al. Identification and functional
characterization of protein kinase A phosphorylation
sites in the major lipolytic protein, adipose
triglyceride lipase. Endocrinology 153, 4278–4289
(2012).
59. Karlsson, M. et al. Exon-intron organization and
chromosomal localization of the mouse monoglyceride
lipase gene. Gene 272, 11–18 (2001).
60. Rakhshandehroo, M. et al. Comprehensive analysis
of PPARα-dependent regulation of hepatic lipid
metabolism by expression profiling. PPAR Res. 2007,
26839 (2007).
61. Haemmerle, G. et al. ATGL-mediated fat catabolism
regulates cardiac mitochondrial function via PPAR-α
and PGC‑1. Nat. Med. 17, 1076–1085 (2011).
62. Zierler, K. A. et al. Functional cardiac lipolysis in mice
critically depends on comparative gene
identification‑58. J. Biol. Chem. 288, 9892–9904
(2013).
63. Schoiswohl, G. et al. Adipose triglyceride lipase plays
a key role in the supply of the working muscle with
fatty acids. J. Lipid Res. 51, 490–499 (2010).
64. Wu, J. W. et al. Deficiency of liver adipose triglyceride
lipase in mice causes progressive hepatic steatosis.
Hepatology 54, 122–132 (2011).
65. Ong, K. T., Mashek, M. T., Bu, S. Y., Greenberg, A. S.
& Mashek, D. G. Adipose triglyceride lipase is a major
hepatic lipase that regulates triacylglycerol turnover
and fatty acid signaling and partitioning. Hepatology
53, 116–126 (2011).
66. Obrowsky, S. et al. Adipose triglyceride lipase is
a TG hydrolase of the small intestine and regulates
intestinal PPARα signaling. J. Lipid Res. 54, 425–435
(2013).
67. Chandak, P. G. et al. Efficient phagocytosis requires
triacylglycerol hydrolysis by adipose triglyceride lipase.
J. Biol. Chem. 285, 20192–20201 (2010).
68. Mottillo, E. P., Bloch, A. E., Leff, T. & Granneman, J. G.
Lipolytic products activate peroxisome proliferator-
activated receptor (PPAR) α and δ in brown adipocytes
to match fatty acid oxidation with supply. J. Biol.
Chem. 287, 25038–25048 (2012).
69. Schoiswohl, G. et al. Impact of reduced ATGL-mediated
adipocyte lipolysis on obesity-associated insulin
resistance and inflammation in male mice.
Endocrinology 156, 3610–3624 (2015).
70. Jaeger, D. et al. Fasting-induced G0/G1 switch gene 2
and FGF21 expression in the liver are under regulation
of adipose tissue derived fatty acids. J. Hepatol. 63,
437–445 (2015).
71. Tang, T. et al. Desnutrin/ATGL activates PPARδ to
promote mitochondrial function for insulin secretion
in islet beta cells. Cell Metab. 18, 883–895 (2013).
72. Schreiber, R. et al. Hypophagia and metabolic
adaptations in mice with defective ATGL-mediated
lipolysis cause resistance to HFD-induced obesity.
Proc. Natl Acad. Sci. USA 112, 13850–13855
(2015).
73. Haemmerle, G. et al. Hormone-sensitive lipase
deficiency in mice causes diglyceride accumulation in
adipose tissue, muscle, and testis. J. Biol. Chem. 277,
4806–4815 (2002).
74. Armstrong, E. H., Goswami, D., Griffin, P. R., Noy, N.
& Ortlund, E. A. Structural basis for ligand regulation
of the fatty acid-binding protein 5, peroxisome
proliferator-activated receptor beta/delta
(FABP5‑PPARβ/δ) signaling pathway. J. Biol. Chem.
289, 14941–14954 (2014).

682 | NOVEMBER 2017 | VOLUME 18 www.nature.com/nrm

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
 REVIEWS

75. Tan, N. S. et al. Selective cooperation between fatty
acid binding proteins and peroxisome proliferator-
activated receptors in regulating transcription.
Mol. Cell. Biol. 22, 5114–5127 (2002).
76. Wolfrum, C., Borrmann, C. M., Borchers, T.
& Spener, F. Fatty acids and hypolipidemic drugs
regulate peroxisome proliferator-activated receptors
α- and γ-mediated gene expression via liver fatty acid
binding protein: a signaling path to the nucleus.
