Biochimica et Biophysica Acta 1856 (2015) 121–129

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Biochimica et Biophysica Acta

journal homepage: www.elsevier.com/locate/bbacan

Review

The Hippo signal transduction pathway in soft tissue sarcomas

Abdalla D. Mohamed a, Annie M. Tremblay c,d,e, Graeme I. Murray b, Henning Wackerhage a,⁎
a
School of Medical Sciences, University of Aberdeen, AB25 2ZD Scotland, UK
b
School of Medicine and Dentistry, University of Aberdeen, AB25 2ZD Scotland, UK
c
Stem Cell Program, Children's Hospital, Boston, MA 02115, USA
d
Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA
e
Harvard Stem Cell Institute, Cambridge, MA 02138, USA

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

Article history: Sarcomas are rare cancers (≈1% of all solid tumours) usually of mesenchymal origin. Here, we review evidence
Received 10 April 2015 implicating the Hippo pathway in soft tissue sarcomas. Several transgenic mouse models of Hippo pathway
Received in revised form 27 May 2015 members (Nf2, Mob1, LATS1 and YAP1 mutants) develop various types of sarcoma. Despite that, Hippo member
Accepted 28 May 2015
genes are rarely point mutated in human sarcomas. Instead, WWTR1-CAMTA1 and YAP1-TFE3 fusion genes are
Available online 4 June 2015
found in almost all cases of epithelioid haemangioendothelioma. Also copy number gains of YAP1 and other
Keywords:
Hippo members occur at low frequencies but the most likely cause of perturbed Hippo signalling in sarcoma is
Sarcoma the cross-talk with commonly mutated cancer genes such as KRAS, PIK3CA, CTNNB1 or FBXW7. Current Hippo
Rhabdomyosarcoma pathway-targeting drugs include compounds that target the interaction between YAP and TEAD G protein-
Hippo pathway coupled receptors (GPCR) and the mevalonate pathway (e.g. statins). Given that many Hippo pathway-
YAP modulating drugs are already used in patients, this could lead to early clinical trials testing their efficacy in differ-
TAZ ent types of sarcoma.
Fusion genes Crown Copyright © 2015 Published by Elsevier B.V. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
2. Pathology of soft tissue sarcomas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
3. Hippo pathway & cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
4. Hippo mouse models that develop sarcomas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
5. Genetics of soft tissue sarcoma and the Hippo pathway. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
5.1. Hippo fusion genes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
5.2. Hippo copy number alterations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125
5.3. Epigenetic regulation of Hippo genes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125
5.4. Hippo signalling and the genomic landscape of sarcoma . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125
6. Hippo-targeted therapies in soft tissue sarcoma?. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
6.1. Drugs that target YAP-TEAD interaction or TEADs: verteporfin, cyclic peptides, VGLL4-mimicking peptides . . . . . . . . . . . . . . . . 126
6.2. G protein-coupled receptors (GPCRs) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
6.3. Statins and bisphosphonates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
7. Summary and perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
Transparency document . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
Funding. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

1. Introduction be conserved in mammals. In flies, inactivation of Hippo members re-

The Hippo pathway was discovered as a result of tumour suppressor
screens in the fruit fly (Drosophila melanogaster) and was later found to
⁎ Corresponding author.
E-mail address: h.wackerhage@abdn.ac.uk (H. Wackerhage).
sulted in an overgrowth that resembles the skin of a Hippopotamus,
thereby naming the pathway [1]. Hippo pathway members are rarely
point mutated in cancer [2,3] but the experimental mutation of Hippo
members usually causes overgrowth in fruit flies [4] and tumours in
mice, including sarcomas [2,3,5]. Here, we review the role of the Hippo
pathway specifically in soft tissue sarcomas excluding osteosarcomas

http://dx.doi.org/10.1016/j.bbcan.2015.05.006
0304-419X/Crown Copyright © 2015 Published by Elsevier B.V. All rights reserved.
122 A.D. Mohamed et al. / Biochimica et Biophysica Acta 1856 (2015) 121–129
and viral-mediated sarcomas such as Kaposi's sarcoma. After introducing
sarcomas and the Hippo pathway, we review animal models where
Hippo pathway-related transgenesis results in sarcomas. We then dis-
cuss findings and hypotheses implicating the Hippo pathway in sarcoma
signal transduction, development and pathology. Finally, we review the
current possibilities for Hippo pathway-targeting treatments in sarcoma
and highlight areas for future research.
2. Pathology of soft tissue sarcomas
Soft tissue sarcomas (from Greek sarx flesh) represent a biologically,
clinically, pathologically, and genetically [6] diverse array of malignant
tumours arising mostly from mesenchyme-derived tissues. Sarcomas
can occur in both adults and children. They are rare cancers and account
for about 1% of solid tumours in adults and for a significantly higher pro-
portion of cases in children (Cancer Research UK, 2015). In 2014, it was
estimated that 790 new cases of rhabdomyosarcoma and bone tumours,
including osteosarcomas and Ewing sarcomas, would be diagnosed in
children 0–14 years old, representing 7% of all childhood cancers
(American Cancer Society, 2014). The most common types of soft tissue
sarcomas include leiomyosarcoma, liposarcoma, fibrosarcoma, rhabdo-
myosarcoma and angiosarcoma (see Fig. 1 [7]).
The current classification of soft tissue sarcomas is based on a com-
bination of tumour morphology, immunophenotype and molecular pa-
thology. Genetically, sarcomas can show either aberrant, chimeric
transcription regulators as a consequence of fusion genes such as
PAX3/7-FOXO1 [6,8], somatic point mutations of well-known cancer
genes such as oncogenic RAS isoforms, PIK3CA or TP53, or DNA copy
number gains or losses [6,9–12]. As we will show later, there is no evi-
dence for recurrent Hippo gene point mutations in sarcomas, while
there is abundant evidence for mutations of typical cancer genes [13,
14] that can cross-talk to the main members of the Hippo pathway. Ad-
ditionally, fusion genes involving WWTR1 or to a lesser extent YAP1 are
found in nearly all cases of epithelioid haemangioendothelioma [15,16],
whereas YAP1 and VGLL3 copy number gains have been reported for
some types of sarcoma, especially rhabdomyosarcoma [17–19]. It is un-
clear to this date whether VGLL3, which is associated with tumour sup-
pression in ovarian cancer [20], is a bona fide Hippo pathway member.
The increasing use of immunohistochemistry and the application of
sophisticated molecular techniques combined with a better under-
standing of soft tissue sarcoma biology have led to a continued refine-
ment of the classification of soft tissue sarcomas. Some previously
recognised types of soft tissue sarcoma, most notably malignant fibrous
histiocytoma, are now being reclassified. Most tumours previously clas-
sified as malignant fibrous histiocytomas but showing no specific
immunophenotype would now be regarded as undifferentiated pleo-
morphic sarcomas. The majority of soft tissue sarcomas arise in the
limbs of older people although rhabdomyosarcoma, especially the
Dedifferentiated
Pleomorphic Myxoid
Well differentiated
Liposarcoma Biphasic
Gastrointestinal
(adipocytes) Synovial sarcoma
stromal tumour (GIST) Spindle cell
Epithelioid haemangio-
endothelioma
Leiomyosarcoma Soft tissue sarcoma Vascular
(smooth muscle) Angiosarcoma
Kaposi’s sarcoma
Rhabdomyosarcoma
(myoblasts)
Fibrosarcoma
(fibroblasts)
Low-grade
Embryonal fibromyxoid sarcoma
Spindle cell
Alveolar
Pleomorphic Myxofibrosaroma
Fig. 1. Classification of soft tissue sarcomas on the basis of their differentiation.
embryonal subtype, tends to occur predominantly in young children
[21]. Most soft tissue sarcomas arise sporadically but muscle injury,
most likely by increasing the number of activated satellite cells, en-
hances the penetrance and shortens the latency of rhabdomyosarcoma
phenotypes in mice [18,22]. In addition, known risk factors for
the development of specific types of sarcoma in humans include expo-
sure to radiation (e.g. post-radiation angiosarcoma) or exposure to en-
vironmental/occupational carcinogens (e.g. vinyl chloride-associated
angiosarcoma). Sarcomas, in contrast to carcinomas, have a predilection
for showing a vascular pattern of spread with the lungs generally being
one of the predominant sites of metastasis.
A combination of surgery, cytotoxic chemotherapy and radiotherapy
are the mainstays of current treatment with some types of sarcoma
being treated with neoadjuvant chemo-radiotherapy prior to definitive
surgery [23–25]. Immunotherapy and targeted, novel biologic therapies
are also now being evaluated in the treatment of specific types of sarco-
ma [26–29] and as we show below, targeting the Hippo pathway in sar-
coma should already be possible with existing drugs.
The prognosis and outcome of soft tissue sarcomas depend on a
range of factors including the particular subtype of tumour, the anatom-
ical location, size and grade as well as tumour stage at time of diagnosis
[7,30,31]. The overall 5-year survival rate for soft tissue sarcomas has
gradually been improving, especially in children, but the outcome varies
with the specific type of soft tissue sarcoma and depends on the prog-
nostic factors that have been outlined above. Overall the 5-year survival
rate for adults with soft tissue sarcomas is approximately 60% while it is
higher in children, reaching approximately 70% (Cancer Research UK,
2015).
3. Hippo pathway & cancer
The fruit fly (D. melanogaster) has been used over many decades as a
model organism to identify genes whose knockout results in cancerous
growth [32]. This research has led to the discovery of a set of genes that
encode two interacting kinases and auxiliary proteins, now defined as
the core Hippo pathway (see Fig. 2). The main function of the Hippo
pathway is to inhibit proliferation and to promote apoptosis, thereby
limiting organ growth [4]. In the conserved mammalian Hippo pathway,
the STE20-like protein kinases 1 and 2 (MST1 and MST2, gene symbols:
STK4 and STK3) regulate the large tumour suppressor kinases 1 and 2
(protein name and gene symbol: LATS1 and LATS2), through phosphor-
ylation [33]. Active LATS1 and LATS2 then interact through their PPxY
motifs with the WW domains of the transcriptional co-factors YAP
(gene symbol YAP1) or TAZ (gene symbol WWTR1; note that the gene
TAZ encodes a protein termed Tafazzin which is not part of the path-
way) [34]. This physical contact allows LATS1 and LATS2 to inhibit
YAP [35] and TAZ [36] through the phosphorylation of multiple
HXRXXS amino acid motifs. The phosphorylation of these motifs pro-
motes the inactivation of YAP and TAZ through translocation from the
nucleus into the cytosol and degradation. Additionally, YAP can be phos-
phorylated at Tyr357 by the tyrosine kinase YES1, which has resulted in
its name Yes-associated protein (YAP) [37]. Nuclear and active YAP,
which was first discovered by Marius Sudol [38,39], and its paralogue
TAZ is believed to exert their tumourigenic functions mainly via the
TEAD transcription factors [40]. Specifically, YAP and presumably TAZ
de-repress and activate the TEAD transcription factors that otherwise
recruit transcriptional repressors [41]. Additionally, YAP and TAZ are ca-
pable of co-regulating other transcription factors including those be-
longing to the Smad family [42] and Tbx5 in some contexts [37,43]. In
addition to the Hippo kinases, extensive cross-talk mechanisms also
regulate the activity of YAP and TAZ, notably mechanotransduction
[44], WNT signalling [45,46], and G protein-coupled receptors [47].
