Mammalian Sterols Novel Biological Roles of Cholesterol

Synthesis Intermediates, Oxysterols and Bile Acids - 1st

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Editors
Damjana Rozman Rolf Gebhardt
Center for Functional Genomics and Rudolf Schönheimer Institute of Biochemistry,
Bio-Chips Department of General Biochemistry
Institute of Biochemistry, Faculty of University of Leipzig, Faculty of Medicine
Medicine, University of Ljubljana Leipzig, Germany
Ljubljana, Slovenia

ISBN 978-3-030-39683-1 ISBN 978-3-030-39684-8 (eBook)

https://doi.org/10.1007/978-3-030-39684-8

# Springer Nature Switzerland AG 2020

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Contents

Sterols from the Post-Lanosterol Part of Cholesterol Synthesis:

Novel Signaling Players . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Cene Skubic and Damjana Rozman
Genetic Variability in Cholesterol Metabolism . . . . . . . . . . . . . . . . . . . . 23
Caitlin J. Smith, John M. Dagle, and Kelli K. Ryckman
Side-Chain Oxidized Oxysterols in Health and Disease . . . . . . . . . . . . . . 41
Ingemar Björkhem and Ulf Diczfalusy
Bile Acids and TGR5 (Gpbar1) Signaling . . . . . . . . . . . . . . . . . . . . . . . . 81
Verena Keitel, Christoph G. W. Gertzen, Sven Schäfer, Caroline Klindt,
Christina Wöhler, Kathleen Deutschmann, Maria Reich, Holger Gohlke,
and Dieter Häussinger
Bile Acids as Regulatory Signalling Molecules . . . . . . . . . . . . . . . . . . . . 101
Madlen Matz-Soja
Oxysterols and Bile Acid Act as Signaling Molecules That Regulate
Cholesterol Homeostasis: Nuclear Receptors LXR, FXR,
and Fibroblast Growth Factor 15/19 . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117
Klementina Fon Tacer
Cytochrome P450 Metabolism Leads to Novel Biological Sterols
and Other Steroids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145
F. Peter Guengerich and Francis K. Yoshimoto

v
Sterols from the Post-Lanosterol Part
of Cholesterol Synthesis: Novel Signaling
Players

Cene Skubic and Damjana Rozman

Introduction

Cholesterol is the major sterol in all mammalian cells and is crucial for viability of
cells. It is the only lipid not dedicated to metabolism of energy and its storage. The
majority of cholesterol resides in cellular membranes, where it influences the order
of phospholipid chains and contributes to membrane fluidity, integrity and hetero-
geneity. Further roles of cholesterol also include cell cycle regulation and protein
modification, as well as being the starting point for the synthesis of steroid hormones
and bile acids (Fig. 1).
The synthesis of cholesterol is a housekeeping pathway and supposedly takes
place in all mammalian cells. In mammals, there are more than 100 genes associated
with cholesterol synthesis and synthesis regulation [1]. Cholesterol synthesis itself
includes over 20 reactions, starting from acetyl coenzyme A [2]. This chapter
focuses on the second part of cholesterol synthesis, from lanosterol on (Fig. 2). It
is called the post-squalene or sometimes post-lanosterol portion of cholesterol
synthesis since lanosterol is the first sterol intermediate in the pathway. Initially,
the post-squalene pathway has been divided into the Bloch and Kandutsch–Russell
branches [3]. In the Bloch branch, the final reaction is the conversion of desmosterol
to cholesterol by sterol-Δ24-reductase (DHCR24); thus, all intermediates from
lanosterol to desmosterol contain Δ24 double bonds. In contrast, in the Kandutsch–
Russell branch, DHCR24 acts already on lanosterol; thus all intermediates from
24,25-dihydrolanosterol to 7-dehydrocholesterol contain a saturated side chain.
Since DHCR24 can, in principle, metabolize any cholesterol synthesis intermediate
from lanosterol on, the two branches cannot be treated separately. Study of
DHCR24 substrate specificity in vitro showed 24-dehydrolathosterol as the most

C. Skubic · D. Rozman (*)

Faculty of Medicine, Centre for Functional Genomics and Bio-chips, Institute of Biochemistry,
University of Ljubljana, Ljubljana, Slovenia
e-mail: [email protected]