Proc. Natl Acad. Sci. USA 98, 2323–2328 (2001).
77. Shen, W. J., Sridhar, K., Bernlohr, D. A. & Kraemer, F.
B. Interaction of rat hormone-sensitive lipase with
adipocyte lipid-binding protein. Proc. Natl Acad.
Sci. USA 96, 5528–5532 (1999).
78. Smith, A. J. et al. Physical association between the
adipocyte fatty acid-binding protein and hormone-
sensitive lipase: a fluorescence resonance energy
transfer analysis. J. Biol. Chem. 279, 52399–52405
(2004).
79. Khan, S. A. et al. ATGL-catalyzed lipolysis regulates
SIRT1 to control PGC‑1γ/PPAR-α signaling. Diabetes
64, 418–426 (2015).
80. Ng, F. & Tang, B. L. Sirtuins’ modulation of autophagy.
J. Cell. Physiol. 228, 2262–2270 (2013).
81. Lettieri Barbato, D., Tatulli, G., Aquilano, K.
& Ciriolo, M. R. FoxO1 controls lysosomal acid lipase
in adipocytes: implication of lipophagy during nutrient
restriction and metformin treatment. Cell Death Dis.
4, e861 (2013).
82. Wagner, G. R. & Payne, R. M. Widespread and
enzyme-independent Nepsilon-acetylation and
Nepsilon-succinylation of proteins in the chemical
conditions of the mitochondrial matrix. J. Biol. Chem.
288, 29036–29045 (2013).
83. Weinert, B. T. et al. Acetylation dynamics and
stoichiometry in Saccharomyces cerevisiae. Mol. Syst.
Biol. 10, 716 (2014).
84. Weinert, B. T., Moustafa, T., Iesmantavicius, V.,
Zechner, R. & Choudhary, C. Analysis of acetylation
stoichiometry suggests that SIRT3 repairs nonenzymatic
acetylation lesions. EMBO J. 34, 2620–2632 (2015).
85. Eisenberg, T. et al. Nucleocytosolic depletion of the
energy metabolite acetyl-coenzyme a stimulates
autophagy and prolongs lifespan. Cell Metab. 19,
431–444 (2014).
86. Marino, G. et al. Regulation of autophagy by cytosolic
acetyl-coenzyme A. Mol. Cell 53, 710–725 (2014).
87. Pougovkina, O. et al. Mitochondrial protein
acetylation is driven by acetyl-CoA from fatty acid
oxidation. Hum. Mol. Genet. 23, 3513–3522 (2014).
88. Hebert, A. S. et al. Calorie restriction and SIRT3
trigger global reprogramming of the mitochondrial
protein acetylome. Mol. Cell 49, 186–199 (2013).
89. Hirschey, M. D. et al. SIRT3 regulates mitochondrial
fatty-acid oxidation by reversible enzyme
deacetylation. Nature 464, 121–125 (2010).
90. Perry, R. J. et al. Hepatic acetyl CoA links adipose
tissue inflammation to hepatic insulin resistance
and type 2 diabetes. Cell 160, 745–758 (2015).
91. Sheriff, S., Du, H. & Grabowski, G. A. Characterization
of lysosomal acid lipase by site-directed mutagenesis
and heterologous expression. J. Biol. Chem. 270,
27766–27772 (1995).
92. Warner, T. G., Dambach, L. M., Shin, J. H.
& O’Brien, J. S. Purification of the lysosomal acid
lipase from human liver and its role in lysosomal lipid
hydrolysis. J. Biol. Chem. 256, 2952–2957 (1981).
93. Grumet, L. et al. Lysosomal acid lipase hydrolyzes
retinyl ester and affects retinoid turnover. J. Biol.
Chem. 291, 17977–17987 (2016).
94. Sando, G. N. & Henke, V. L. Recognition and receptor-
mediated endocytosis of the lysosomal acid lipase
secreted by cultured human fibroblasts. J. Lipid Res.
23, 114–123 (1982).
95. Haka, A. S. et al. Macrophages create an acidic
extracellular hydrolytic compartment to digest
aggregated lipoproteins. Mol. Biol. Cell 20,
4932–4940 (2009).
96. Goldstein, J. L. & Brown, M. S. A century of
cholesterol and coronaries: from plaques to genes
to statins. Cell 161, 161–172 (2015).
97. Dieckmann, M., Dietrich, M. F. & Herz, J. Lipoprotein
receptors—an evolutionarily ancient multifunctional
receptor family. Biol. Chem. 391, 1341–1363 (2010).
