Placenta xxx (2017) 1e8

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Placenta
journal homepage: www.elsevier.com/locate/placenta

Notch signalling in placental development and gestational diseases

er*
S. Haider, J. Pollheimer, M. Kno
Department of Obstetrics and Gynaecology, Reproductive Biology Unit, Medical University of Vienna, Vienna, Austria

a r t i c l e i n f o a b s t r a c t

Article history: Activation of Notch signalling upon cell-cell contact of neighbouring cells controls a plethora of cellular
Received 13 December 2016 processes such as stem cell maintenance, cell lineage determination, cell proliferation, and survival.
Received in revised form Accumulating evidence suggests that the pathway also critically regulates these events during placental
9 January 2017
development and differentiation. Herein, we summarize our present knowledge about Notch signalling
Accepted 12 January 2017
in murine and human placentation and discuss its potential role in the pathophysiology of gestational
disorders. Studies in mice suggest that Notch controls trophectoderm formation, decidualization,
Keywords:
placental branching morphogenesis and endovascular trophoblast invasion. In humans, the particular
Placenta
Trophoblast
signalling cascade promotes formation of the extravillous trophoblast lineage and regulates trophoblast
Notch signalling proliferation, survival and differentiation. Expression patterns as well as functional analyses indicate
Gestational diseases distinct roles of Notch receptors in different trophoblast subtypes. Altered effects of Notch signalling
have been detected in choriocarcinoma cells, consistent with its role in cancer development and pro-
gression. Moreover, deregulation of Notch signalling components were observed in pregnancy disorders
such as preeclampsia and fetal growth restriction. In summary, Notch plays fundamental roles in
different developmental processes of the placenta. Abnormal signalling through this pathway could
contribute to the pathogenesis of gestational diseases with aberrant placentation and trophoblast
function.
2017 Elsevier Ltd. All rights reserved.

1. General role of Notch signalling
Notch represents an evolutionarily conserved signalling
pathway critically involved in embryonic development and tissue
homeostasis of adult organs. It controls a vast range of cellular
processes including proliferation, survival, differentiation and
motility. Signalling through Notch requires direct cell-cell contact
allowing for short-range communication between neighbouring
cells [1]. Hence, the pathway is ideally suited to control self-
renewal and differentiation in stem cell niches [2,3]. In the latter,
Notch was shown to mediate binary cell fate decisions: different
levels of Notch activation are created in two initially equipotent,
adjacent precursor cells thereby promoting adoption of distinct cell
fates. For example, in the developing mouse brain Notch inhibited
neuronal differentiation, but induced a glial cell fate and promoted
astrocyte differentiation [2]. Likewise, Notch signalling is crucial for
epithelial stemness and cell fate specication. It maintains murine
* Corresponding author. Medical University of Vienna, Department of Obstetrics
and Gynaecology, Reproductive Biology Unit, Waehringer Guertel 18-20, A-1090
Vienna, Austria.
E-mail address: martin.knoeer@meduniwien.ac.at (M. Kno er).
intestinal stem cells and balances the cell fate of terminally
differentiating gut cells [3]. Interestingly, Notch was shown to have
an opposed role in intestinal stem cells of Drosophila, impairing
their self-renewal. Hence, Notch signalling is highly complex and
depends on the cellular context as well as on its cross-talk with
other developmental signalling cascades mediated through mem-
bers of the Wingless (Wnt), epidermal growth factor (EGF) or
transforming growth factor-b (TGF-b) family [4,5]. Conditions of the
particular microenvironment such as composition of the extracel-
lular matrix, shear stress or hypoxia also determine the various
effects of Notch in cells and tissues [4]. Accordingly, Notch signal-
ling components may act as tumour suppressors or oncogenes in
different organs and differentially inuence cancer development
and progression [6e8]. Considering its regulatory role in de novo
vessel formation, angiogenesis and branching morphogenesis
[9e11], it may not be surprising that Notch also controls these
processes during placental development [12]. The present review
summarises our current knowledge about Notch signalling in mu-
rine and human placentation and discusses its role in the context of
gestational diseases and choriocarcinoma cell function.

http://dx.doi.org/10.1016/j.placenta.2017.01.117
0143-4004/ 2017 Elsevier Ltd. All rights reserved.