# Springer Nature Switzerland AG 2020 1

D. Rozman, R. Gebhardt (eds.), Mammalian Sterols,
https://doi.org/10.1007/978-3-030-39684-8_1
2 C. Skubic and D. Rozman

Fig. 1 The molecular structure of cholesterol with carbon atoms numbering based on IUPAC-IUB
recommendations. Red ring represents the polar hydroxyl group of cholesterol molecule and the rest
of molecule is nonpolar. The figure represents major roles of cholesterol in the human body. The
major function of cholesterol is in the membrane as the basic part of phospholipid bilayer

reactive substrate, suggesting that cholesterol synthesis preferentially starts with

the Bloch branch from lanosterol to 24-dehydrolathosterol and is then shifted
to Kandutsch–Russell branch via lathosterol. In this case, 7-dehydrocholesterol
presents the last intermediate before cholesterol. According to theoretical
predictions, there could be many more intermediates in cholesterol synthesis [4].
The predicted number of sterol metabolites from lanosterol to cholesterol is 72, com-
prising a metabolic network. This has been calculated by an algorithm that applied a
binary code, labelling each C atom of the sterol ring by either 1, if containing a
substituents, or 0 for no substituents. However, two important facts were not
considered during enzyme activity coding. First, the enzymes SC4MOL, NSDHL,
and HSD17B7 remove the methyl groups at position 4 in a specific order, where
methyl group at position a is removed first. After its removal, methyl group at
position b rotates to position a and is removed in the following step. The coding
system was designed to remove the 4b methyl group first to create molecules that
have one methyl group at position 4a. Second, a single bond at position 7(8) seems
to completely inhibit activity of SC5DL, as no molecules with single bond at 7
(8) and double bond at 5(6) were found in any reports. A single bond at 7(8) may
influence positioning of hydrogen atoms at position 6 and thus may prevent
interaction of substrate with SC5DL [4].
We will now describe the major intermediates of cholesterol synthesis, their roles
as signaling molecules, and links to human diseases.
 Sterols from the Post-Lanosterol Part of Cholesterol Synthesis: Novel. . .

Fig. 2 Cholesterol synthesis with its intermediates. Figure represents the major cholesterol intermediates and their physiological functions in blue circles. The
synthesis is from two parts, the pre-squalene in green square (not discussed in this work) and post-squalene in which lanosterol is the first molecule with sterol
3

structure. With green arrows, the most probable synthesis pathway is shown
4 C. Skubic and D. Rozman