98. Du, H., Duanmu, M., Witte, D. & Grabowski, G. A.
Targeted disruption of the mouse lysosomal acid lipase
gene: long-term survival with massive cholesteryl ester
and triglyceride storage. Hum. Mol. Genet. 7,
1347–1354 (1998).
99. Du, H. et al. Lysosomal acid lipase-deficient mice:
depletion of white and brown fat, severe
hepatosplenomegaly, and shortened life span. J. Lipid
Res. 42, 489–500 (2001).
100. Radovic, B. et al. Lysosomal acid lipase regulates VLDL
synthesis and insulin sensitivity in mice. Diabetologia
59, 1743–1752 (2016).
101. Settembre, C. et al. TFEB controls cellular lipid
metabolism through a starvation-induced
autoregulatory loop. Nat. Cell Biol. 15, 647–658
(2013).
102. Martina, J. A. et al. The nutrient-responsive
transcription factor TFE3 promotes autophagy,
lysosomal biogenesis, and clearance of cellular debris.
Sci. Signal 7, ra9 (2014).
103. Raben, N. & Puertollano, R. TFEB and TFE3: linking
lysosomes to cellular adaptation to stress. Annu. Rev.
Cell Dev. Biol. 32, 255–278 (2016).
104. O’Rourke, E. J. & Ruvkun, G. MXL‑3 and HLH‑30
transcriptionally link lipolysis and autophagy to
nutrient availability. Nat. Cell Biol. 15, 668–676
(2013).
105. Emanuel, R. et al. Induction of lysosomal biogenesis in
atherosclerotic macrophages can rescue lipid-induced
lysosomal dysfunction and downstream sequelae.
Arterioscler. Thromb. Vasc. Biol. 34, 1942–1952
(2014).
106. Cox, B. E., Griffin, E. E., Ullery, J. C. & Jerome, W. G.
Effects of cellular cholesterol loading on macrophage
foam cell lysosome acidification. J. Lipid Res. 48,
1012–1021 (2007).
107. Li, W., Yuan, X. M., Olsson, A. G. & Brunk, U. T.
Uptake of oxidized LDL by macrophages results in
partial lysosomal enzyme inactivation and relocation.
Arterioscler Thromb. Vasc. Biol. 18, 177–184
(1998).
108. Griffin, E. E., Ullery, J. C., Cox, B. E. & Jerome, W. G.
Aggregated LDL and lipid dispersions induce lysosomal
cholesteryl ester accumulation in macrophage foam
cells. J. Lipid Res. 46, 2052–2060 (2005).
109. Bowden, K. L. et al. Lysosomal acid lipase deficiency
impairs regulation of ABCA1 gene and formation
of high density lipoproteins in cholesteryl ester
storage disease. J. Biol. Chem. 286, 30624–30635
(2011).
110. Ikonen, E. Cellular cholesterol trafficking and
compartmentalization. Nat. Rev. Mol. Cell Biol. 9,
125–138 (2008).
111. Li, J. & Pfeffer, S. R. Lysosomal membrane
glycoproteins bind cholesterol and contribute to
lysosomal cholesterol export. eLife 5, e21635
(2016).
112. Chen, F. W., Gordon, R. E. & Ioannou, Y. A. NPC1 late
endosomes contain elevated levels of non-esterified
(‘free’) fatty acids and an abnormally glycosylated form
of the NPC2 protein. Biochem. J. 390, 549–561
(2005).
113. Passeggio, J. & Liscum, L. Flux of fatty acids through
NPC1 lysosomes. J. Biol. Chem. 280, 10333–10339
(2005).
114. Lee, S. D. & Tontonoz, P. Liver X receptors at the
intersection of lipid metabolism and atherogenesis.
Atherosclerosis 242, 29–36 (2015).
115. Folick, A. et al. Aging. Lysosomal signaling molecules
regulate longevity in Caenorhabditis elegans. Science
347, 83–86 (2015).
116. Levine, B. & Kroemer, G. Autophagy in the
pathogenesis of disease. Cell 132, 27–42 (2008).
117. Mizushima, N. & Komatsu, M. Autophagy: renovation
of cells and tissues. Cell 147, 728–741 (2011).
118. Lamb, C. A., Yoshimori, T. & Tooze, S. A. The
autophagosome: origins unknown, biogenesis
complex. Nat. Rev. Mol. Cell Biol. 14, 759–774
(2013).