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1.1. Canonical Notch signalling
Canonical Notch signalling is activated upon interaction of a
membrane-anchored ligand on the signal-sending cell with a Notch
receptor on the adjacent signal-receiving cell [1,13]. In contrast,
binding of the ligand to a receptor on the same cell was shown to
inhibit the pathway [14]. In mice and humans four different Notch
receptors (Notch1-4) and ve distinct Notch ligands, the Serrate-
like ligands (Jagged1 and 2) and the Delta-like ligands (DLL1, 3
and 4) have been identied [1]. Notch receptors are single trans-
membrane proteins. After translation the Notch polypeptide chain
is cleaved by furin-like convertases generating a heterodimer of the
Notch extracellular domain (NECD) non-covalently linked to the
Notch transmembrane and intracellular domain (NTM-ICD) (Fig. 1).
NECD harbours epidermal growth factor (EGF)-like repeats neces-
sary for ligand interaction. These repeats can be modied by
different types of glycosylation thereby governing differential
ligand binding and receptor activation [15].
NTM-ICD contains a trans-membrane domain (TMD) and
different intracellular protein motifs required for the nuclear
function of the Notch intracellular domain (NICD). The latter arises
after two sequential cleavages from the receptor [1]. Upon inter-
action of Notch with its ligand, members of the ADAM family, such
as ADAM10 or ADAM17, cleave NTM-ICD 12 amino acids before the
trans-membrane domain thereby generating the intermediary
membrane protein Notch extracellular truncation (NEXT). Subse-
quently, NEXT is recognized by g-secretase [16], cutting the protein
at two different cleavage sites in the TMD. These proteolytic steps
release NICD from the membrane allowing it to translocate into the
nucleus where it acts as a co-activator of transcription. Besides a
transactivation domain, NICD comprises a RAM (RBPJk-associated
module) domain for high afnity binding of the transcription factor
recombination signal binding protein for immunoglobulin kappa J
(RBPJk) as well as ankyrin repeats for RBPJk activation through
binding of co-activators of the mastermind-like (MAML) family. In
the absence of NICD RBPJk operates as a repressor recruiting his-
tone deacetylases and other co-repressors such as CtBP (C-termi-
nal-binding protein 1), CIR (CBF1 interacting co-repressor) or
SHARP (SMRT and HDAC associated repressor protein) [5]. Upon
Notch activation these repressors are displaced by NICD thereby
activating Notch target genes after binding of MAML proteins and
further co-activators [17]. Activity of canonical Notch signalling is
regulated by multiple mechanisms controlling post-translational
Notch modication, receptor availability at the membrane, and
expression of soluble Notch ligands, discussed elsewhere [1,13].
The expression of Notch targets strictly depends on the cell- and
tissue-specic role of the pathway. Whereas NICD induces prolif-
eration by directly activating myc, cyclin D1 and CDK5, it promotes
growth arrest and differentiation by inducing kinase inhibitors
such as p21 [18e20]. However, the prime targets of canonical Notch
signalling are members of the Hairy/Enhancer of Split (HES) family,
HES1, HES5, HES7 and Hairy/Enhancer-of-split related with YRPW
motif (HEY) proteins, HEY1, HEY2 and HEYL [21]. HES and HEY
proteins are basic helix loop helix proteins (bHLH) acting as

Fig. 1. The canonical Notch pathway. Membrane-anchored Notch ligands contain a transmembrane domain (TDM), a cysteine (C)-rich domain and EGF-like repeats for binding to
Notch receptors. Moreover, the N-terminal DSL (Delta/Serrate/LAG-2 proteins) domain and the DOS (Delta and OSM-11-like proteins) motif contribute to receptor binding. In
contrast to Jagged 1 and Jagged 2, the C-rich region is absent from DLL1, whereas DLL3 and DLL4 additionally lack the DOS domain [1]. The co-ligands Delta-like homologue 1 and 2
(DLK1 and 2) harbour a DOS domain but do not contain a DSL motif. Depending on the receptor subtype, the Notch extracellular domain (NECD) contains differential numbers of
EGF-like repeats. In addition, NECD comprises cysteine-rich Lin12-Notch repeats (LNR) and a heterodimerization domain (HD) for binding to the Notch transmembrane and
intracellular domain (NTM-ICD). Upon Notch activation, cleavage by ADAM proteases generates the Notch extracellular truncation (NEXT) which is further processed by g-secretase
yielding Notch intracellular domain (NICD). The latter harbours a nuclear localisation sequence (NLS), the RBPJk-associated module (RAM) for binding to RBPJk and ankyrin (ANK)
repeats for interaction with mastermind-like (MAML) proteins. Nuclear translocation of NICD displaces co-repressors (Co-R) from RBPJk and recruits co-activators (Co-A) thereby
promoting transcription of canonical Notch target genes such as repressors of the HES and HEY family. Stability of NICD is regulated via ubiquitination of proline/glutamic acid/
serine/threonine-rich (PEST) motifs and proteosomal degradation.