The early studies in fruit flies and subsequent studies in mammals
demonstrate that the upstream proliferation-inhibiting Hippo proteins
and the proliferation-promoting Hippo transcriptional regulators
act as potent tumour suppressors and oncogenes, respectively. For
 A.D. Mohamed et al. / Biochimica et Biophysica Acta 1856 (2015) 121–129 123

Fig. 2. Schematic drawing depicting proteins associated with the Hippo pathway in sarcoma. A The Hippo kinases MST1/2 and LATS1/2 together with the auxiliary proteins SAV1 and Mob1
inhibit the transcriptional co-factors YAP and TAZ by phosphorylation of HXRXXS motifs such as Ser127 on YAP. B YAP and TAZ dephosphorylation by phosphatases results in their nuclear
translocation, allowing the de-repression or activation of TEAD1-4 transcription factors. In some contexts, VGLL4 acts as a co-repressor of TEAD1-4 [41] and other VGLL isoforms are likely
to be capable of interaction with TEADs [105]. C–G Links between the Hippo pathway and proteins whose genes are recurrently mutated in sarcoma (shown in red) C In epithelioid
haemangioendothelioma, mostly the WWTR1-CAMTA1 [16] but also YAP1-TFE3 [15] fusion genes (encoding TAZ-CAMTA1 and YAP-TFE3 fusion proteins) presumably regulate the TEAD
transcription factors. D The Wnt-regulator β-CATENIN (CTNNB1) can also interact with YAP and TAZ. It has been proposed that β-CATENIN, YAP, the tyrosine kinase YES1 and the tran-
scription factor TBX5 form complexes of functional importance in cancer. In this context, the tyrosine kinase YES1 can activate YAP by Tyr357 phosphorylation [37]. E KRAS and PI3K
can enhance the co-activator function of YAP, or their activity can be substituted by YAP to overcome oncogene addiction, identifying YAP as a modulator of drug resistance. F In alveolar
rhabdomyosarcoma the PAX3-FOXO1 fusion proteins/genes drive the expression of RASSF4 which inhibits MST1 [84]. Not shown: Others suggest that YAP and TAZ are included in the Wnt
destruction complex [46]. G The SCF (βTRCP) ubiquitin ligase regulates YAP [35] and especially TAZ [94] degradation via phosphodegron-mediated mechanisms. Recently another ubiq-
uitin ligase named FBXW7, which is frequently mutated in rhabdomyosarcoma [73], was identified as part of a complex that mediates ubiquitin-dependent destruction of YAP and TAZ and
its loss was associated with increased YAP abundance [95]. Drugs and drug targets that modulate the activity of the Hippo pathway are shown as red text. Verteporfin [97] and cyclic pep-
tides [106] inhibit the binding of YAP to TEAD transcription factors. VGLL4-mimicking peptides repress TEAD transcription factors [79]. Specific G protein-coupled receptors (GPCRs) signal
to YAP [47]. Given that GPCR-targeting drugs are the largest class of currently used drugs some might be applicable for Hippo activity modulation in sarcomas. The activity of YAP and TAZ is
regulated by the mevalonate pathway [102,103], which can itself be targeted for example by statins. For more Hippo pathway related drugs see [107].

example, YAP hyperactivity as a consequence of the sole expression of
the constitutively active YAP1 S127A mutant in mice alone is sufficient
to initiate hyperplasia and over time cause cancer in the liver [48,49]
and skin [50]. Also, along with injury in the skeletal muscle, YAP1
S127A expression in satellite cells alone is sufficient to causes rhabdo-
myosarcoma [18]. These studies identify YAP as an unusually potent on-
cogene. Thus it would be intuitive that YAP-activating mutations such as
YAP1 S127A occur by chance and be a driver of tumourigenesis. Howev-
er, this is not the case. So far, even if the LATS kinases appear to be mu-
tated in some rare cases [51], recurrent mutations of Hippo pathway
genes have not been reported [2,3,5,13]. It is even more surprising
that not a single YAP1 S127A mutation was found in 21,441 tumour
DNA samples from 91 cancer studies that are compiled in the cBIOPortal
database [52,53].
From this conundrum two questions arise: First, why are Hippo
genes, unlike other well established cancer-associated genes such as
TP53, PIK3CA or KRAS [13], rarely point mutated in cancer? Second, if
point mutations are not major the source of tumourigenic Hippo signal-
ling, what else is causing the high YAP activity frequently seen in
cancers? A potential answer to the first question is that the somatic
(i.e. cancer cell) mutation frequency differs at least 100-fold in-
between different types of cancer and also varies across the genome
from ≈ 0.1 to ≈ 10 mutations per 100,000,000 bases depending on
DNA replication timing and transcriptional activity [54,55]. There is ev-
idence that copy number gains and other rearrangements are especially
found in early replicating chromatin [55]. Thus, Hippo genes may lie in
chromatin with a low mutation rate perhaps due to their functional im-
portance. Regarding the second question, alternative mechanisms of
YAP and TAZ activation in cancer include Hippo fusion genes, copy num-
ber alterations of Hippo genes, hypermethylation of Hippo kinase gene
promoters and cross-talk from significantly mutated non-Hippo cancer
genes. Most prominently, Hippo fusion genes have been identified for
almost all cases of epithelioid haemangioendothelioma [15,16], a vascu-
lar sarcoma that will be discussed below. Copy number gains have been
reported for YAP1 and VGLL3 [17,18,56] and frequently mutated cancer
genes such as CTNNB1 (protein: β-catenin [37] or KRAS [57–59] have
been shown to affect YAP activity and require YAP for some aspects of
tumourigenesis.
4. Hippo mouse models that develop sarcomas
Important direct evidence for a role of the Hippo pathway in sarco-
ma stems from transgenic Hippo mouse models that develop sarcoma.
Such models are summarised in Table 1 and discussed in greater detail
below.
As discussed earlier, the Hippo pathway kinase Lats1 inhibits YAP
and TAZ by phosphorylation and restricts their nuclear localization
and transcriptional activity with the TEAD factors. Most Lats−/− mice
die in utero, and only ~8% survive. The surviving mice display a strong
growth retardation phenotype, severe fertility defects and pituitary
124 A.D. Mohamed et al. / Biochimica et Biophysica Acta 1856 (2015) 121–129
Table 1
Hippo transgenic animal models that lead to the development of sarcomas.
Genotype Sarcoma type
Lats1−/− Skin fibrosarcoma
Lats1−/− and mutagen (DMBA+ UVB exposure) Skin fibrosarcoma
Nf2/Merlin +/− Osteosarcoma***
Fibrosarcoma*
Chondrosarcoma
Uterine sarcoma
Nf2/Merlin +/− p53 +/− (cis)** Osteosarcoma***
Fibrosarcoma*
Nf2/Merlin +/− p53 +/− (trans)** Osteosarcoma***
Fibrosarcoma*
Mob1aΔ/+1btr/tr or Mob1aΔ/Δ1btr/+ Extraskeletal osteosarcoma, fibrosarcoma†
Pax7CreER TetO-YAP1 S127A and cardiotoxin injury Embryonal rhabdomyosarcoma
*Of note: In this study, the tumours were reported to also contain areas presenting features of rhabdomyosarcoma within the same tumours, but were classified predominantly as
fibrosarcomas.
**p53 mutations were introduced on the same allele as the NF2/Merlin mutation (in cis), or on the other allele (in trans).
***In nearly all sarcomas, loss of the other wild-type NF2/Merlin allele was observed in the tumours (Loss of Heterozygosity, LOH);
†
or myofibrosarcoma.
hyperplasia leading to hormonal impairment. Indeed, Lats1−/−
females have reduced levels of luteinizing hormone, prolactin and
growth hormone. All Lats1−/− females have underdeveloped mammary
glands and develop ovarian stromal cell tumours by 3 months of age.
Large skin fibrosarcomas develop in 14% of all Lats1−/− females be-
tween 4-10 months of age. The incidence of skin fibrosarcomas in-
creases to 83% when LATS1−/− mice are subjected to a mutagen
treatment consisting of a single application of 9,10-dimethyl-1,2-benz-
anthracene (DMBA) followed by repeated exposure to UVB, whereas all
Lats1+/− animals remained tumour-free [60].
Neurofibromatosis type II is a dominantly inherited disorder caused
by mutations in the neurofibromin 2 (gene symbol Nf2) gene, encoding
a protein called Merlin, also known as schwannomin. In both drosophila
and mammals, Nf2/Merlin acts upstream of the Hippo kinases to control
the transcriptional activity of Yap [63,64]. Mutations in the Nf2 gene
identified in Neurofibromatosis type II patients lead to the production
of shorter protein products that have lost the tumour suppressor
function associated with full-length Merlin. Consequently, neurofibro-
matosis type II patients are predisposed to developing tumours affecting
the nervous system, primarily schwannomas, meningiomas and
ependymomas [61]. However, mice expressing a mutated Nf2 allele,
originally intended to mimic human Neurofibromatosis type II in
mice, do not develop schwannomas, meningiomas or ependymomas.
Instead, they principally develop osteosarcomas (63%), fibrosarcomas
(9%) and hepatocellular carcinomas (9%) occurring between 10 and
30 months of age [61]. Notably, human neurofibromatosis type II pa-
tients do not develop osteosarcomas, fibrosarcomas or hepatocellular
carcinomas at higher incidence than the normal population [61]. Also,
chondrosarcomas and uterine sarcomas arose at low frequency in
Nf2+/− mice, and in some cases loss of the wild-type Nf2 allele was de-
tected [61]. However, nearly all of the osteosarcomas and fibrosarcomas
displayed a loss of the wild-type Nf2 allele (LOH), supporting a role for
the complete loss of Merlin function in the aetiology of those tumour
types. These tumours also displayed an enhanced metastatic potential
[61]. An additional hemizygous inactivation of p53 produced a similar
tumour spectrum but enhanced the incidence of sarcomas (osteosarco-
mas, 77%; fibrosarcomas, 32%) and significantly reduced tumour-free
survival [61].
