DISEASE IN WILDLIFE OR EXOTIC SPECIES
Epi helialeMesenchymal T ansi ion in a Case o
Me as a ic Thy oid Ca cinoma in a B own Bea
(U sus a c os)
Q4
Q3 A. Rod ı
´guez-La go
*
,R. de Miguel
*
,J. Ası
´n
*
,F. Choc eau
†
,M. Gimeno
*
,
P. Pinczowski
*
,A. de Ma ino
‡
and L. Lujan
*
*Depa men o Animal Pa hology, Uni e si y o Za agoza, Za agoza, Spain,
†
Laboni is, ONIRIS, Ecole Na ionale
Ve e inai e, Ag oalimen ai e e de l’Alimen a ion de Nan es-A lan ique, Nan es, F ance and
‡
Ins i u o A agones de Ciencias de
la Salud, Za agoza, Spain
Summa y
A 20-yea -old male b own bea (U sus a c os) wi h a 20 25 cm nec o ic mass adjacen o he achea was diag-
nosed as ha ing an anaplas ic hy oid ca cinoma. Me as ases we e obse ed in he lungs and one ad enal gland
and, his ologically, hese had anaplas ic and ollicula ca cinoma pa e ns, espec i ely. E-cadhe in labelling
was obse ed in he ad enal mass only, while N-cadhe in immunolabelling was de ec ed in he hy oid gland
and lung masses. Thy oid-speci ic ma ke s ( hy oid ansc ip ion ac o -1, hy oglobulin) we e exp essed in
he ad enal gland me as asis. This case illus a es an example o a p ima y epi helialemesenchymal ansi ion
enabling me as asis o dis an o gan si es, ollowed by a mesenchymaleepi helial ansi ion wi hin he ad enal
gland mic oen i onmen , allowing in asion and eacquisi ion o hy oid epi helial cell ea u es.
Epi helialemesenchymal ansi ions help o unde s and he phenomenon o ca cinoma cell plas ici y in
enabling coloniza ion and g ow h o me as ases.
Ó2020 Else ie L d. All igh s ese ed.
Keywo ds: b own bea ; epi helialemesenchymal ansi ion; hy oid ca cinoma
Anaplas ic hy oid ca cinomas a e one o he mos
agg essi e solid umou s in dogs (Lip ak, 2007) and
man, bu hey ep esen only 1e2% o all hy oid
neoplasms (Molina o e al., 2017). Thy oid neoplasms
end o in ade he bloods eam and ha e high a es o
me as asis (Lip ak, 2007). Thy oid gland ca cinomas
ha e been desc ibed in domes ic and wild animals
(Rosol and G €one, 2016). A ew cases o endoc ine
neoplasms ha e been desc ibed in he U sidae amily
(Lomba d and Wi e, 1959; Hubba d e al., 1983).
Immunohis ochemical cha ac e iza ion o hy oid
neoplasms includes use o ma ke s such as hy oid
ansc ip ion ac o (TTF)-1, a hy oid-speci ic p o-
ein, he exp ession o which con i ms a hy oid o igin
(Ramos-Va a e al., 2016); hy oglobulin (TGB), a
hy oid ho mone used o classi y hy oid neoplasms
(Ramos-Va a e al., 2016); and cy oke a in and E-
cadhe in (Fadda and Rossi, 2014). E-cadhe in is an
adhesion p o ein wi h an impo an ole in he o ma-
ion o epi helial cell- o-cell junc ions (Onde e al.,
2008). Down egula ion o E-cadhe in is conside ed
he c ucial e en in dedi e en ia ion, p og ession
and me as asis o ca cinomas (Liu and Lin, 2015).
Mesenchymal ma ke s such as imen in and N-cad-
he in can be used o complemen he in o ma ion p o-
ided by hese an ibodies (Onde e al., 2008).
Epi helialemesenchymal ansi ion (EMT) is a
cellula p ocess cha ac e ized by he con e sion o
epi helial cells o a mesenchymal pheno ype. EMT
plays a physiological ole du ing emb yogenesis
(Lee e al., 2005). This p ocess is silenced du ing
J. Comp. Pa h. xxxx, Vol. xxx, xxx A ailable online a www.sciencedi ec .com
ScienceDi ec
www.else ie .com/loca e/jcpa
Co espondence o: L. Lujan (e-mail: [email p o ec ed]).