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transcriptional repressors, particularly upon binding to co-
repressors of the Groucho/Transducin-like enhancer of split (TLE)
family [22,23]. These protein complexes are critically involved in
the maintenance of stem and progenitor cells by repressing key
regulators of cell fate determination.
Along those lines, the role of canonical Notch signalling in
stemness and cell line lineage decisions is highly complex and
explained by different regulatory mechanisms [2,3,13]. In lateral
inhibition equivalent precursors undergo different cell fates since
high expression of a Notch ligand in a given cell activates the
pathway in a neighbouring cell. In the latter, NICD-RBPJk prevents
Notch ligand expression through the induction of HES. As a
consequence of subsequent feedback loops, the ligand-expressing
cell is maintained in a Notch-inactive state whereas the activated
cell adopts a different cell fate. Alternatively, Notch-dependent cell
fate decisions are regulated by inhibitors of the NUMB family which
have been described as multifunctional adaptor proteins involved
in endocytosis and intracellular trafcking of membrane-bound
molecules [24]. NUMBs interact with proteins of the endocytic
machinery, such as a-adaptin and Eps15, thereby promoting
internalisation and sequestration of Notch receptors. Moreover,
NUMB was shown to antagonise canonical Notch signalling by
binding to NTM-ICD and membrane-tethered NICD, thus recruiting
E3-ubiquitin ligases for proteasomal degradation. The adoption of
distinct cell fates is achieved by asymmetrical segregation of NUMB
during cell division of a progenitor, thereby creating two different
daughter cells with active and inactive Notch signalling, respec-
tively [13].
1.2. Non-canonical Notch signalling
Besides the canonical activation of RBPJk, full-length Notch re-
ceptors as well as NICD exert non-canonical effects in cells and
tissues in a ligand-dependent or -independent manner [25]. Notch
has been initially identied as a cell adhesion molecule in
Drosophila and was also shown to promote cell-cell interactions in
mammals independently of NICD generation [26]. Moreover, NTM-
ICD and/or membrane-tethered NICD bind signalling components,
such as phosphatidylinositol-3-kinase (PI3K) and mammalian
target of rapamycin C2 (mTORC2) [27], promoting cell survival via
AKT (Fig. 2.) Notch-mediated activation of these kinases suppresses
pro-apoptotic p53 or stabilizes X-linked inhibitor of apoptosis
(XIAP) [28,29]. Moreover, nuclear NICD interacts with a variety of
non-canonical transcription factors, for example nuclear factor kB
(NFkB), hypoxia-inducible factor 1a (HIF1a) or Ying Yang1 (YY1)
[30e32], promoting their nuclear retention and/or activation of
Notch targets upon binding to RBPJk (Fig. 2). Notch enhances NFkB
signalling at various levels [33]. In addition to direct binding of the
NFkB p52 subunit, NICD interacts with IkB kinase a (IKKa) pro-
moting phosphorylation of inhibitor of kB (IkB). Degradation of the
latter allows for nuclear translocation of the active transcription
factor and expression of its target genes. Notch's cross-talk to Wnt
signalling is mediated through binding of its central player, active
b-catenin, to the ankyrin repeats of the full-length receptor. Sub-
sequently, Notch promotes lysosomal degradation of active b-cat-
enin involving NUMB-mediated endocytosis thereby antagonizing
canonical Wnt signalling [34].
2. Notch signalling in reproductive tissues
Besides other pathways Notch-mediated cell-cell communica-
tion plays a crucial role in the development of reproductive organs
and their homeostasis in adults. In females, the particular signalling
cascade controls formation of the reproductive tract, folliculo-
genesis, proliferation of granulosa cells during oogenesis, as well as
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3
oocyte survival and maturation [35e37]. Likewise, Notch is also
involved in spermatogenesis [38]. Abnormal Notch activation has
been noticed in ovarian and testicular cancer [39,40]. Functions of
individual Notch proteins and their cross-talk with other signalling
pathways in germ cells are above the scope of this review and
discussed elsewhere [41].
2.1. Notch signalling in blastocyst formation, implantation and
decidualization
Expression of Notch signalling components has been observed
in embryos commencing with the 4 cell-stage suggesting that the
pathway could be required for pre-implantation development [42].
Moreover, Notch expression in murine and human trophectoderm
(TE) suggested that it could be involved in early lineage decisions
[43,44]. However, homozygous mutation of RBPJk in mice did not
affect progression of zygotes to the blastocyst stage indicating that
Notch signalling might not be necessary for early embryonic
development [45]. On the other hand, RBPJk was shown to also
have Notch-independent roles, largely by acting as a transcriptional
repressor [46]. Indeed, a more recent study suggested that Notch1
ICD promotes expression of the key regulator of TE formation,
caudal-related homeobox transcription factor 2 (Cdx2), in
conjunction with the Hippo signalling-dependent activator TEAD4
and converts outer blastomere cells to a TE cell fate [47].
Notch signalling also critically regulates implantation and
decidualization [48]. Notch receptors and ligands are differentially
expressed during the menstrual cycle and have been detected in
the luminal and glandular epithelium of the non-pregnant uterus
as well as in stromal and endothelial cells [48]. Notch4 is markedly
expressed during the proliferative phase, whereas Jagged1, DLL4
and Notch1 are predominantly produced during the mid-secretory
phase [49e52]. Changes in the expression of Notch1, DLL1 and
Jagged1 were noticed in infertile women suggesting that levels of
some of the Notch receptors and ligands are critical for uterine
receptivity [53,54].
Indeed, precisely regulated levels of activated Notch1 are pivotal
for successful implantation and decidualization [55]. Human
chorion gonadotrophin (hCG)-mediated induction of Notch1 was
shown to promote survival of uterine stromal cells and initiated
decidualization, whereas downregulation of Notch1 and up-
regulation of NUMB was required for completion of differentia-
tion [56,57]. Women suffering from endometriosis have decreased
levels of Notch1, Notch4, Jagged2 and DLL4 [58]. Moreover, over-
expression of Notch1 ICD in the murine uterus causes infertility due
to the lack of uterine glands and DNA-methyltransferase 3b-
mediated hypermethylation of the progesterone receptor gene [59].
During the rst trimester of pregnancy Notch2 was the predomi-
nant receptor in decidual stromal cells which additionally express
Jagged1, DLL1 and DLL4, whereas decidual glands produce all Notch
receptors and ligands [60]. Disruption of Notch2 impaired expres-
sion of the differentiation markers prolactin (PRL) and insulin-like
growth factor binding protein 1 (IGFBP1) suggesting that the re-
ceptor is crucial for decidualization [60]. Moreover, different re-
ceptors and ligands were detected on uterine NK (uNK) cells and
DLL1 and DLL4 increased interferon-g secretion [60,61].
2.2. Notch signalling components control murine placental
development
Regulation of angiogenesis represents one of the best charac-
terised functions of Notch signalling. DLL4-Notch determines the
cell fate of endothelial precursors towards an arterial or venous
phenotype and controls specication of endothelial tips and stalk
cells as well as vessel stability and sprouting [11]. Likewise,
4 S. Haider et al. / Placenta xxx (2017) 1e8