Mob1 proteins are core components of the Hippo tumour suppressor
pathway that are required for activation of the LATS kinases by the MST
kinases [65]. Mice bearing a null mutation of the Mps one binder (Mob)
kinase activator 1A (Mob1aΔ/Δ) or a trapped mutation of the Mob1b
gene (Mob1btr/tr) show no abnormalities in morphology, body weight,
histology, or life span [62]. While the complete loss of both genes
(Mob1a and Mob1b; Mob1aΔ/Δ1btr/tr) is embryonic lethal, double-
mutant mice retaining one allele of either Mob1a (Mob1aΔ/+1btr/tr) or
Incidence Latency Reference
14% 4–10 months [60]
83% 10–14 weeks after birth and treatment [60]
63% 10–30 months [61]
9% 10–30 months
Low frequency Not determined
Low frequency Not determined
77% b5 months [61]
32% b5 months
74% 4–21 months [61]
8% 4–21 months
24% 25–70 weeks [62]
22% 25–70 weeks
100% 10–11 weeks after cardiotoxin injury [18]
Mob1b (Mob1aΔ/Δ1btr/+) survive. Of these heterozygotes, 52% develop
dental malocclusion, 25% have a disorganization of the inner ear hair
bundles in the cochlear organ of Corti resulting in a disequilibrium
and 31% display increased trabeculae in femurs [62]. In addition, 100%
of the Mob1aΔ/+1btr/tr and Mob1aΔ/Δ1btr/+ heterozygotes mice sponta-
neously develop various types of tumours, while only 4% of the control
mice (heterozygotes for both genes; Mob1aΔ/+1btr/+) developed tu-
mours [62]. All single heterozygote mice developed skin cancer
(100%), 92% developed benign bony overgrowths called exostoses,
24% developed extraskeletal osteosarcomas and 22% developed subcu-
taneous fibrosarcomas or myofibrosarcomas. These mice also devel-
oped other tumour types at lower incidence, such as breast (16%),
lung (5%) and salivary gland (5%) tumours. Interestingly, while 50% of
the Mob1aΔ/Δ1btr/+ mice developed liver tumours, not a single liver tu-
mour was found in the Mob1aΔ/+1btr/tr mice [62].
YAP, along with its paralogue TAZ, is the main transducer of the
Hippo pathway activity via its de-repression and activation of TEAD
transcription factors. Specifically in activated but not quiescent satellite
cells, the overexpression of YAP1 S127A alone is sufficient to cause em-
bryonal rhabdomyosarcomas in 100% of the mice [18], in line with the
pro-proliferative and anti-differentiation roles of YAP in myoblasts
and primary satellite cells in culture [66,67]. The mouse tumours
match human ERMS well as judged by their pathology and gene expres-
sion profile. Conversely, YAP1 knockdown in the human RD ERMS cell
line reduces their proliferation, soft agar growth and xenotransplant
burden as well as promotes myogenic differentiation [18]. The fact
that YAP1 S127A drives tumourigenesis only in activated but not quies-
cent satellite cells matches clinical observations. First, ERMS occurs es-
pecially in infants and young children where activated satellite cells
support longitudinal muscle growth [21]. Additionally, mdx mice,
where many satellite cells are activated to regenerate damaged muscle
[68] spontaneously develop rhabdomyosarcomas [69]. Dystrophin and
dysferlin double-mutants are also emerging as potential models for
rhabdomyosarcoma [70].
5. Genetics of soft tissue sarcoma and the Hippo pathway
In this section we will review Hippo-related genetic changes in sar-
comas. Specifically, we discuss Hippo fusion genes, Hippo copy number
changes, epigenetic alterations of Hippo genes and mutations that may
affect Hippo members through cross-talk in sarcomas.
5.1. Hippo fusion genes
Fusion products involving Hippo genes have been associated
especially with epithelioid haemangioma, a vascular sarcoma. In this
 A.D. Mohamed et al. / Biochimica et Biophysica Acta 1856 (2015) 121–129
tumour, the WWTR1-CAMTA (WWTR1 is the gene symbol for TAZ) fu-
sion product was found in 17 cases independent of location [16]. A
follow-up analysis on the 10% of epithelioid haemangiomas that were
negative for the WWTR1-CAMTA identified those as a pathologically dis-
tinct subset. In eight of these cases a YAP1-TFE3 fusion product was
found [15]. These findings were confirmed by a study on 35 epithelioid
haemangioendotheliomas in which WWTR1-CAMTA fusions were found
in 33 cases and YAP1-TFE3 fusion genes in the remaining 2 cases [71]. In
this type of tumour, fusion products involving Hippo genes appear to be
as specific as the PAX3/7-FOXO1 fusions are for alveolar rhabdomyosar-
comas [72,73]. Apart from epithelioid haemangioma, a single case of
spindle cell rhabdomyosarcoma was associated with a TEAD1-NCOA1
fusion protein [74]. How these Hippo fusion genes contribute to
tumourigenesis and why epithelioid haemangioendotheliomas appear
to depend almost in every case on WWTR1-CAMTA or YAP1-TFE3 fusion
genes is unknown. Recently, the WWTR1-CAMTA fusion was shown to
inhibit the tumour suppressive function of the CAMTA transcriptional
regulator and to confer a TAZ-like transcriptional programme in cells
expressing the fusion, thereby favouring oncogenic transformation
and resistance to anoikis [75].
5.2. Hippo copy number alterations
Generally, specific types of cancer are either dominated by high
levels of somatic point mutations (termed M type) or by copy number
gains with TP53 mutations (termed C type), or by intermediate but
not high levels of both [76]. This may reflect different causes of muta-
genesis and also be related to chromatin features, as point mutations
and copy number losses are associated with late replicating chromatin
whereas copy number gains and rearrangements are associated with
early replicating chromatin [54,55]. Generally, Hippo genes appear far
more affected by copy number gains and rearrangements than by
point mutations as evidenced from Hippo genes analyses using the
cBioPortal. This supports the idea that Hippo genes are possibly located
in early replicating chromatin, consistent with the elevated copy num-
ber alterations found in C type cancers and the existence of YAP1 and
WWTR1 fusion genes. Indeed, sporadic Hippo copy number gains are
found in different types of sarcoma (see Fig. S1 for a re-analysis of the
data of Barretina et al. 2010 using the cBIOPortal platform [77]). Copy
number gains of YAP1 (11q) were reported in 22 cases and copy number
gains of VGLL3 (3p) were found in 19 cases of soft tissue sarcoma sub-
types [17]. YAP1 copy number gains were additionally observed in em-
bryonal but not alveolar rhabdomyosarcoma [18] and high level VGLL3
amplifications were found in 10 out of 12 cases of myxoinflammatory,
fibroblastic sarcoma and hemosiderotic, fibrolipomatous tumours [78].
Additionally the re-analysis of cBioPortal-deposited sequencing data
for different types of sarcoma [77] reveals YAP1, WWTR1, VGLL1-4 or
TEAD1-4 copy number gains but no losses in 17% of the cases (YAP1
3%, WWTR1 2%, VGLL4 2%, VGLL3 5%; Fig. S1). However, it is currently un-
known, although seemingly unlikely, whether all of these copy number
gains in Hippo genes are bona fide cancer drivers and whether their ef-
fect is a major one. Indeed, copy number gains of YAP1, VGLL3, VGLL4 or
TEAD1-4 appear to co-exist even though VGLL4 has been associated
with a disruption of TEAD/YAP transcriptional activity and tumour sup-
pression [41,79]. Second, high YAP activity appears to result in the acti-
vation of a negative feedback loop [67]. In line with this, a higher
concentration of YAP (or elevated YAP nuclear localization and tran-
scriptional activity) is generally associated with an increase in the levels
of inhibitory YAP Ser127 phosphorylation because of the increased ex-
pression of several members of the Hippo kinase cascade that inhibits
YAP through phosphorylation [67]. This is functionally supported by
the fact that the expression of a wild-type YAP1 does not prevent
myogenic differentiation whereas the expression of constitutively ac-
tive YAP1 S127A does prevent differentiation [66]. Collectively, this sug-
gests that recurrent copy number alterations, especially gains, in the
TEAD co-regulator genes YAP1, WWTR1, VGLL3 and VGLL4 in a subset
125
of sarcomas can only partially explain why the balance is tipped to-
wards tumourigenesis.
5.3. Epigenetic regulation of Hippo genes
Epigenetically mediated loss of the expression of the Hippo ki-
nases or other Hippo pathway components has been associated
with certain cancers [80,81]. In primary soft tissue sarcomas, the
CpG island in of MST1 and MST2 kinase promoters were found to be
significantly hypermethylated. MST1 was methylated in liposarcoma
(25%), leiomyosarcoma (50%), malignant fibrous histiocytoma and
myxofibrosarcoma (17%) as well as in rhabdomyosarcoma (80%). In
contrast MST2 was found methylated principally in liposarcoma (15%),
malignant fibrous histiocytoma and myxofibrosarcoma (31%) as well
as synovial sarcoma (50%). MST1 methylation was associated with a de-
creased expression of the MST1 mRNA, and surprisingly, a significantly
increased risk for tumour-related death was found for patients with
an unmethylated MST1 promoter. The tumour suppressor kinase
LATS1 was also methylated in 7% of all sarcomas tested and the LATS2
and WW45/Salvador genes were not found methylated in sarcomas.
The promoter of the tumour suppressor gene RASSF1A has been shown
to be methylated in liposarcoma (18%), leiomyosarcoma (39%), malig-
nant fibrous histiocytoma and myxofibrosarcoma (6%) and in neurogenic
sarcomas (50%), which is associated with an unfavourable prognosis for
cancer patients [82]. These results, obtained with a small sample size,
are a proof-of-principle for the epigenetic regulation of Hippo genes. It
remains to be determined if the methylation status of the MST kinases
is linked with YAP activity in sarcomas. Indeed, the MST kinases have
been shown to be dispensable for Hippo pathway activity in some cell
types, notably in mouse embryonic fibroblasts and skin [83].