119. Efeyan, A., Comb, W. C. & Sabatini, D. M.
Nutrient‑sensing mechanisms and pathways.
Nature 517, 302–310 (2015).
120. Feng, Y., Yao, Z. & Klionsky, D. J. How to control
self‑digestion: transcriptional, post-transcriptional,
and post-translational regulation of autophagy.
Trends Cell Biol. 25, 354–363 (2015).
121. Lee, J. M. et al. Nutrient-sensing nuclear receptors
coordinate autophagy. Nature 516, 112–115 (2014).
122. Ao, X., Zou, L. & Wu, Y. Regulation of autophagy
by the Rab GTPase network. Cell Death Differ. 21,
348–358 (2014).
123. Schroeder, B. et al. The small GTPase Rab7 as
a central regulator of hepatocellular lipophagy.
Hepatology 61, 1896–1907 (2015).
124. Itakura, E., Kishi-Itakura, C. & Mizushima, N.
The hairpin-type tail-anchored SNARE syntaxin 17
targets to autophagosomes for fusion with
endosomes/lysosomes. Cell 151, 1256–1269 (2012).
125. Saftig, P., Beertsen, W. & Eskelinen, E. L. LAMP‑2:
a control step for phagosome and autophagosome
maturation. Autophagy 4, 510–512 (2008).
126. Mijaljica, D., Prescott, M. & Devenish, R. J.
Microautophagy in mammalian cells: revisiting a
40‑year-old conundrum. Autophagy 7, 673–682
(2011).
127. Cingolani, F. & Czaja, M. J. Regulation and functions
of autophagic lipolysis. Trends Endocrinol. Metab. 27,
696–705 (2016).
128. Komatsu, M. et al. Impairment of starvation-induced
and constitutive autophagy in Atg7‑deficient mice.
J. Cell Biol. 169, 425–434 (2005).
129. Shibata, M. et al. The MAP1‑LC3 conjugation system
is involved in lipid droplet formation. Biochem.
Biophys. Res. Commun. 382, 419–423 (2009).
130. Conlon, D. M. et al. Inhibition of apolipoprotein B
synthesis stimulates endoplasmic reticulum
autophagy that prevents steatosis. J. Clin. Invest.
126, 3852–3867 (2016).
131. Kwanten, W. J. et al. Hepatocellular autophagy
modulates the unfolded protein response and fasting-
induced steatosis in mice. Am. J. Physiol. Gastrointest.
Liver Physiol. 311, G599–G609 (2016).
132. Mao, Y. et al. Ghrelin attenuated lipotoxicity via
autophagy induction and nuclear factor-κB inhibition.
Cell Physiol. Biochem. 37, 563–576 (2015).
133. Wang, Y., Singh, R., Xiang, Y. & Czaja, M. J.
Macroautophagy and chaperone-mediated autophagy
are required for hepatocyte resistance to oxidant
stress. Hepatology 52, 266–277 (2010).
134. Papackova, Z., Dankova, H., Palenickova, E.,
Kazdova, L. & Cahova, M. Effect of short- and long-
term high-fat feeding on autophagy flux and lysosomal
activity in rat liver. Physiol. Res. 61 (Suppl. 2),
S67–S76 (2012).
135. Ma, D. et al. Autophagy deficiency by hepatic FIP200
deletion uncouples steatosis from liver injury in
NAFLD. Mol. Endocrinol. 27, 1643–1654 (2013).
136. Shibata, M. et al. LC3, a microtubule-associated
protein1A/B light chain3, is involved in cytoplasmic
lipid droplet formation. Biochem. Biophys. Res.
Commun. 393, 274–279 (2010).
137. Yang, L., Li, P., Fu, S., Calay, E. S. & Hotamisligil, G. S.
Defective hepatic autophagy in obesity promotes ER
stress and causes insulin resistance. Cell Metab. 11,
467–478 (2010).
138. Baerga, R., Zhang, Y., Chen, P. H., Goldman, S.
& Jin, S. Targeted deletion of autophagy-related 5
(atg5) impairs adipogenesis in a cellular model
and in mice. Autophagy 5, 1118–1130 (2009).
139. Singh, R. et al. Autophagy regulates adipose mass
and differentiation in mice. J. Clin. Invest. 119,
3329–3339 (2009).