Fig. 2. Non-canonical effects of Notch. Full-length Notch receptor, NTM-ICD and NICD provide a binding interface for multiple proteins exhibiting transcriptional or kinase activity.
Notch increases cell survival by activating PI3K/AKT/mTOR signalling and negatively affects Wnt-b-catenin signalling by triggering degradation of active b-catenin. Nuclear NICD
interacts with different transcription factors such as HIF1a or YY1 in a complex with RBPJk, bound to its cognate sequence, thereby integrating different environmental signals.
Furthermore, Notch promotes activation of NFkB through direct binding of its p52 subunit and indirectly by promoting phosphorylation and proteosomal degradation of IkB through
activation of IKKa.

homozygous mutations of Notch and its target genes mainly affect
the murine placental vascular system and/or branching morpho-
genesis of chorionic villi which goes in hand with vessel sprouting
[12]. Disruption of Notch1, Notch1/Notch4, Dll4, Hey1, Hey2 or
RBPJk inhibited chorio-allantoic branching and/or formation of an
appropriate labyrinthine vascular network around midgestation
[62e67]. In addition, failures in chorion-allantoic fusion and
decreased numbers of proliferative trophoblasts have been re-
ported in RBPJk mutant mice [62,68].
Like in invasive human EVTs [69], Notch2 seems to be the pre-
dominant receptor in diverse mouse trophoblast subtypes. Notch2
has been detected in the chorion, the ectoplacental cone, in spon-
giotrophoblasts as wells as in sinusoidal trophoblast giant cells
(TGCs) and TGCs lining the maternal blood vessels [70e72]. Com-
plementary expression of Notch2 and its putative ligands DLL1,
DLL4, Jagged1 or Jagged2 has been detected in adjacent cell layers
of the chorion, junctional zone and labyrinth, respectively, sug-
gesting that the receptor could orchestrate different steps of
placental development [73]. Although the receptor may not play a
role in trophoblast cell type specication, it is critically involved in
establishing the placental-maternal circulation. Notch2 gene
knock-out prevented formation of blood sinuses and caused
delayed placental perfusion [70]. The role of Notch2 in murine
placentation was investigated in more detail using a homozygous
deletion of Notch2 in spongiotrophoblast precursors giving rise to
invasive trophoblast subtypes, spiral artery-associated TGCs and
canal TGCs [74]. Conditional deletion of the gene inhibited arterial
invasion and reduced the size of maternal blood canals as well as
perfusion indicating a crucial role for Notch2 in trophoblast-
mediated vessel remodelling and differentiation of the above
mentioned TGC subtypes [72]. A similar phenotype was observed in
TLE3 mutant mice suggesting that Notch2 and TLE3 operate in the
same pathway [75].
2.3. Notch signalling regulates human placental development
Notch receptors and their ligands display trophoblast-subtype
specic expression in the human placenta and also vary with
gestational age and trophoblast differentiation [72,76,77]. Although
not functionally tested, expression of Notch1, DLL1, DLL4 and Jag-
ged1 in endothelial cells of villi suggests a role in placental vessel
formation and/or angiogenesis [77]. Akin to its expression in
invasive mouse trophoblasts, Notch2 was predominantly detected
in different extravillous trophoblast (EVT) subtypes of rst and
second trimester tissues and increased during EVT formation
in vivo and in vitro [69,72,76,77]. Interestingly, in distal HLA-G cell
column trophoblasts (CCTs) Notch2 was mainly detected at the cell
membrane, whereas intramural EVTs of remodelling spiral arteries
additionally expressed nuclear Notch2 ICD [69]. Hence, it is antic-
ipated that Notch2-dependent signalling is activated by adjacent
maternal endothelial cells expressing DLL1, DLL4, Jagged1 and