Sterol Intermediates from Lanosterol to Cholesterol

Lanosterol and 24,25-Dihydrolanosterol: The Branch Points of Sterol

Synthesis

Lanosterol is the first cyclic intermediate of cholesterol synthesis pathway and the
first sterol molecule in the post-squalene part of the synthesis (Sterol structures in
Table 1). Lanosterol is formed from squalene by squalene epoxidase/monooxsyenase
(SQLE) and lanosterol synthase (LSS) [5]. From lanosterol on the cholesterol synthe-
sis is done by enzymes that are membrane bound in endoplasmic reticulum [6]. In
the Bloch branch lanosterol is converted into FF-MAS (Follicular fluid meiosis-
activating sterol) by CYP51 enzyme [7].
Addition of lanosterol efficiently stabilizes co-chaperon CHIP. CHIP acts as a
molecular switch for both proteasomal and lysosomal mechanisms, which reduce
aggregation of misfolded proteins, that often cause pathological conditions
[8]. According to some studies, lanosterol surprisingly reverses protein aggregation
in cataracts of animals. Lanosterol can dissolve precipitates and even amyloid-like
fibril structures, which are the cause of cataracts in individuals. Lanosterol effec-
tively treats cataracts in rabbit and dog lenses in vivo. Treating the cataract lenses
with lanosterol was not yet successful in humans [9]. The “dissolving” ability of
lanosterol seems to be partially controversial. Sterol intermediates of cholesterol
synthesis, in particular desmosterol (discussed later), have been accused to cause
cataracts and resulted in removal of a cholesterol lowering drug Triparanol from the
market in early 1960s, as reviewed [10]. Triparanol was supposed to lower the level
of cholesterol in the blood and reduce the risk of heart attacks. However, many
people who took the drug went blind from an unusual form of cataract. It was
suggested that accumulating desmosterol is causing the cataracts, and consequently,
that all sterol intermediates toward cholesterol, from lanosterol and on, likely have
harmful effects and should not accumulate in the cells. This also negatively
influenced further drug development efforts to find novel hypolipidemic drugs
with non-statin properties, that would inhibit enzymes in the late part of cholesterol
synthesis [10].
For lanosterol, it was proven to have toxic effects in CHO-7 cells. Addition of
lanosterol to cholesterol auxotrophs failed to support growth and killed the cells.
Surprisingly, lanosterol killed also the wild-type cells, which underlines its toxic
effect. The explanation has been that lanosterol indirectly suppressed cholesterol
synthesis [11] and promoted the degradation of 3-hydroxy-3-methylglutaryl-coen-
zyme A reductase (HMG CoA reductase), and in this way downregulated cholesterol
synthesis [12]. Alternative explanations claimed that sterols that have two methyl
groups at position 4, such as lanosterol, cannot replace cholesterol in membranes,
which results in defective membranes and cell death.
Recently Hubler et al. discovered new roles of sterols that have a double bond
between C8 and C9 like lanosterol, FF-MAS, T-MAS, and zymostenol. Their results
are showing that these sterols have an important role in oligodendrocytes formation.
Table 1 The 19 known cholesterol intermediates, from lanosterol toward cholesterol
Molecular weight Converted
Trivial name Chemical name Formula Structure (g/mol) by
Lanosterol 4,4,14α-Trimethyl-5α-cholesta-8(9),24- C30H50O 426.72 CYP51A1
dien-3β-ol

24,25-Dihydrolanosterol 4,4,14α-Trimethyl-5α-cholest-8(9)en- C30H52O 428.73 CYP51A1

3β-ol

FF-MAS 4,4-Dimethyl-5α-cholesta-8(9),14,24- C29H46O 410.67 TM7SF2,

trien-3β-ol LBR

Dihydro-FF-MAS 4,4-Dimethyl-5α-cholesta-8(9),14-dien- C29H48O 412.69 TM7SF2,

3β-ol LBR
Sterols from the Post-Lanosterol Part of Cholesterol Synthesis: Novel. . .

T-MAS 4,4-Dimethy-5α-cholesta-8(9),24-dien- C29H48O 412.69 SC4MOL

3β-ol
5

(continued)
 6
Table 1 (continued)
Molecular weight Converted
Trivial name Chemical name Formula Structure (g/mol) by
Dihydro-T-MAS 4,4-Dimethy-5α-cholest-8(9)-en-3β-ol C29H50O 414.69 SC4MOL

4-Methyl-4- C29H46O3 442.34 NSDHL

carboxyzymosterone

4-Methyl-4-carboxy C29H48O3 444.36 NSDHL

zymostenone

4-Methyl C28H44O 396.34 HSD17B7

zymosterone

4-Methyl C28H46O 398.35 HSD17B7

zymostenone
C. Skubic and D. Rozman
4-Methyl C28H46O 398.35 SC4MOL
zymosterol NSDHL
HSD17B7

4-Methyl C28H48O 400.37 SC4MOL

zymostenol NSDHL
HSD17B7

Zymosterol 5α-Cholesta-8(9),24-dien-3β-ol C27H44O 384.64 EBP

Zymostenol 5α-Cholest-8(9)-en-3β-ol C27H46O 386.65 EBP

Sterols from the Post-Lanosterol Part of Cholesterol Synthesis: Novel. . .