140. Zhang, Y. et al. Adipose-specific deletion of
autophagy‑related gene 7 (atg7) in mice reveals a
role in adipogenesis. Proc. Natl Acad. Sci. USA 106,
19860–19865 (2009).
141. Guo, L. et al. Transactivation of Atg4b by C/EBPbeta
promotes autophagy to facilitate adipogenesis.
Mol. Cell. Biol. 33, 3180–3190 (2013).
142. Takagi, A. et al. Mammalian autophagy is essential
for hepatic and renal ketogenesis during starvation.
Sci. Rep. 6, 18944 (2016).
143. Martinez-Lopez, N., Athonvarangkul, D., Mishall, P.,
Sahu, S. & Singh, R. Autophagy proteins regulate
ERK phosphorylation. Nat. Commun. 4, 2799
(2013).
144. Martinez-Lopez, N. et al. Autophagy in the CNS
and periphery coordinate lipophagy and lipolysis
in the brown adipose tissue and liver. Cell Metab.
23, 113–127 (2016).
145. Peng, Y. et al. ABHD5 interacts with BECN1 to
regulate autophagy and tumorigenesis of colon cancer
independent of PNPLA2. Autophagy 12, 2167–2182
(2016).
146. Kaushik, S. & Cuervo, A. M. Degradation of lipid
droplet-associated proteins by chaperone-mediated
autophagy facilitates lipolysis. Nat. Cell Biol. 17,
759–770 (2015).
147. Rambold, A. S., Cohen, S. & Lippincott-Schwartz, J.
Fatty acid trafficking in starved cells: regulation by
lipid droplet lipolysis, autophagy, and mitochondrial
fusion dynamics. Dev. Cell 32, 678–692 (2015).
148. Fabbrini, E., Sullivan, S. & Klein, S. Obesity and
nonalcoholic fatty liver disease: biochemical,
metabolic, and clinical implications. Hepatology 51,
679–689 (2010).
149. Samuel, V. T. & Shulman, G. I. The pathogenesis of
insulin resistance: integrating signaling pathways and
substrate flux. J. Clin. Invest. 126, 12–22 (2016).

NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 18 | NOVEMBER 2017 | 683

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.
REVIEWS

150. Ginsberg, H. N. & Reyes-Soffer, G. Niacin: a long
history, but a questionable future. Curr. Opin. Lipidol
24, 475–479 (2013).
151. Tunaru, S. et al. PUMA‑G and HM74 are receptors
for nicotinic acid and mediate its anti-lipolytic effect.
Nat. Med. 9, 352–355 (2003).
152. Lauring, B. et al. Niacin lipid efficacy is independent of
both the niacin receptor GPR109A and free fatty acid
suppression. Sci. Transl. Med. 4, 148ra115 (2012).
153. Ganji, S. H. et al. Niacin noncompetitively inhibits
DGAT2 but not DGAT1 activity in HepG2 cells. J. Lipid
Res. 45, 1835–1845 (2004).
154. Claus, T. H. et al. Specific inhibition of hormone-
sensitive lipase improves lipid profile while reducing
plasma glucose. J. Pharmacol. Exp. Ther. 315,
1396–1402 (2005).
155. Ebdrup, S., Refsgaard, H. H., Fledelius, C.
& Jacobsen, P. Synthesis and structure-activity
relationship for a novel class of potent and selective
carbamate-based inhibitors of hormone selective
lipase with acute in vivo antilipolytic effects. J. Med.
Chem. 50, 5449–5456 (2007).
156. Girousse, A. et al. Partial inhibition of adipose tissue
lipolysis improves glucose metabolism and insulin
sensitivity without alteration of fat mass. PLoS Biol.
11, e1001485 (2013).
157. Mayer, N. et al. Development of small-molecule
inhibitors targeting adipose triglyceride lipase.
Nat. Chem. Biol. 9, 785–787 (2013).
158. Schweiger, M. et al. Pharmacological inhibition of
adipose triglyceride lipase corrects high-fat diet-
induced insulin resistance and hepatosteatosis in mice.
Nat. Commun. 8, 14859 (2017).
159. Lammers, B. et al. Macrophage adipose triglyceride
lipase deficiency attenuates atherosclerotic lesion
development in low-density lipoprotein receptor
knockout mice. Arterioscler Thromb. Vasc. Biol. 31,
67–73 (2011).
160. Agustsson, T. et al. Mechanism of increased lipolysis in
cancer cachexia. Cancer Res. 67, 5531–5537 (2007).