Jagged2 in rst trimester decidual tissues [69]. Human Notch2 ICD
might full a similar role as its murine counterpart being critical for
endovascular trophoblast invasion and adaption of blood ow to
the placenta [72]. In contrast, disruption of Notch2 increased
in vitro motility in human primary trophoblast preparations con-
taining Notch2 distal cell column trophoblasts [69]. Therefore, we
speculate that maintenance of column integrity by cell-cell

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adhesion could be the main function of Notch2 in the distal portion
of the cell column [69].
Similar to other developing systems, the role of Notch signalling
in human placentation is complex and likely depends on the spe-
cic cellular context. Notch1, Notch3 and Notch4 have been
detected in cytotrophoblast (CTB) progenitors whereas Notch2 and
Notch3 are expressed by invasive trophoblasts [72,76]. The overall
Notch activity decreased during in vitro EVT formation suggesting
that the pathway could be mainly associated with maintenance and
stemness of trophoblast precursors [78]. Indeed, Notch1 was
recently shown to be critical for trophoblast survival and prolifer-
ation of villous CTBs and CCT progenitors [24]. Conversely, inhibi-
tion of g-secretase or silencing of RBPJk did not provoke apoptosis
and promoted outgrowth in explant cultures and, as a consequence,
EVT differentiation [24,78]. These contrasting results might be
explained by the Notch-independent roles of g-secretase and RBPJk
as well as the non-canonical functions of Notch1 ICD [79], again
suggesting high complexity of the pathway and individual roles of
its components. RBPJk and total Notch activity could be necessary
to balance rates of cell column growth and EVT differentiation. Of
note, Notch1, regulated by hypoxia, seems to be critical for initia-
tion and maintenance of the EVT lineage [24]. Notch1 appears in
clusters of vCTBs at the basal side of the placenta around the 6th
week of gestation and is specically expressed in a subset of pro-
liferative, proximal CCTs at 12th week. Besides inhibiting cell fusion
and EVT formation, overexpression of Notch1 ICD in CTBs sup-
pressed p63 and TEAD4, two markers of villous CTB self-renewal,
and induced the stemness genes, myc and VE-cadherin, which are
mainly expressed by CCT progenitors. Notch1 exerts its effects by
direct binding to the myc gene as well as by up-regulating IRF6, a
negative regulator of p63 stability [24,80,81]. Rapid down-
regulation of Notch1 during differentiation might involve NUMB
proteins, of which different isoforms have been detected in human
placenta [82].
2.4. Notch signalling in gestational diseases and choriocarcinoma
cells
Gestational diseases such as early-onset preeclampsia and se-
vere intrauterine growth restriction (IUGR) are associated with
defects in trophoblast invasion, spiral artery remodelling and
placental vascularisation [83,84]. Therefore, critical pathways of
placental development such as Notch signalling could be altered in
these pregnancy disorders. Indeed, several studies demonstrated
changes of Notch receptor and ligand expression in preeclamptic
placentae using qPCR, western blotting or immunohistochemistry
[85e89]. However, the majority of these expression data are con-
tradictory which could be explained by differences in placental
sampling or ethnicity, the utilisation of inadequate antibodies,
technical issues in the experimental procedure or differences in
gestational age of placentae. Notch1-4, DLL1, DLL3, DLL4, Jagged1
and Jagged2 were shown to be decreased in preeclamptic tissues
[85,86,88,89], whereas others reported no changes of most Notch
receptors and ligands [72,89], or even an increase of Notch3 [89].
Along those lines, immunohistochemical analyses suggested
Notch1 expression in CTBs and the syncytium of normal,