5.4. Hippo signalling and the genomic landscape of sarcoma
So far two studies have reported the sequencing of multiple types of
sarcoma [77] and of rhabdomyosarcoma, a sarcoma where skeletal
muscle-related genes are expressed [73]. Additionally, preliminary
data from a TCGA sarcoma study are accessible through cBIO but these
data are under publication embargo until 2016. In the earliest large-
scale sequencing study 207 high grade, seven types of soft-tissue sarco-
mas were analysed for the expression of 722 protein-coding and
microRNA genes [77]. A limitation of this study was the subjective selec-
tion of the genes sequenced and the relatively small sample number for
such an analysis. However, the mutated genes CTNNB1, PIK3CA, NF1 and
PIK3CA identified in this study were also identified later in a larger unbi-
ased study involving next generation sequencing of rhabdomyosar-
comas [73].
In rhabdomyosarcoma, the most distinct subgroup is alveolar rhab-
domyosarcoma (ARMS). This subtype is in most cases marked by
PAX3-FOXO1 or PAX7-FOXO1 fusion genes with rare alternative PAX fu-
sions [73]. A recent study has linked PAX3-FOXO1 to the increased ex-
pression of RASSF4 which inhibits the Hippo kinase MST1 [84]. Such
an inhibition of a Hippo kinase should result in an increased transcrip-
tional output downstream of the Hippo signalling cascade, but the
exact mechanism could not be identified. Nevertheless, this suggest
cross-talk between the ARMS PAX3/7-FOXO1 fusions and Hippo signal-
ling. Conversely, the rhabdomyosarcomas that are negative for PAX3/
7-FOXO1 fusions, which represent mostly embryonal rhabdomyosar-
comas and 20% of the alveolar RMS cases [85], carry point mutations
of cancer genes that are frequently mutated pan-cancer [13] including
KRAS, HRAS, NRAS, NF1, PIK3CA, CTNNB1 and FBXW7 [73]. Many of
these mutated cancer genes are likely to cross-talk to Hippo members
(described in more detail below). Additionally, the genomes of ERMS
are complex and are aneuploid (i.e. changed chromosome numbers)
and display segmental aberrations whereas the genomes of ARMS
only show few of such changes [73,86]. While the genes affected in
ERMS vary greatly, 171 out of 196 ERMS tumours were positive for
126 A.D. Mohamed et al. / Biochimica et Biophysica Acta 1856 (2015) 121–129
YAP by immunohistochemistry and in 143 out of these 171 cases YAP
was nuclear [18]. Additionally, the expression of constitutively active
YAP1 S127A in mice resulted in the formation of ERMS-like tumours
with short latency in every case [18]. This suggests that several of the
mutated ERMS genes [73], together with the YAP1 copy number gains
[17,18], converge to towards increasing YAP activity in ERMS, which is
likely driving the disease.
There is evidence that several of the somatically point mutated
ERMS and sarcoma genes can cross-talk to YAP. Several recent reports
have linked KRAS to YAP in cancer. To identify genes that can substitute
for oncogenic RAS, 15,294 genes were expressed in KRAS-dependent
colon cancer cells. This non-biased analysis revealed that YAP could res-
cue the viability of KRAS-addicted cells and that YAP was required for
KRAS-induced cell transformation [57]. In another study, the authors ob-
served in a mouse model of pancreatic ductal adenocarcinoma that
some KRAS G12D-driven tumours relapsed after KRAS inhibition. The re-
sultant tumours exhibited an amplification and overexpression of YAP1
[58]. YAP1 was also identified in a non-biased screen using 27,500
shRNAs to knockdown 5046 signalling genes as the top hit for a gene re-
sponsible for resistance to RAF or MEK inhibitor therapy [87]. Also, in
pancreatic ductal adenocarcinoma, it was found that YAP1 expression
was increased and that the deletion of YAP1 prevented the progression
of early lesions [59]. Taken together, it seems likely that similar mecha-
nisms operate in ERMS where oncogenic RAS mutations (NRAS, KRAS,
HRAS or mutations of the RAS inhibitor NF1) are relatively common
[73]. In line with this, the knockdown of YAP in the RD ERMS cell line,
which carries a NRAS Q61H mutation (accompanied by MYC amplifica-
tion and TP53 mutation with a karyotype of 51-hyperdiploidy [88]), re-
duces proliferation, soft agar growth, and xenotransplant tumour
burden while promoting the differentiation of these cells [18].
Somatic mutations of PIK3CA, which encodes phosphoinositide 3-
kinase (PI3K), have been reported in 14% of all cases in a large-scale
pan-cancer study [13] showing that PI3K is a frequently mutated gene
pan-cancer. It has been demonstrated that epidermal growth factor
(EGF) inhibits the Hippo pathway through PI3K, resulting in YAP nucle-
ar localisation transcriptional activity [89]. In this study, it has been sug-
gested that this effect is independent of PKB/AKT, which is a key kinase
downstream of PI3K. However, several studies have demonstrated
cross-talk between Hippo signalling and AKT1, which can phosphory-
late MST1 at Thr120 and Thr387 leading to the inhibition of Mst1
activity [90–92]. Thus, AKT1-MST1 cross-talk could be an alternative
mechanism by which PIK3CA mutations affect the activity of YAP or TAZ.
Mutations of the Wnt gene CTNNB1 (β-catenin) occur in some ERMS
cases [73]. We have already mentioned the close integration of Wnt and
Hippo signalling. Specifically, it was demonstrated that YAP was crucial
for the proliferation and soft agar growth of cancer cell lines with high
β-catenin activity [37]. It was shown that β-catenin can form complexes
with YES1, YAP and TBX5 [37]. Intriguingly, a loss-of-function screen
implicated YES1 in the survival of rhabdomyosarcoma cells [93], per-
haps suggesting that similar mechanisms operate in sarcomas.
The SCF (β-TrCP) ubiquitin ligase regulates YAP [35] and TAZ [94]
degradation via well-defined phosphodegron-mediated mechanisms.
FBXW7 is another E3 ubiquitin ligase that was suggested to regulate
the degradation of YAP [95]. Indeed, an inverse correlation was reported
between FBXW7 and YAP protein abundance in hepatocellular carcino-
ma. Furthermore, YAP overexpression could counteract the apoptosis
and growth arrest caused by FBXW7 [95]. This effect of FBXW7 on YAP
levels is consistent with the notion that a loss of FBXW7 in ERMS could
favour a higher level of YAP activity. Collectively, this also implies that
Hippo signalling plays an important but different role in both ARMS
[84] and ERMS downstream of the mutated genes [18].
6. Hippo-targeted therapies in soft tissue sarcoma?
Before 2012, the Hippo pathway was considered un-druggable, but
since 2012 several compounds and treatment strategies have emerged.
Generally, these drugs can be sub-divided into either drugs that target
the binding of YAP to TEAD transcription factors or into drugs that mod-
ulate other members of the wider Hippo signal transduction network,
such as G protein-coupled receptors or the mevalonate pathway. Sever-
al potential Hippo drugs such as verteporfin or statins are already wide-
ly used in patients to treat other conditions and this can pave the way
for early clinical trials involving such compounds. Key obstacles for sys-
temic and long-term Hippo-targeting treatments for sarcoma, and for
cancer in general, are that the Hippo pathway is ubiquitously expressed
(i.e. the drugs may perturb signalling in other tissues and cause side ef-
fects), although, in many tissues, the loss-of-function of YAP is incon-
spicuous under homeostatic conditions until tissue regeneration is
required. Here we discuss the different Hippo treatment options that
might become useful for the treatment of sarcomas that result from dys-
regulated Hippo signalling.
6.1. Drugs that target YAP-TEAD interaction or TEADs: verteporfin, cyclic
peptides, VGLL4-mimicking peptides
Verteporfin, a member of the porphyrin family, contains aromatic
heterocyclic molecules formed of four modified pyrrole units intercon-
nected at their carbon atoms by methane bridges. Verteporfin is already
a clinically approved photosensitizer for vascular macular degeneration
[96]. Verteporfin binds to YAP and changes its conformation thereby
inhibiting its interaction with TEAD2 and, presumably, with other
TEAD isoforms. Furthermore, verteporfin decreased liver overgrowth
and blocked liver tumourigenesis induced by YAP overexpression or en-
dogenous YAP activation [97]. Moreover, uveal melanoma is a cancer
resulting from gain-of-function mutations in the GNAQ and GNA11 on-
cogenes encoding the heterotrimeric Gαq family that stimulate YAP ac-
tivity. Verteporfin has also been shown to reduce the tumour formation
potential of GNAQ/GNA11 mutated uveal melanoma cells in xenograft
assays [98,99].
6.2. G protein-coupled receptors (GPCRs)
G protein-coupled receptors (GPCRs) are the largest family of cell
surface receptors. Deep sequencing studies have shown an association
between aberrant expression and activity of GPCRs and tumourigenesis
in nearly 20% of human cancers carrying mutations in GPCRs [100]. Re-
cent studies have revealed that the Hippo pathway can be broadly reg-
ulated by different GPCRs. Serum-borne lysophosphatidic acid (LPA)
and sphingosine 1-phosphate (S1P) inhibit the Hippo kinases LATS1/2
by acting through G12/G13 coupled receptors and Rho leading to induc-
tion of YAP/TAZ transcription activity [47]. Therefore, the drugs
targeting the GPCR isoforms affecting Hippo signalling represent a po-
tential therapeutic approach to treat sarcomas driven by high activity
of YAP or TAZ.
6.3. Statins and bisphosphonates
Aberrant activity of the mevalonate pathway can promote tumour
progression in vivo and anchorage independent growth of cells
in vitro. Also, high mRNA levels of hydroxymethylglutaryl coenzyme A
reductase (HMGCR) and mevalonate pathway genes are associated
with poor prognosis and reduced survival in breast cancer patients
[101]. Two recent studies have shown that YAP/TAZ activity can be reg-
ulated by the mevalonate pathway [102,103]. Mechanistically, the
geranylgeranyl phosphate produced by the mevalonate cascade acti-
vates Rho GTPases, which in turn increases YAP/TAZ activity by
inhibiting their phosphorylation and enhancing their nuclear accumula-
tion. Interestingly, statins and bisphosphonates that inhibit distinct en-
zymes of the mevalonate pathway have a marked effect on YAP/TAZ
transcriptional activity. Taken together these findings suggest that
YAP/TAZ are presumably mediating the oncogenic responses associated
with the dysregulation of the mevalonate pathway. Moreover, statins
 A.D. Mohamed et al. / Biochimica et Biophysica Acta 1856 (2015) 121–129
and bisphosphonates are potential treatments to target the malignant
effects of YAP and TAZ in cancer cells [102,103]. In accordance with
this, a number of recent studies have reported a possible preventive or
therapeutic role of statins in cancer, and further studies are ongoing to
validate and characterize the anticancer effects of statins in various con-
texts [104].