161. Das, S. K. et al. Adipose triglyceride lipase
contributes to cancer-associated cachexia. Science
333, 233–238 (2011).
162. Ryden, M. et al. Lipolysis—not inflammation, cell
death, or lipogenesis—is involved in adipose tissue loss
in cancer cachexia. Cancer 113, 1695–1704 (2008).
163. Rohm, M. et al. An AMP-activated protein kinase-
stabilizing peptide ameliorates adipose tissue wasting
in cancer cachexia in mice. Nat. Med. 22, 1120–1130
(2016).
164. Massa, R. et al. Neutral lipid-storage disease with
myopathy and extended phenotype with novel
PNPLA2 mutation. Muscle Nerve 53, 644–648
(2016).
165. Natali, A. et al. Metabolic consequences of adipose
triglyceride lipase deficiency in humans: an in vivo
study in patients with neutral lipid storage disease
with myopathy. J. Clin. Endocrinol. Metab. 98,
E1540–E1548 (2013).
166. Dorfman, M. L., Hershko, C., Eisenberg, S.
& Sagher, F. Ichthyosiform dermatosis with systemic
lipidosis. Arch. Dermatol. 110, 261–266 (1974).
167. Chanarin, I. et al. Neutral-lipid storage disease: a new
disorder of lipid metabolism. Br. Med. J. 1, 553–555
(1975).
168. Lefevre, C. et al. Mutations in CGI‑58, the gene
encoding a new protein of the esterase/lipase/
thioesterase subfamily, in Chanarin-Dorfman syndrome.
Am. J. Hum. Genet. 69, 1002–1012 (2001).
169. Uchida, Y. et al. Neutral lipid storage leads to
acylceramide deficiency, likely contributing to the
pathogenesis of Dorfman-Chanarin syndrome.
J. Invest. Dermatol. 130, 2497–2499 (2010).
170. Zhang, J. et al. Comparative gene identification‑58
(CGI‑58) promotes autophagy as a putative
lysophosphatidylglycerol acyltransferase. J. Biol.
Chem. 289, 33044–33053 (2014).
171. Ghosh, A. K., Ramakrishnan, G., Chandramohan, C.
& Rajasekharan, R. CGI‑58, the causative gene
for Chanarin-Dorfman syndrome, mediates
acylation of lysophosphatidic acid. J. Biol. Chem. 283,
24525–24533 (2008).
172. Farhan, S. M. et al. A novel LIPE nonsense mutation
found using exome sequencing in siblings with late-
onset familial partial lipodystrophy. Can. J. Cardiol.
30, 1649–1654 (2014).
173. Wang, S. P. et al. The catalytic function of hormone-
sensitive lipase is essential for fertility in male mice.
Endocrinology 155, 3047–3053 (2014).
174. Abramov, A., Schorr, S. & Wolman, M. Generalized
xanthomatosis with calcified adrenals. AMA J. Dis.
Child 91, 282–286 (1956).
175. Sloan, H. R. & Fredrickson, D. S. Enzyme deficiency
in cholesteryl ester storage disease. J. Clin. Invest. 51,
1923–1926 (1972).
176. Burke, J. A. & Schubert, W. K. Deficient activity of acid
lipase in cholesterol-ester storage disease. J. Lab Clin.
Med. 78, 988–989 (1971).
177. Patrick, A. D. & Lake, B. D. Deficiency of an acid
lipase in Wolman’s disease. Nature 222, 1067–1068
(1969).
178. Bernstein, D. L., Hulkova, H., Bialer, M. G.
& Desnick, R. J. Cholesteryl ester storage disease:
review of the findings in 135 reported patients with an
underdiagnosed disease. J. Hepatol. 58, 1230–1243
(2013).
179. Hoffman, E. P., Barr, M. L., Giovanni, M. A.
& Murray, M. F. Lysosomal Acid Lipase Deficiency
(eds Pagon, R. A. et al.) (University of Washington,
2016).
180. Burton, B. K. et al. A phase 3 trial of sebelipase alfa
in lysosomal acid lipase deficiency. N. Engl. J. Med.
373, 1010–1020 (2015).
181. Nomura, D. K. et al. Monoacylglycerol lipase regulates
a fatty acid network that promotes cancer
pathogenesis. Cell 140, 49–61 (2010).