24-Dehydrolathosterol 5α-Cholesta-7,24-dien-3β-ol C27H44O 384.64 SC5DL

(continued)
7
 8

Table 1 (continued)
Molecular weight Converted
Trivial name Chemical name Formula Structure (g/mol) by
Lathosterol 5α-Cholest-7-en-3β-ol C27H46O 386.66 SC5DL

7-Dehydrodesmosterol 5α-Cholesta-5,7,24-trien-3β-ol C27H46O 386.66 DHCR7

7-Dehydrocholesterol 5α-Cholesta-5,7-dien-3β-ol C27H44O 384.64 DHCR7

Desmosterol 5α-Cholesta-5,24-dien-3β-ol C27H44O 384.64 DHCR24

Cholesterol 5α-Cholest-5-en-3β-ol C27H46O 386.66

Each sterol is represented with its molecule structure, chemical formula, molecular weight, and enzyme uses this sterol as substrate in the cholesterol synthesis
pathway
C. Skubic and D. Rozman
Sterols from the Post-Lanosterol Part of Cholesterol Synthesis: Novel. . . 9

By inhibition of enzymes in sterol synthesis, they found that 8,9-unsaturated sterols

that accumulate in these cells are responsible for the promotion of oligodendrocyte
formation and remyelination. The mechanism through which sterols promote oligo-
dendrocyte formation is poorly understood. It is suspected that nuclear hormone
receptors (NHR) have a role in this, but reporter assays with 20 different NHRs that
are known to interact with sterols have not shown any activity in the case of
oligodendrocytes. This means that the mechanism by which 8,9-unsaturated sterols
promote oligodendrocytes is not yet fully understood [13].
There is no nuclear receptor identified that would be specific for lanosterol. It was
shown that lanosterol can activate RORC but with a lower activity compared to some
other sterols discussed below [14].
24,25-Dihydrolanosterol (DHL) can be converted from lanosterol by enzyme
DHCR24 at the beginning of Kandutsch–Russell pathway of cholesterol synthesis.
DHCR24 catalyzes the reduction of Δ24 bond in lanosterol [2]. But in most tissues
lanosterol is predominantly converted to FF-MAS as proposed in the Bloch path-
way. There is some evidence that 5–10% of lanosterol in the liver is converted to
DHL, but not to downstream intermediates via CYP51 as proposed in Kandutsch–
Russell pathway. There is possibility that some of hepatic DHL is converted to
another sterol, the identity of which remains unknown [15].
DHL has a major role in posttranscriptional regulation of HMGCR (3-hydroxy-3-
methylglutaryl-coenzyme A reductase), which catalyzes the reduction of HMG CoA
(3-hydroxy-3-methylglutaryl-coenzyme A) to mevalonate. This is a rate-limiting
step in the entire cholesterol synthesis. DHL inhibits HMGCR by triggering degra-
dation and ubiquitination of the enzyme. DHL causes conformational changes in
HMGCR to enable interactions with the INSIG proteins. INSIG proteins recruit the
E3 ubiquitin ligase gp78 and proteasome-associated protein VCP, which results in
ubiquitination and degradation of HMGCR. In this way, DHL affects cholesterol
synthesis and its homeostasis [16]. Because DHL is supposedly converted from
lanosterol mainly in the liver, the function of cholesterol regulation by DHL through
HMGCR is likely valid only for the liver [15].

Meiosis Activating Sterols FF-MAS and T-MAS

Follicular fluid meiosis-activating sterol (FF-MAS) was first identified in human

ovarian follicular fluid [17]. It is a cholesterol synthesis intermediate from the Bloch
pathway. It is converted from lanosterol by the previously mentioned enzyme
lanosterol 14α-demethylase (CYP51). CYP51 belongs to the cytochrome P450
superfamily and converts lanosterol (and DHL) into FF-MAS together with micro-
somal electron transferring protein NADPH-P450 reductase [18]. The highest
concentrations of FF-MAS are found in the ovary, for instance in human preovula-
tory follicular fluid the concentration of FF-MAS is around 1.3 μM (T-MAS
concentration is half of FF-MAS) [19]. The next step in cholesterol synthesis is
reduction of Δ14 bond in FF-MAS to form testis meiosis-activating sterol (T-MAS)
that will be discussed in detail in the next paragraph. This reduction can be done by
10 C. Skubic and D. Rozman