7. Summary and perspectives
In summary, the Hippo pathway comprises potent tumour suppres-
sors and oncogenes that are rarely somatically point mutated in sarco-
mas or in other types of human cancer. The involvement of Hippo
members, especially YAP and TAZ, in human epithelioid haemangioma
occurs in the form of WWTR1-CAMTA and YAP1-TFE3 fusion genes,
whereas copy number alterations of YAP1 and other Hippo genes occur
generally at a low frequency in other types of sarcomas. However, it is
unclear whether these copy number alterations are important or even
essential for sarcomagenesis in the presence of functional Hippo kinases.
Thus, the majority of the changes in the activity of Hippo genes, and spe-
cifically YAP1, in sarcoma probably result from the cross-talk with sever-
al genes that are frequently mutated in sarcoma and across cancers. For
instance, the PAX3-FOXO1 fusion gene driving alveolar RMS can inter-
play with the Hippo pathway via the upregulation of RASSF4, a negative
regulator of the MST1 kinase, and perhaps also by modulating YAP1 ac-
tivity [84]. In other sarcoma subtypes like embryonal RMS, somatic gain-
of-function mutations in KRAS, PIK3CA or CTNNB1 and loss-of-function
mutations in FBXW7 could affect Hippo pathway activity. Perturbed
Hippo signalling can clearly drive sarcomagenesis as evidenced from
mouse models expressing YAP1 S127A (gain-of-function) or from mice
harbouring LATS1, Nf2 or Mob1a/b loss-of-function mutations. Also, the
near 100% penetrance of WWTR1-CAMTA or YAP1-TFE3 fusion genes in
epithelioid haemangioendotheliomas suggests that these Hippo fusion
genes are driving sarcomagenesis.
Given the apparent importance of Hippo signal transduction path-
way members in sarcoma it is welcome news that many different vali-
dated or putative Hippo treatments have emerged since 2012. They
can be subdivided into drugs that directly target the Hippo transcrip-
tional regulators and drugs that target other members of the wider
Hippo signal transduction network. Some of these drugs are already
widely used for the treatment of other diseases and this could facilitate
the progression towards preclinical studies aimed at inhibiting the ac-
tivity of YAP or TAZ in sarcomas.
Several lines of evidence now support the notion that the Hippo
pathway might be a major player in sarcomagenesis, but they also
raise a number of questions yet to be answered:
– Can the expression, abundance or transcriptional activity of YAP (as
assessed by gene expression signatures [18]) be used in the clinic for
classification of sarcomas and prediction of clinical outcome?
– Does the Hippo pathway mostly regulate generic functions such as
proliferation and apoptosis or does it consistently interfere with
lineage-specific fate determination gene expression as in ERMS
where it mainly suppresses the expression of differentiated muscle
genes [18]?
– Why is there a nearly 100% penetrance of WWTR1-CAMTA or YAP1-
TFE3 fusion genes in epithelioid haemangioendothelioma? What
do these fusion genes do and why are they so important in this
type of sarcoma?
Even if the role of the Hippo pathway in cancer is now well
established, Hippo research is still in its infancy compared with other tu-
mour suppressor and oncogenic proteins and pathways such as P53,
MYC or oncogenic RAS, and there is still much left to understand before
achieving full control over this intriguing pathway to specifically target
sarcomas and other cancers.
127
Supplementary data to this article can be found online at http://dx.
doi.org/10.1016/j.bbcan.2015.05.006.
Transparency document
The Transparency document associated with this article can be
found, in the online version.
Funding
Work in the University of Aberdeen laboratories is supported by
grants from Sarcoma UK (reference 004.2012), Friends of Anchor (refer-
ences 14004 and 14005), and the Medical Research Council (grant num-
ber 99477). Annie M. Tremblay is recipient of a postdoctoral fellowship
from the Canadian Institutes of Health Research (CIHR).
References
[1] R.S. Udan, M. Kango-Singh, R. Nolo, C. Tao, G. Halder, Hippo promotes proliferation ar-
rest and apoptosis in the Salvador/Warts pathway, Nat. Cell Biol. 5 (2003) 914–920.
[2] K.F. Harvey, X. Zhang, D.M. Thomas, The Hippo pathway and human cancer, Nat.
Rev. Cancer 13 (2013) 246–257.
[3] T. Moroishi, C.G. Hansen, K.L. Guan, The emerging roles of YAP and TAZ in cancer,
nature reviews, Cancer 15 (2015) 73–79.
[4] K. Harvey, N. Tapon, The Salvador–Warts–Hippo pathway — an emerging tumour-
suppressor network, Nat. Rev. Cancer 7 (2007) 182–191.
[5] D. Pan, The hippo signaling pathway in development and cancer, Dev. Cell 19
(2010) 491–505.
[6] B.S. Taylor, J. Barretina, R.G. Maki, C.R. Antonescu, S. Singer, M. Ladanyi, Advances in
sarcoma genomics and new therapeutic targets, nature reviews, Cancer 11 (2011)
541–557.
[7] J.R.F. Goldblum, A.L., S. Weiss, Enzinger and Weiss's Soft Tissue Tumors, 6th ed.
Elsevier, 2014.
[8] J.L. Anderson, C.T. Denny, W.D. Tap, N. Federman, Pediatric sarcomas: translating
molecular pathogenesis of disease to novel therapeutic possibilities, Pediatr. Res.
72 (2012) 112–121.
[9] B.S. Fletcher, J.A. Bridge, P.C.D. Hogendoorn, F. Mertens, World Health Organisation,
Classification of tumours: Tumours of Soft Tissue and Bone, 4th ed. WHO Press,
2013.
[10] C.D. Fletcher, The evolving classification of soft tissue tumours — an update based
on the new 2013 WHO classification, Histopathology 64 (2014) 2–11.
[11] C. Fisher, Immunohistochemistry in diagnosis of soft tissue tumours, Histopathol-
ogy 58 (2011) 1001–1012.
[12] C. Fisher, Dataset for Histopathology Reporting of Soft Tissue Sarcomas, in: T.R.C.o.
Pathologists (Ed.), 2014.
[13] M.S. Lawrence, P. Stojanov, C.H. Mermel, J.T. Robinson, L.A. Garraway, T.R. Golub,
M. Meyerson, S.B. Gabriel, E.S. Lander, G. Getz, Discovery and saturation analysis
of cancer genes across 21 tumour types, Nature 505 (2014) 495–501.
[14] B. Vogelstein, N. Papadopoulos, V.E. Velculescu, S. Zhou, L.A. Diaz Jr., K.W. Kinzler,
Cancer genome landscapes, Science 339 (2013) 1546–1558.
[15] C.R. Antonescu, F. Le Loarer, J.M. Mosquera, A. Sboner, L. Zhang, C.L. Chen, H.W.
Chen, N. Pathan, T. Krausz, B.C. Dickson, I. Weinreb, M.A. Rubin, M. Hameed, C.D.
Fletcher, Novel YAP1-TFE3 fusion defines a distinct subset of epithelioid
hemangioendothelioma, Genes Chromosom. Cancer 52 (2013) 775–784.
[16] C. Errani, L. Zhang, Y.S. Sung, M. Hajdu, S. Singer, R.G. Maki, J.H. Healey, C.R.
Antonescu, A novel WWTR1-CAMTA1 gene fusion is a consistent abnormality in
epithelioid hemangioendothelioma of different anatomic sites, Genes Chromosom.
Cancer 50 (2011) 644–653.
[17] Z. Helias-Rodzewicz, G. Perot, F. Chibon, C. Ferreira, P. Lagarde, P. Terrier, J.M.
Coindre, A. Aurias, YAP1 and VGLL3, encoding two cofactors of TEAD transcription
factors, are amplified and overexpressed in a subset of soft tissue sarcomas, Genes
Chromosom. Cancer 49 (2010) 1161–1171.
[18] A.M. Tremblay, E. Missiaglia, G.G. Galli, S. Hettmer, R. Urcia, M. Carrara, R.N. Judson,
K. Thway, G. Nadal, J.L. Selfe, G. Murray, R.A. Calogero, C. De Bari, P.S. Zammit, M.
Delorenzi, A.J. Wagers, J. Shipley, H. Wackerhage, F.D. Camargo, The Hippo trans-
ducer YAP1 transforms activated satellite cells and is a potent effector of embryo-
nal rhabdomyosarcoma formation, Cancer Cell 26 (2014) 273–287.
[19] K.H. Hallor, R. Sciot, J. Staaf, M. Heidenblad, A. Rydholm, H.C. Bauer, K. Astrom, H.A.
Domanski, J.M. Meis, L.G. Kindblom, I. Panagopoulos, N. Mandahl, F. Mertens, Two
genetic pathways, t(1;10) and amplification of 3p11-12, in myxoinflammatory fi-
broblastic sarcoma, haemosiderotic fibrolipomatous tumour, and morphologically
similar lesions, J. Pathol. 217 (2009) 716–727.
[20] K. Gambaro, M.C. Quinn, P.M. Wojnarowicz, S.L. Arcand, M. de Ladurantaye, V.
Barres, J.S. Ripeau, A.M. Killary, E.C. Davis, J. Lavoie, D.M. Provencher, A.M. Mes-
Masson, M. Chevrette, P.N. Tonin, VGLL3 expression is associated with a tumor
suppressor phenotype in epithelial ovarian cancer, Mol. Oncol. 7 (2013) 513–530.
[21] D.M. Parham, D.A. Ellison, Rhabdomyosarcomas in adults and children: an update,
Arch. Pathol. Lab. Med. 130 (2006) 1454–1465.
[22] D. Van Mater, L. Ano, J.M. Blum, M.T. Webster, W. Huang, N. Williams, Y. Ma, D.M.
Cardona, C.M. Fan, D.G. Kirsch, Acute tissue injury activates satellite cells and
128 A.D. Mohamed et al. / Biochimica et Biophysica Acta 1856 (2015) 121–129
promotes sarcoma formation via the HGF/c-MET signaling pathway, Cancer Res. 75
(2015) 605–614.