182. Zhang, J. et al. Monoacylglycerol lipase: A novel
potential therapeutic target and prognostic indicator
for hepatocellular carcinoma. Sci. Rep. 6, 35784
(2016).
183. Nomura, D. K. et al. Monoacylglycerol lipase exerts
dual control over endocannabinoid and fatty acid
pathways to support prostate cancer. Chem. Biol. 18,
846–856 (2011).
184. Al‑Zoughbi, W. et al. Loss of adipose triglyceride lipase
is associated with human cancer and induces mouse
pulmonary neoplasia. Oncotarget 7, 33832–33840
(2016).
185. Zhou, X. et al. Epigenetic downregulation of the
ISG15‑conjugating enzyme UbcH8 impairs lipolysis
and correlates with poor prognosis in
nasopharyngeal carcinoma. Oncotarget 6,
41077–41091 (2015).
186. Wu, J. W. et al. Epistatic interaction between
the lipase-encoding genes Pnpla2 and Lipe causes
liposarcoma in mice. PLoS Genet. 13, e1006716
(2017).
187. Miao, H. et al. Macrophage ABHD5 promotes
colorectal cancer growth by suppressing spermidine
production by SRM. Nat. Commun. 7, 11716 (2016).
188. Ou, J. et al. Loss of abhd5 promotes colorectal tumor
development and progression by inducing aerobic
glycolysis and epithelial-mesenchymal transition.
Cell Rep. 9, 1798–1811 (2014).
189. Yan, C., Zhao, T. & Du, H. Lysosomal acid lipase in
cancer. Oncoscience 2, 727–728 (2015).
190. Zhao, T., Du, H., Ding, X., Walls, K. & Yan, C.
Activation of mTOR pathway in myeloid-derived
suppressor cells stimulates cancer cell proliferation
and metastasis in lal−/− mice. Oncogene 34,
1938–1948 (2015).
191. Du, H., Zhao, T., Ding, X. & Yan, C. Hepatocyte-specific
expression of human lysosome acid lipase corrects
liver inflammation and tumor metastasis in lal−/− mice.
Am. J. Pathol. 185, 2379–2389 (2015).
192. Zhao, T., Ding, X., Du, H. & Yan, C. Lung epithelial
cell-specific expression of human lysosomal acid lipase
ameliorates lung inflammation and tumor metastasis
in lipa−/− mice. Am. J. Pathol. 186, 2183–2192
(2016).
193. Tsoli, M. et al. Activation of thermogenesis in brown
adipose tissue and dysregulated lipid metabolism
associated with cancer cachexia in mice. Cancer Res.
72, 4372–4382 (2012).
194. Petruzzelli, M. et al. A switch from white to brown fat
increases energy expenditure in cancer-associated
cachexia. Cell Metab. 20, 433–447 (2014).
Acknowledgements
Financial support was provided by the European Research
Council ERC Grant Agreement 340896, LipoCheX (R.Z.), the
DKs Molecular Enzymology (W901) (R.Z.) and Metabolic and
Cardiovascular Disease (W1226) (F.M., D.K.), the SFB
Lipotox (F30) funded by the Austrian Science Fund (FWF)
(R.Z., F.M., D.K.), the Louis-Jeantet Foundation (R.Z.), the
Fondation Leducq (grant 12CVD04) (R.Z.) and the
BioTechMed-Graz flagship projects EPIAge (F.M.) and Lipid
Signalling (D.K.). F.M. acknowledges additional support
from NAWI Graz; BioTechMed-Graz (“EPIAge”); FWF grants
P29262, P29203 and P27893; and the BMWFW and
University of Graz grants “Unkonventionelle Forschung”
and “Flysleep”.
Author contributions
Researching the literature for the article: R.Z., F.M. and D.K.;
substantial contributions to discussion of the content: R.Z.,
F.M. and D.K.; writing: R.Z. and D.K.; review and/or editing of
the manuscript before submission: R.Z., F.M. and D.K.
Competing interests statement
The authors declare no competing interests.
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional
claims in published maps and institutional affiliations.

684 | NOVEMBER 2017 | VOLUME 18 www.nature.com/nrm

©
2
0
1
7
M
a
c
m
i
l
l
a
n
P
u
b
l
i
s
h
e
r
s
L
i
m
i
t
e
d
,
p
a
r
t
o
f
S
p
r
i
n
g
e
r
N
a
t
u
r
e
.
A
l
l
r
i
g
h
t
s
r
e
s
e
r
v
e
d
.