Fig. 3. The role of Notch in placental anchoring villi. Proximal cell column trophoblasts (CCTs) express Notch1 ICD promoting proliferation and cell survival. Active Notch1 also
induces the extravillous trophoblast (EVT) progenitor-specic stemness markers myc and VE-cadherin and suppresses villous cytotrophoblast (vCTB)-specic TEAD4 and p63
expression, the latter by up-regulating IRF6. Notch1 ICD also suppresses formation of the syncytium (S) and EVT differentiation. One could speculate that Notch1 might also have a
non-canonical role in proximal CCT by binding b-catenin to the membrane. Downregulation of Notch1 in EVTs might then liberate b-catenin from the receptor, thereby provoking its
nuclear recruitment and Wnt-dependent EVT differentiation and/or migration, as recently shown [95]. In contrast to Notch1, Notch2 is present in the distal cell column and in
intramural EVTs. In the latter Notch2-dependent signalling could be required for trophoblast-endothelial interactions promoting endovascular invasion, remodelling and/or short
range communication with immune cells. VS, villous stroma.

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preeclamptic or IUGR placentae obtained between the 24th week
and term [85,87e90]. However, at this time of pregnancy mRNA
and protein of the receptor are completely absent from all
trophoblast subtypes [24,72]. Nevertheless, changes in methylation
of Notch4 and DLL1 were noticed in preeclamptic placentae sug-
gesting epigenetic alterations of Notch components in this disease
[88]. Finally, analyses of Notch signalling in cultivated choriocar-
cinoma cell lines suggested profound changes of its role in tumour
cells. In these studies Notch1 promoted cell migration but inhibited
proliferation [91e93], whereas downregulation of Notch2
increased cell growth [94]. These effects have not been observed
upon manipulation of total Notch activity, Notch1 or Notch2 in
primary CTBs [24,76].
3. Summary
In conclusion, Notch signalling is a critical regulator of murine
and human placental development. In mice, different Notch
pathway members control trophectoderm formation, angiogenesis,
vessel formation, giant cell differentiation and endovascular
trophoblast invasion. In the human placenta, Notch plays a crucial
role in cell column formation, CCT survival and EVT differentiation
(Fig. 3). Despite recent progress much remains to be learned about
Notch in human placentation and trophoblast development. Future
studies should address epigenetic regulation of pathway members
as well as the role of individual Notch ligands and receptors which
have not yet been functionally tested in the human placenta.
Furthermore, the cross-talk of Notch to other developmental
pathways such as Hippo, EGF, TGF-b and Wnt signalling in CTBs
should be elucidated.
Author's contributions
S.H. drew the illustrations and provided support with respect to
literature search and structure of the review. J.P. contributed to
writing and improvement of graphical design. M.K. wrote the
manuscript.
Disclosure statement
The authors have nothing to declare.
Conict of interest
The authors do not report any conict of interest.
Acknowledgements
Research delineating signalling pathways in human trophoblast
development and trophoblast differentiation were supported by
grant P-28417-B30 and P-25187-B13 of the Austrian Science Funds,
Vienna, Austria and by grant 16517 of the Austrian National Bank.
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