[23] D.A. Liebner, The indications and efficacy of conventional chemotherapy in prima-
ry and recurrent sarcoma, J. Surg. Oncol. 111 (2015) 622–631.
[24] A. Gronchi, C. Colombo, C.P. Raut, Surgical management of localized soft tissue tu-
mors, Cancer 120 (2014) 2638–2648.
[25] M. Linch, A.B. Miah, K. Thway, I.R. Judson, C. Benson, Systemic treatment of soft-
tissue sarcoma-gold standard and novel therapies, nature reviews, Clin. Oncol. 11
(2014) 187–202.
[26] S.P. D'Angelo, W.D. Tap, G.K. Schwartz, R.D. Carvajal, Sarcoma immunotherapy:
past approaches and future directions, Sarcoma 2014 (2014) 391967.
[27] S. Radaelli, S. Stacchiotti, P.G. Casali, A. Gronchi, Emerging therapies for adult soft
tissue sarcoma, Expert. Rev. Anticancer. Ther. 14 (2014) 689–704.
[28] T. Al-Zaid, N. Somaiah, A.J. Lazar, Targeted therapies for sarcomas: new roles for the
pathologist, Histopathology 64 (2014) 119–133.
[29] C. Forscher, M. Mita, R. Figlin, Targeted therapy for sarcomas, Biologics Targets
Ther. 8 (2014) 91–105.
[30] A. Neuville, F. Chibon, J.M. Coindre, Grading of soft tissue sarcomas: from histolog-
ical to molecular assessment, Pathology 46 (2014) 113–120.
[31] S.B. Edge, D.R., C.C. Compton, A.G. Fritz, F.L. Greene, A. Trotti, AJCC Cancer Staging
Manual, 7th ed. Springer, 2010.
[32] E. Gateff, Malignant neoplasms of genetic origin in Drosophila melanogaster, Sci-
ence 200 (1978) 1448–1459.
[33] J. Avruch, D. Zhou, J. Fitamant, N. Bardeesy, F. Mou, L.R. Barrufet, Protein kinases of the
Hippo pathway: regulation and substrates, Semin. Cell Dev. Biol. 23 (2012) 770–784.
[34] T. Oka, V. Mazack, M. Sudol, Mst2 and Lats kinases regulate apoptotic function of
Yes kinase-associated protein (YAP), J. Biol. Chem. 283 (2008) 27534–27546.
[35] B. Zhao, L. Li, K. Tumaneng, C.Y. Wang, K.L. Guan, A coordinated phosphorylation by
Lats and CK1 regulates YAP stability through SCF(beta-TRCP), Genes Dev. 24
(2010) 72–85.
[36] Q.Y. Lei, H. Zhang, B. Zhao, Z.Y. Zha, F. Bai, X.H. Pei, S. Zhao, Y. Xiong, K.L. Guan, TAZ
promotes cell proliferation and epithelial-mesenchymal transition and is inhibited
by the hippo pathway, Mol. Cell. Biol. 28 (2008) 2426–2436.
[37] J. Rosenbluh, D. Nijhawan, A.G. Cox, X. Li, J.T. Neal, E.J. Schafer, T.I. Zack, X. Wang, A.
Tsherniak, A.C. Schinzel, D.D. Shao, S.E. Schumacher, B.A. Weir, F. Vazquez, G.S.
Cowley, D.E. Root, J.P. Mesirov, R. Beroukhim, C.J. Kuo, W. Goessling, W.C. Hahn,
beta-Catenin-driven cancers require a YAP1 transcriptional complex for survival
and tumorigenesis, Cell 151 (2012) 1457–1473.
[38] M. Sudol, Yes-associated protein (YAP65) is a proline-rich phosphoprotein that
binds to the SH3 domain of the Yes proto-oncogene product, Oncogene 9 (1994)
2145–2152.
[39] M. Sudol, P. Bork, A. Einbond, K. Kastury, T. Druck, M. Negrini, K. Huebner, D.
Lehman, Characterization of the mammalian YAP (Yes-associated protein) gene
and its role in defining a novel protein module, the WW domain, J. Biol. Chem.
270 (1995) 14733–14741.
[40] B. Zhao, X. Ye, J. Yu, L. Li, W. Li, S. Li, J. Yu, J.D. Lin, C.Y. Wang, A.M. Chinnaiyan, Z.C.
Lai, K.L. Guan, TEAD mediates YAP-dependent gene induction and growth control,
Genes Dev. 22 (2008) 1962–1971.
[41] L.M. Koontz, Y. Liu-Chittenden, F. Yin, Y. Zheng, J. Yu, B. Huang, Q. Chen, S. Wu, D.
Pan, The Hippo effector yorkie controls normal tissue growth by antagonizing
scalloped-mediated default repression, Dev. Cell 25 (2013) 388–401.
[42] E. Aragon, N. Goerner, A.I. Zaromytidou, Q. Xi, A. Escobedo, J. Massague, M.J. Macias,
A Smad action turnover switch operated by WW domain readers of a
phosphoserine code, Genes Dev. 25 (2011) 1275–1288.
[43] M. Murakami, M. Nakagawa, E.N. Olson, O. Nakagawa, A WW domain protein TAZ
is a critical coactivator for TBX5, a transcription factor implicated in Holt–Oram
syndrome, Proc. Natl. Acad. Sci. U. S. A. 102 (2005) 18034–18039.
[44] S. Dupont, L. Morsut, M. Aragona, E. Enzo, S. Giulitti, M. Cordenonsi, F. Zanconato,
D.J. Le, M. Forcato, S. Bicciato, N. Elvassore, S. Piccolo, Role of YAP/TAZ in
mechanotransduction, Nature 474 (2011) 179–183.
[45] L. Azzolin, F. Zanconato, S. Bresolin, M. Forcato, G. Basso, S. Bicciato, M. Cordenonsi,
S. Piccolo, Role of TAZ as mediator of Wnt signaling, Cell 151 (2012) 1443–1456.
[46] L. Azzolin, T. Panciera, S. Soligo, E. Enzo, S. Bicciato, S. Dupont, S. Bresolin, C.
Frasson, G. Basso, V. Guzzardo, A. Fassina, M. Cordenonsi, S. Piccolo, YAP/TAZ incor-
poration in the beta-catenin destruction complex orchestrates the Wnt response,
Cell 158 (2014) 157–170.
[47] F.X. Yu, B. Zhao, N. Panupinthu, J.L. Jewell, I. Lian, L.H. Wang, J. Zhao, H. Yuan, K.
Tumaneng, H. Li, X.D. Fu, G.B. Mills, K.L. Guan, Regulation of the Hippo-YAP path-
way by G-protein-coupled receptor signaling, Cell 150 (2012) 780–791.
[48] J. Dong, G. Feldmann, J. Huang, S. Wu, N. Zhang, S.A. Comerford, M.F. Gayyed, R.A.
Anders, A. Maitra, D. Pan, Elucidation of a universal size-control mechanism in Dro-
sophila and mammals, Cell 130 (2007) 1120–1133.
[49] F.D. Camargo, S. Gokhale, J.B. Johnnidis, D. Fu, G.W. Bell, R. Jaenisch, T.R.
Brummelkamp, YAP1 increases organ size and expands undifferentiated progeni-
tor cells, Curr. Biol. 17 (2007) 2054–2060.
[50] K. Schlegelmilch, M. Mohseni, O. Kirak, J. Pruszak, J.R. Rodriguez, D. Zhou, B.T.
Kreger, V. Vasioukhin, J. Avruch, T.R. Brummelkamp, F.D. Camargo, Yap1 acts
downstream of alpha-catenin to control epidermal proliferation, Cell 144 (2011)
782–795.
[51] T. Yu, J. Bachman, Z.C. Lai, Mutation analysis of large tumor suppressor genes LATS1
and LATS2 supports a tumor suppressor role in human cancer, Protein Cell 6
(2015) 6–11.
[52] J. Gao, B.A. Aksoy, U. Dogrusoz, G. Dresdner, B. Gross, S.O. Sumer, Y. Sun, A.
Jacobsen, R. Sinha, E. Larsson, E. Cerami, C. Sander, N. Schultz, Integrative analysis
of complex cancer genomics and clinical profiles using the cBioPortal, Sci. Signal. 6
(2013) l1.
[53] E. Cerami, J. Gao, U. Dogrusoz, B.E. Gross, S.O. Sumer, B.A. Aksoy, A. Jacobsen, C.J.
Byrne, M.L. Heuer, E. Larsson, Y. Antipin, B. Reva, A.P. Goldberg, C. Sander, N.
Schultz, The cBio cancer genomics portal: an open platform for exploring multidi-
mensional cancer genomics data, Cancer Discov. 2 (2012) 401–404.
[54] M.S. Lawrence, P. Stojanov, P. Polak, G.V. Kryukov, K. Cibulskis, A. Sivachenko, S.L.
Carter, C. Stewart, C.H. Mermel, S.A. Roberts, A. Kiezun, P.S. Hammerman, A.
McKenna, Y. Drier, L. Zou, A.H. Ramos, T.J. Pugh, N. Stransky, E. Helman, J. Kim, C.
Sougnez, L. Ambrogio, E. Nickerson, E. Shefler, M.L. Cortes, D. Auclair, G. Saksena, D.
Voet, M. Noble, D. DiCara, P. Lin, L. Lichtenstein, D.I. Heiman, T. Fennell, M.
Imielinski, B. Hernandez, E. Hodis, S. Baca, A.M. Dulak, J. Lohr, D.A. Landau, C.J. Wu, J.
Melendez-Zajgla, A. Hidalgo-Miranda, A. Koren, S.A. McCarroll, J. Mora, R.S. Lee, B.
Crompton, R. Onofrio, M. Parkin, W. Winckler, K. Ardlie, S.B. Gabriel, C.W. Roberts,
J.A. Biegel, K. Stegmaier, A.J. Bass, L.A. Garraway, M. Meyerson, T.R. Golub, D.A.
Gordenin, S. Sunyaev, E.S. Lander, G. Getz, Mutational heterogeneity in cancer and
the search for new cancer-associated genes, Nature 499 (2013) 214–218.
[55] J. Sima, D.M. Gilbert, Complex correlations: replication timing and mutational land-
scapes during cancer and genome evolution, Curr. Opin. Genet. Dev. 25C (2014)
93–100.
[56] M. Overholtzer, J. Zhang, G.A. Smolen, B. Muir, W. Li, D.C. Sgroi, C.X. Deng, J.S.
Brugge, D.A. Haber, Transforming properties of YAP, a candidate oncogene on
the chromosome 11q22 amplicon, Proc. Natl. Acad. Sci. U. S. A. 103 (2006)
12405–12410.