two different enzymes encoded by two different genes, the sterol-Δ14-reductase

(DHCR14) and lamin B receptor (LBR) [2]. There is evidence suggesting that mice
models with DHCR14 deletion were still able to normally synthesize cholesterol
[20]. On contrary, HeLa cell lines with LBR knockout were unable to effectively
sustain cholesterol synthesis, despite the presence of DHCR14, which means LBR is
necessary for normal cholesterol synthesis [21]. FF-MAS can also be converted by
enzyme DHCR24 to form FF-MAS with saturated side chain—dihydro-FF-MAS,
which is a part of Kandutsch–Russell pathway [2].
Meiosis activating sterols (MAS) were first isolated from preovulatory follicular
fluid from women undergoing treatment for infertility by in vitro fertilization
(FF-MAS) and from bull testicular tissue (T-MAS). MAS function as stated in
name, was first associated with function of activation of oocyte meiosis [17].
Meiosis is crucial for sexual reproduction of animals and forming haploid geneti-
cally balances gametes. In mammals, oogonia enter meiosis and are transformed into
oocytes. Oocytes are arrested in late prophase of the first meiotic division at diploid
stage. Meiosis does not resume until the follicular unit is not mature, fully grown,
and can respond to gonadotropin hormones (FSH, LH), which resume meiosis and
ovulation [19]. FF-MAS has the ability to resume meiosis of oocyte [17] in the
gonadotropin-dependent mechanism. FF-MAS probably works through receptor-
mediated mechanism and efforts have been invested to find specific MAS receptor.
Liver X receptor (LXR) is a nuclear receptor that binds FF-MAS and several
oxysterols. It was a candidate for receptor-mediated mechanism of oocyte meiosis
resumption, but was excluded because FF-MAS has lower affinity for LXR than
oxysterols [22]. Another reason is that none of oxysterols that can bind to LXR was
able to resume oocyte meiosis [19]. Data indicate that MAS signaling may work
through a G-protein-coupled mechanism and may be mediated by binding to an
oocyte plasma membrane-associated protein with high affinity for FF-MAS. There is
a lot of evidence suggesting the role of FF-MAS in resumption of oocyte meiosis,
but another set of data suggested that FF-MAS might not be a universal mediator of
hormone-induced meiotic maturation. In vivo role of MAS is not clear and stays
enigma [23], some experimental evidence oppose the in vivo roles of MAS. The
delay in germinal vesicle breakdown after addition of MAS or AY-9944, an inhibitor
of DHCR7, a later enzyme of cholesterol synthesis, was at that time the strongest
evidence against the suggested role of MAS as an essential mediator of luteinizing
hormone in meiosis resumption [24]. It was, on the other side, proposed that the
MAPK signaling pathway is required for the MAS-like resumption of meiosis
activated through one of the hormonal upstream pathways [25].
T-MAS is converted from FF-MAS with enzymes DHCR14 and LBR [2]. It was
first isolated from bull testicular tissue using high-performance liquid chromatog-
raphy [17]. T-MAS is the part of cholesterol synthesis pathway and is generated
in all cholesterol-synthesizing cells. In most cell types T-MAS immediately
converted to zymosterol [26] through several intermediates catalyzed by sterol
4,4-dimethylase enzyme complex. The first step is carboxylation of one of the
4-methyl groups by enzyme sterol-C4-methyl oxidase (SC4MOL) to form
4-methyl-4-carboxyzymosterone. 3β-hydroxy-Δ5-steroid-dehydrogenase (NSDHL)
Sterols from the Post-Lanosterol Part of Cholesterol Synthesis: Novel. . . 11