[57] D.D. Shao, W. Xue, E.B. Krall, A. Bhutkar, F. Piccioni, X. Wang, A.C. Schinzel, S. Sood,
J. Rosenbluh, J.W. Kim, Y. Zwang, T.M. Roberts, D.E. Root, T. Jacks, W.C. Hahn, KRAS
and YAP1 converge to regulate EMT and tumor survival, Cell 158 (2014) 171–184.
[58] A. Kapoor, W. Yao, H. Ying, S. Hua, A. Liewen, Q. Wang, Y. Zhong, C.J. Wu, A.
Sadanandam, B. Hu, Q. Chang, G.C. Chu, R. Al-Khalil, S. Jiang, H. Xia, E. Fletcher-
Sananikone, C. Lim, G.I. Horwitz, A. Viale, P. Pettazzoni, N. Sanchez, H. Wang, A.
Protopopov, J. Zhang, T. Heffernan, R.L. Johnson, L. Chin, Y.A. Wang, G. Draetta,
R.A. DePinho, Yap1 activation enables bypass of oncogenic kras addiction in pan-
creatic cancer, Cell 158 (2014) 185–197.
[59] W. Zhang, N. Nandakumar, Y. Shi, M. Manzano, A. Smith, G. Graham, S. Gupta, E.E.
Vietsch, S.Z. Laughlin, M. Wadhwa, M. Chetram, M. Joshi, F. Wang, B. Kallakury, J.
Toretsky, A. Wellstein, C. Yi, Downstream of mutant KRAS, the transcription regu-
lator YAP is essential for neoplastic progression to pancreatic ductal adenocarcino-
ma, Sci. Signal. 7 (2014) ra42.
[60] M.A. St John, W. Tao, X. Fei, R. Fukumoto, M.L. Carcangiu, D.G. Brownstein, A.F.
Parlow, J. McGrath, T. Xu, Mice deficient of Lats1 develop soft-tissue sarcomas,
ovarian tumours and pituitary dysfunction, Nat. Genet. 21 (1999) 182–186.
[61] A.I. McClatchey, I. Saotome, K. Mercer, D. Crowley, J.F. Gusella, R.T. Bronson, T. Jacks,
Mice heterozygous for a mutation at the Nf2 tumor suppressor locus develop a
range of highly metastatic tumors, Genes Dev. 12 (1998) 1121–1133.
[62] M. Nishio, K. Hamada, K. Kawahara, M. Sasaki, F. Noguchi, S. Chiba, K. Mizuno, S.O.
Suzuki, Y. Dong, M. Tokuda, T. Morikawa, H. Hikasa, J. Eggenschwiler, N. Yabuta, H.
Nojima, K. Nakagawa, Y. Hata, H. Nishina, K. Mimori, M. Mori, T. Sasaki, T.W. Mak,
T. Nakano, S. Itami, A. Suzuki, Cancer susceptibility and embryonic lethality in
Mob1a/1b double-mutant mice, J. Clin. Invest. 122 (2012) 4505–4518.
[63] F. Hamaratoglu, M. Willecke, M. Kango-Singh, R. Nolo, E. Hyun, C. Tao, H. Jafar-
Nejad, G. Halder, The tumour-suppressor genes NF2/merlin and expanded act
through Hippo signalling to regulate cell proliferation and apoptosis, Nat. Cell
Biol. 8 (2006) 27–36.
[64] N. Zhang, H. Bai, K.K. David, J. Dong, Y. Zheng, J. Cai, M. Giovannini, P. Liu, R.A.
Anders, D. Pan, The Merlin/NF2 tumor suppressor functions through the YAP
oncoprotein to regulate tissue homeostasis in mammals, Dev. Cell 19 (2010)
27–38.
[65] A. Hergovich, MOB control: reviewing a conserved family of kinase regulators, Cell.
Signal. 23 (2011) 1433–1440.
[66] K.I. Watt, R. Judson, P. Medlow, K. Reid, T.B. Kurth, J.G. Burniston, A. Ratkevicius, C.
De Bari, H. Wackerhage, Yap is a novel regulator of C2C12 myogenesis, Biochem.
Biophys. Res. Commun. 393 (2010) 619–624.
[67] R.N. Judson, A.M. Tremblay, P. Knopp, R.B. White, R. Urcia, B.C. De, P.S. Zammit, F.D.
Camargo, H. Wackerhage, The Hippo pathway member Yap plays a key role in
influencing fate decisions in muscle satellite cells, J. Cell Sci. 125 (2012)
6009–6019.
[68] G. Pallafacchina, S. Francois, B. Regnault, B. Czarny, V. Dive, A. Cumano, D.
Montarras, M. Buckingham, An adult tissue-specific stem cell in its niche: a gene
profiling analysis of in vivo quiescent and activated muscle satellite cells, Stem
Cell Res. 4 (2010) 77–91.
[69] J.S. Chamberlain, J. Metzger, M. Reyes, D. Townsend, J.A. Faulkner, Dystrophin-
deficient mdx mice display a reduced life span and are susceptible to spontaneous
rhabdomyosarcoma, FASEB J. 21 (2007) 2195–2204.
[70] V. Hosur, A. Kavirayani, J. Riefler, L.M. Carney, B. Lyons, B. Gott, G.A. Cox, L.D. Shultz,
Dystrophin and dysferlin double mutant mice: a novel model for rhabdomyosarco-
ma, Cancer Genet. 205 (2012) 232–241.
[71] U. Flucke, R.J. Vogels, N. de Saint Aubain Somerhausen, D.H. Creytens, R.G. Riedl,
J.M. van Gorp, A.N. Milne, C.J. Huysentruyt, M.A. Verdijk, M.M. van Asseldonk, A.J.
Suurmeijer, J. Bras, G. Palmedo, P.J. Groenen, T. Mentzel, Epithelioid
hemangioendothelioma: clinicopathologic, immunhistochemical, and molecular
genetic analysis of 39 cases, Diagn. Pathol. 9 (2014) 131.
[72] E. Missiaglia, D. Williamson, J. Chisholm, P. Wirapati, G. Pierron, F. Petel, J.P.
Concordet, K. Thway, O. Oberlin, K. Pritchard-Jones, O. Delattre, M. Delorenzi, J.
Shipley, PAX3/FOXO1 fusion gene status is the key prognostic molecular marker
in rhabdomyosarcoma and significantly improves current risk stratification, J.
Clin. Oncol. 30 (2012) 1670–1677.
[73] J.F. Shern, L. Chen, J. Chmielecki, J.S. Wei, R. Patidar, M. Rosenberg, L. Ambrogio, D.
Auclair, J. Wang, Y.K. Song, C. Tolman, L. Hurd, H. Liao, S. Zhang, D. Bogen, A.S.
 A.D. Mohamed et al. / Biochimica et Biophysica Acta 1856 (2015) 121–129
Brohl, S. Sindiri, D. Catchpoole, T. Badgett, G. Getz, J. Mora, J.R. Anderson, S.X.
Skapek, F.G. Barr, M. Meyerson, D.S. Hawkins, J. Khan, Comprehensive genomic
analysis of rhabdomyosarcoma reveals a landscape of alterations affecting a com-
mon genetic axis in fusion-positive and fusion-negative tumors, Cancer Discov. 4
(2014) 216–231.
[74] J.M. Mosquera, A. Sboner, L. Zhang, N. Kitabayashi, C.L. Chen, Y.S. Sung, L.H. Wexler,
M.P. LaQuaglia, M. Edelman, C. Sreekantaiah, M.A. Rubin, C.R. Antonescu, Recurrent
NCOA2 gene rearrangements in congenital/infantile spindle cell rhabdomyosarco-
ma, Genes Chromosom. Cancer 52 (2013) 538–550.
[75] M.R. Tanas, S. Ma, F.O. Jadaan, C.K. Ng, B. Weigelt, J.S. Reis-Filho, B.P. Rubin, Mech-
anism of action of a WWTR1(TAZ)-CAMTA1 fusion oncoprotein, Oncogene (2015)
(in press).
[76] G. Ciriello, M.L. Miller, B.A. Aksoy, Y. Senbabaoglu, N. Schultz, C. Sander, Emerging
landscape of oncogenic signatures across human cancers, Nat. Genet. 45 (2013)
1127–1133.
[77] J. Barretina, B.S. Taylor, S. Banerji, A.H. Ramos, M. Lagos-Quintana, P.L. Decarolis, K.
Shah, N.D. Socci, B.A. Weir, A. Ho, D.Y. Chiang, B. Reva, C.H. Mermel, G. Getz, Y.
Antipin, R. Beroukhim, J.E. Major, C. Hatton, R. Nicoletti, M. Hanna, T. Sharpe, T.J.
Fennell, K. Cibulskis, R.C. Onofrio, T. Saito, N. Shukla, C. Lau, S. Nelander, S.J.
Silver, C. Sougnez, A. Viale, W. Winckler, R.G. Maki, L.A. Garraway, A. Lash, H.
Greulich, D.E. Root, W.R. Sellers, G.K. Schwartz, C.R. Antonescu, E.S. Lander, H.E.
Varmus, M. Ladanyi, C. Sander, M. Meyerson, S. Singer, Subtype-specific genomic
alterations define new targets for soft-tissue sarcoma therapy, Nat. Genet. 42
(2010) 715–721.
[78] C.R. Antonescu, L. Zhang, G.P. Nielsen, A.E. Rosenberg, P. Dal Cin, C.D. Fletcher, Con-
sistent t(1;10) with rearrangements of TGFBR3 and MGEA5 in both
myxoinflammatory fibroblastic sarcoma and hemosiderotic fibrolipomatous
tumor, Genes Chromosom. Cancer 50 (2011) 757–764.
[79] S. Jiao, H. Wang, Z. Shi, A. Dong, W. Zhang, X. Song, F. He, Y. Wang, Z. Zhang, W.
Wang, X. Wang, T. Guo, P. Li, Y. Zhao, H. Ji, L. Zhang, Z. Zhou, A peptide mimicking
VGLL4 function acts as a YAP antagonist therapy against gastric cancer, Cancer Cell
25 (2014) 166–180.
[80] Y. Takahashi, Y. Miyoshi, C. Takahata, N. Irahara, T. Taguchi, Y. Tamaki, S. Noguchi,
Down-regulation of LATS1 and LATS2 mRNA expression by promoter hypermethy-
lation and its association with biologically aggressive phenotype in human breast
cancers, Clin. Cancer Res. 11 (2005) 1380–1385.
[81] Z. Jiang, X. Li, J. Hu, W. Zhou, Y. Jiang, G. Li, D. Lu, Promoter hypermethylation-
mediated down-regulation of LATS1 and LATS2 in human astrocytoma, Neurosci.
Res. 56 (2006) 450–458.
[82] C. Seidel, U. Schagdarsurengin, K. Blumke, P. Wurl, G.P. Pfeifer, S. Hauptmann, H.
Taubert, R. Dammann, Frequent hypermethylation of MST1 and MST2 in soft tissue
sarcoma, Mol. Carcinog. 46 (2007) 865–871.
[83] A.M. Tremblay, F.D. Camargo, Hippo signaling in mammalian stem cells, Semin.
Cell Dev. Biol. 23 (2012) 818–826.
[84] L.E. Crose, K.A. Galindo, J.G. Kephart, C. Chen, J. Fitamant, N. Bardeesy, R.C. Bentley,
R.L. Galindo, J.T. Ashley Chi, C.M. Linardic, Alveolar rhabdomyosarcoma-associated
PAX3-FOXO1 promotes tumorigenesis via Hippo pathway suppression, J. Clin. In-
vest. 124 (2014) 285–296.
[85] D. Williamson, E. Missiaglia, R.A. de, G. Pierron, B. Thuille, G. Palenzuela, K. Thway,
D. Orbach, M. Lae, P. Freneaux, K. Pritchard-Jones, O. Oberlin, J. Shipley, O. Delattre,
Fusion gene-negative alveolar rhabdomyosarcoma is clinically and molecularly in-
distinguishable from embryonal rhabdomyosarcoma, J. Clin. Oncol. 28 (2010)
2151–2158.
[86] X. Chen, E. Stewart, A.A. Shelat, C. Qu, A. Bahrami, M. Hatley, G. Wu, C. Bradley, J.
McEvoy, A. Pappo, S. Spunt, M.B. Valentine, V. Valentine, F. Krafcik, W.H. Lang, M.
Wierdl, L. Tsurkan, V. Tolleman, S.M. Federico, C. Morton, C. Lu, L. Ding, J. Easton,
M. Rusch, P. Nagahawatte, J. Wang, M. Parker, L. Wei, E. Hedlund, D. Finkelstein,
M. Edmonson, S. Shurtleff, K. Boggs, H. Mulder, D. Yergeau, S. Skapek, D.S.
Hawkins, N. Ramirez, P.M. Potter, J.A. Sandoval, A.M. Davidoff, E.R. Mardis, R.K.
Wilson, J. Zhang, J.R. Downing, M.A. Dyer, P. St, Jude Children's Research
Hospital-Washington University Pediatric Cancer Genome, targeting oxidative
stress in embryonal rhabdomyosarcoma, Cancer Cell 24 (2013) 710–724.
[87] L. Lin, A.J. Sabnis, E. Chan, V. Olivas, L. Cade, E. Pazarentzos, S. Asthana, D. Neel, J.J.
Yan, X. Lu, L. Pham, M.M. Wang, N. Karachaliou, M.G. Cao, J.L. Manzano, J.L.
Ramirez, J.M. Torres, F. Buttitta, C.M. Rudin, E.A. Collisson, A. Algazi, E. Robinson,
I. Osman, E. Munoz-Couselo, J. Cortes, D.T. Frederick, Z.A. Cooper, M. McMahon,
A. Marchetti, R. Rosell, K.T. Flaherty, J.A. Wargo, T.G. Bivona, The Hippo effector
YAP promotes resistance to RAF- and MEK-targeted cancer therapies, Nat. Genet.
47 (2015) 250–256.
129
[88] A.R. Hinson, R. Jones, L.E. Crose, B.C. Belyea, F.G. Barr, C.M. Linardic, Human rhabdo-
myosarcoma cell lines for rhabdomyosarcoma research: utility and pitfalls, Front.
Oncol. 3 (2013) 183.
[89] R. Fan, N.G. Kim, B.M. Gumbiner, Regulation of Hippo pathway by mitogenic
growth factors via phosphoinositide 3-kinase and phosphoinositide-dependent
kinase-1, Proc. Natl. Acad. Sci. U. S. A. 110 (2013) 2569–2574.
[90] Z. Yuan, D. Kim, S. Shu, J. Wu, J. Guo, L. Xiao, S. Kaneko, D. Coppola, J.Q. Cheng,
Phosphoinositide 3-kinase/Akt inhibits MST1-mediated pro-apoptotic signaling
through phosphorylation of threonine 120, J. Biol. Chem. 285 (2010) 3815–3824.
[91] S.W. Jang, S.J. Yang, S. Srinivasan, K. Ye, Akt phosphorylates MstI and prevents its
proteolytic activation, blocking FOXO3 phosphorylation and nuclear translocation,
J. Biol. Chem. 282 (2007) 30836–30844.
[92] F.K. Collak, K. Yagiz, D.J. Luthringer, B. Erkaya, B. Cinar, Threonine-120 phosphory-
lation regulated by phosphoinositide-3-kinase/Akt and mammalian target of
rapamycin pathway signaling limits the antitumor activity of mammalian sterile
20-like kinase 1, J. Biol. Chem. 287 (2012) 23698–23709.
[93] C.L. Yeung, V.N. Ngo, P.J. Grohar, F.I. Arnaldez, A. Asante, X. Wan, J. Khan, S.M.
Hewitt, C. Khanna, L.M. Staudt, L.J. Helman, Loss-of-function screen in rhabdomyo-
sarcoma identifies CRKL-YES as a critical signal for tumor growth, Oncogene 32
(2013) 5429–5438.
[94] C.Y. Liu, Z.Y. Zha, X. Zhou, H. Zhang, W. Huang, D. Zhao, T. Li, S.W. Chan, C.J. Lim, W.
Hong, S. Zhao, Y. Xiong, Q.Y. Lei, K.L. Guan, The hippo tumor pathway promotes
TAZ degradation by phosphorylating a phosphodegron and recruiting the
SCF{beta}-TrCP E3 ligase, J. Biol. Chem. 285 (2010) 37159–37169.
[95] K. Tu, W. Yang, C. Li, X. Zheng, Z. Lu, C. Guo, Y. Yao, Q. Liu, Fbxw7 is an independent
prognostic marker and induces apoptosis and growth arrest by regulating YAP
abundance in hepatocellular carcinoma, Mol. Cancer 13 (2014) 110.
[96] K. Brodowska, A. Al-Moujahed, A. Marmalidou, M. Meyer Zu Horste, J. Cichy, J.W.
Miller, E. Gragoudas, D.G. Vavvas, The clinically used photosensitizer Verteporfin
(VP) inhibits YAP-TEAD and human retinoblastoma cell growth in vitro without
light activation, Exp. Eye Res. 124 (2014) 67–73.
[97] Y. Liu-Chittenden, B. Huang, J.S. Shim, Q. Chen, S.J. Lee, R.A. Anders, J.O. Liu, D. Pan,
Genetic and pharmacological disruption of the TEAD-YAP complex suppresses the
oncogenic activity of YAP, Genes Dev. 26 (2012) 1300–1305.
[98] F.X. Yu, J. Luo, J.S. Mo, G. Liu, Y.C. Kim, Z. Meng, L. Zhao, G. Peyman, H. Ouyang, W.
Jiang, J. Zhao, X. Chen, L. Zhang, C.Y. Wang, B.C. Bastian, K. Zhang, K.L. Guan, Mutant
Gq/11 promote uveal melanoma tumorigenesis by activating YAP, Cancer Cell 25
(2014) 822–830.
[99] X. Feng, M.S. Degese, R. Iglesias-Bartolome, J.P. Vaque, A.A. Molinolo, M. Rodrigues,
M.R. Zaidi, B.R. Ksander, G. Merlino, A. Sodhi, Q. Chen, J.S. Gutkind, Hippo-
independent activation of YAP by the GNAQ uveal melanoma oncogene through
a trio-regulated rho GTPase signaling circuitry, Cancer Cell 25 (2014) 831–845.
[100] M. O'Hayre, M.S. Degese, J.S. Gutkind, Novel insights into G protein and G protein-
coupled receptor signaling in cancer, Curr. Opin. Cell Biol. 27 (2014) 126–135.
[101] J.W. Clendening, A. Pandyra, P.C. Boutros, S. El Ghamrasni, F. Khosravi, G.A. Trentin,
A. Martirosyan, A. Hakem, R. Hakem, I. Jurisica, L.Z. Penn, Dysregulation of the
mevalonate pathway promotes transformation, Proc. Natl. Acad. Sci. U. S. A. 107
(2010) 15051–15056.
[102] G. Sorrentino, N. Ruggeri, V. Specchia, M. Cordenonsi, M. Mano, S. Dupont, A.
Manfrin, E. Ingallina, R. Sommaggio, S. Piazza, A. Rosato, S. Piccolo, G. Del Sal, Met-
abolic control of YAP and TAZ by the mevalonate pathway, Nat. Cell Biol. 16 (2014)
357–366.
[103] Z. Wang, Y. Wu, H. Wang, Y. Zhang, L. Mei, X. Fang, X. Zhang, F. Zhang, H. Chen, Y.
Liu, Y. Jiang, S. Sun, Y. Zheng, N. Li, L. Huang, Interplay of mevalonate and Hippo
pathways regulates RHAMM transcription via YAP to modulate breast cancer cell
motility, Proc. Natl. Acad. Sci. U. S. A. 111 (2014) E89–E98.
[104] A.K. Altwairgi, Statins are potential anticancerous agents (Review), Oncol. Rep. 33
(2015) 1019–1039.
[105] A.V. Pobbati, S.W. Chan, I. Lee, H. Song, W. Hong, Structural and functional
similarity between the Vgll1-TEAD and the YAP-TEAD complexes, Structure 20
(2012) 1135–1140.
[106] Z. Zhou, T. Hu, Z. Xu, Z. Lin, Z. Zhang, T. Feng, L. Zhu, Y. Rong, H. Shen, J.M. Luk, X.
Zhang, N. Qin, Targeting Hippo pathway by specific interruption of YAP-TEAD in-
teraction using cyclic YAP-like peptides, FASEB J. 29 (2015) 724–732.
[107] R. Johnson, G. Halder, The two faces of Hippo: targeting the Hippo pathway for re-
generative medicine and cancer treatment, Nat. Rev. Drug Discov. 13 (2014) 63–79.