then converts 3β-hydroxy group to 3-keto by removing one of the CO2 and
4-methylzymosterone is formed, which is then converted to 4-methylzymosterol by
enzyme 3β-keto-reductase (HSD17B7) by restoring 3β-hydroxy group. Enzymatic
process is then repeated one more time and zymosterol is formed [2]. FF-MAS and
T-MAS both accumulate at high concentrations in ovaries and testes, due to low
expression of respective downstream enzymes [26]. In mammalian testes T-MAS
concentration can be above 30 μg/g, FF-MAS in testes is present only in trace
amounts [19]. The large fraction of T-MAS that is synthesized in testes is not
converted to zymosterol, but to sterol that remains unidentified and is not a part of
the downstream cholesterol synthesis [15]. The elevated production and accumula-
tion of T-MAS in the testis may result from transcriptional regulation of cholesterol
synthesis pathway and inhibition of the enzymes converting T-MAS into downstream
cholesterol synthesis intermediates. CYP51 can escape the SREBP regulation under
certain physiological conditions in the testes and can be upregulated via cAMP-
dependent stimuli and cAMP-responsive element modulator (CREMτ). CREMτ is
expressed only in spermatids and regulates genes associated with maturation and
development of spermatids [23].
It is still not firmly established whether T-MAS is as FF-MAS associated with the
resumption of meiosis. In vitro T-MAS and FF-MAS have interchangeable roles and
are able to induce meiosis resumption in similar concentrations. T-MAS is more
efficient in germinal vesicle breakdown assay in naked oocyte [23]. T-MAS concen-
tration in testes is high, but its function is not clear. It may play a role in fertilization,
spermatogenesis, and resumption of second meiotic division [19]. Since CYP51 that
converts lanosterol to FF-MAS, unequivocally localizes to acrosomal membranes of
male germ cells, it was proposed that sperm cells can synthesize MAS sterols in situ
[27]. To test the role of sterols MAS in vivo, we generated a conditional male germ
cell-specific knockout of Cyp51 in the mouse. As expected, metabolic profiling
revealed elevated CYP51 substrates lanosterol and 24,25-dihydrolanosterol and
diminished levels of MAS, the immediate products of CYP51. To our surprise the
germ cell-specific ablation of Cyp51 did not affect testicular morphology, sperm
production, or reproductive performance of the males. These results failed to show
that de novo synthesis of MAS and cholesterol in male germ cells is most likely not
essential for spermatogenesis [28].

Zymosterol and Zymostenol

Zymosterol is synthesized from T-MAS with removal of two methyl groups on C4 in

the process involving enzymes SC4MOL, NSDHL, and HSD17B7. In the Bloch
pathway zymosterol is then converted to 24-dehydrolathosterol by enzyme sterol-
Δ8–7-isometase (EBP), which shifts the double bond from position Δ8 to position Δ7.
Like all cholesterol intermediates in the Bloch pathway, zymosterol can also be
theoretically converted by DHCR24 to zymostenol in K–R pathway [2, 29]. But in
the case of zymosterol, there is evidence that this may actually be one of the points of
crossover between Bloch and the Kandutsch–Russell pathway. This was confirmed
12 C. Skubic and D. Rozman

on embryotic kidney cells HEK-293. Some data show that zymosterol is a better
substrate for DHCR24 than lanosterol, which is the first intermediate in the Bloch
pathway, which means that crossover between both pathways is probably down-
stream of lanosterol. Anyway, in most cells zymosterol is not considered to be the
main substrate for DHCR24; its major physiological substrate is desmosterol on the
end of Bloch pathway [15].
Zymosterol is synthesized like other sterols in rough ER. Some of the newly
synthesized zymosterol in cultured human fibroblasts is rapidly (even faster than
cholesterol) transferred to the plasma membrane. Then some of zymosterol is
transferred back to ER, where it is converted to next sterol intermediates and finally
to cholesterol. The function of this movement within the cell is unknown [30]. In
studying zymosterol effect on artificial lipid monolayer, it was concluded that
zymosterol possesses condensing and ordering abilities. However, the effect of
zymosterol on lipid membranes is much less efficient compared to cholesterol [31].
Zymosterol is considered one of the ligands for the nuclear receptor RORγ
(discussed in last chapter). Altered zymosterol concentration can through binding
with RORγ [32] change the expression of genes associated with immunity, circadian
rhythm, and metabolism [33]. Like other 8,9-unsaturated sterols zymosterol can also
enhance the oligodendrocyte formation [13].
Zymostenol can be synthesized from zymosterol with enzyme DHCR24 [2] and
is also one of the crossover points between Bloch and Kandutsch–Russell pathway
of cholesterol synthesis in some tissues [15]. Zymostenol can theoretically be
synthesized also from dihydro-T-MAS, an upstream cholesterol intermediate in
Kandutsch–Russell pathway, through series of reactions with enzymes SC4MOL,
NSDHL, and HSD17B7, like in T-MAS to zymosterol conversion [2], but this
pathway is not supported with sufficient evidence. Sterols proceed down the Bloch
pathway at least until zymosterol, when the demethylation of sterol nucleus is
complete, and can then be converted by DHCR24 and cross into Kandutsch–Russell
pathway [15].

Lathosterol and 24-Dehydrolathosterol

Lathosterol is the next cholesterol intermediate from Kandutsch–Russell pathway

downstream of zymostenol. It is converted from zymostenol by enzyme EBP or from
24-dehydrolathosterol with enzyme DHCR24 [2, 29]. There is more evidence that
physiologically lathosterol is synthesized from zymostenol and not from
24-dehydrolathosterol [15]. In the next step of cholesterol synthesis, lathosterol is
converted to 7-dehydrocholesterol by enzyme SC5DL, which catalyzes the forma-
tion of double bond between C5 and C6.
24-Dehydrolathosterol is converted from zymosterol with changing double bond
position form Δ8 position to Δ7 with enzyme EBP. In Bloch pathway enzyme sterol-
C5-desaturase/lathosterol oxidase (SC5DL) formats double bond on C5 position and
converts 24-dehydrolathosterol to 7-dehydrodesmosterol. 24-Dehydrolathosterol
can also be converted by DHCR24 to lathosterol which is intermediate from
Kandutsch–Russell pathway [2].
Sterols from the Post-Lanosterol Part of Cholesterol Synthesis: Novel. . . 13

7-Dehydrocholesterol

7-Dehydrocholesterol (7-DHC) is the last intermediate in Kandutsch–Russell cho-

lesterol synthesis pathway and it is converted from lathosterol by enzyme SC5DL.
The last reaction before cholesterol is synthesized is reduction of Δ7 double on
7-deydrocholesterol by enzyme DHCR7 [2, 34].
Apart from being precursor for cholesterol, 7-DHC is also precursor for vitamin D
synthesis. Vitamin D3 is produced in a two-step nonenzymatic process. UVB light
(280–320 nm wavelength) brakes the B-sterol ring and pre-vitamin D3 is formed,
which then isomerizes to form vitamin D3. This process happens in the skin and is
dependent on UVB intensity and melanin (skin pigmentation level), which can block
UVB from reaching 7-dehydrocholesterol [35].
It was shown that 7DHC can destabilize the HMGCR enzyme. In this way 7DHC
can downregulate cholesterol production. This function of 7DHC was shown only in
the case of 7DHC accumulation like in case of SLOS (discussed in next chapter)
patients [36].

Desmosterol and 7-Dehydrodesmosterol

Desmosterol is synthesized from 7-dehydrodesmosterol by enzyme DHCR7, which

removes double bond from Δ7 position. It is the last intermediate in Bloch choles-
terol synthesis pathway and is converted to cholesterol by enzyme DHCR24, which
catalyzes the reduction of Δ24 double bond on the side chain of sterol ring [34].
Desmosterol also binds to LXR, which controls cholesterol export genes and
represses inflammatory genes. Desmosterol can accumulate in macrophage foam
cells and by LXR activation regulates metabolism and inflammatory response
[29]. In mature testis, desmosterol can be found in high concentration (up to 12%
of total sterols) and even higher in sperm where desmosterol can reach concentration
up to 58% of total sterols (both data are for Rhesus monkeys). The high desmosterol
level may be the consequence of lower levels of DHCR24 or its inhibition. In sperm,
concentration of desmosterol is not homogenous, but desmosterol is mainly located
in sperm tail. This effects the membrane fluidity and is possible to be necessary for
normal motility of flagella [37].
Experiments done on J774 cells (mouse monocyte/macrophage) showed that cells
substitute cholesterol with desmosterol in case of cholesterol depletion. Cells were
able to survive and proliferate without cholesterol (DHCR24 mutation and medium
without cholesterol). Because desmosterol and cholesterol differ only in double bond
on position C24, and it is not surprising that desmosterol can substitute cholesterol in
some of the processes. J774 cells with cholesterol depletion have a functional
SREBP pathway and desmosterol can suppress SREBP pathway and gene expres-
sion [38]. Similarly to zymosterol, desmosterol also binds and activates the RORγ
and affects the expression of genes controlled by this nuclear receptor [32].
In the Bloch pathway 7-dehydrodesmosterol is converted from
24-dehydrolathosterol by the enzyme SC5DL, which catalyzes the formation of