0021-9975/$ - see on ma e Ó2020 Else ie L d. All igh s ese ed.
h ps://doi.o g/10.1016/j.jcpa.2020.01.004
Please ci e his a icle as: Rod ı´guez-La go A e al., Epi helialeMesenchymal T ansi ion in a Case o Me as a ic Thy oid Ca cinoma in a B own Bea
(U sus a c os), Jou nal o Compa a i e Pa hology, h ps://doi.o g/10.1016/j.jcpa.2020.01.004
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YJCPA2305_p oo ■14 Feb ua y 2020 ■1/4
adul hood, bu i can be eac i a ed by di e en pa h-
ological condi ions (Zida e al., 2018; Dong e and
Weinbe g, 2019). EMT allows p ima y neoplas ic
epi helial cells o acqui e he capaci y o mig a e
and o me as asize (Lee e al., 2005). In hese newly-
colonized issues, neoplas ic cells can unde go he
so-called mesenchymaleepi helial ansi ion
(MET), a e e se p ocess ha can occu in esponse
o di e en mic oen i onmen al signals. As a conse-
quence, me as ases can e-acqui e a di e en ia ed
epi helial pheno ype simila o ha o he no mal is-
sue whe e he me as a ic umou i s appea ed
(Thie y, 2002).
He ein we epo he deg ees o cellula di e en ia-
ion in pulmona y and ad enal gland me as ases
obse ed in a case o anaplas ic hy oid ca cinoma
in a b own bea (U sus a c os). These indings and
he immunohis ochemical ea u es, sugges ed an
EMT p ocess in he hy oid neoplas ic cells and a
MET p ocess in he ad enal gland me as ases.
A 20-yea -old male b own bea , bo n and aised a
a ci cus, was e e ed wi h an expanding, i m mass
adjacen o he achea. Dyspnoea, ano exia and p o-
g essi e weigh loss we e he majo clinical signs. Ra-
diog aphical examina ion o he lungs e ealed
mul iple pulmona y nodules and he animal was hu-
manely des oyed.
A nec opsy examina ion, an i egula , i m da k
b own, solid mass (20 25 cm) was ound in he hy-
oid egion. The cu su ace e ealed a la ge cen al
nec o ic a ea (app oxima ely 22 cm in diame e ),
wi h a 2 cm ing o pe iphe al iable issue. The lungs
exhibi ed mul iple nodules o di e en sizes
(0.5e15 cm), dis ibu ed andomly and associa ed
wi h an in ense medias inal p oli e a ion
(Supplemen a y Fig. 1). In he igh ad enal gland,
a pale yellow mass (3 5 cm) mass was p esen
(Supplemen a y Fig. 2). Tissue samples we e p o-
cessed ou inely and embedded in pa a in wax. Sec-
ions we e s ained by haema oxylin and eosin (HE)
and se ial sec ions we e subjec ed o immunohis o-
chemis y (IHC) (Supplemen a y Table 1).
Mic oscopically, he hy oid mass was encapsu-
la ed, highly cellula and had a b oad nec o ic cen e.
The neoplasm was composed o polyhed al o spindle-
shaped cells ei he in shee s o in in e lacing bundles,
s eams and who ls. The cells we e ill-de ined, and
showed a a iable amoun eosinophilic cy oplasm
(Supplemen a y Fig. 3). Nuclei a ied in size and
shape and had one o ou p ominen eosinophilic
nucleoli (Supplemen a y Fig. 4). Anisoka yosis and
anisocy osis we e ma ked and he mi o ic index was
high (11 mi oses pe 10 400 ields). No no mal hy-
oid issue was obse ed. Agg ega es o neoplas ic cells
we e equen ly obse ed in an in a ascula loca ion
(Fig. 1). Pulmona y nodules showed a his ological
pa e n simila o he hy oid mass, wi hou iden i i-
able no mal hy oid issue (Supplemen a y Figs. 5
and 6). The ad enal gland me as asis was o ganized
in a iably-sized ollicles wi h in aluminal eosino-
philic homogeneous ma e ial, esembling colloid
( hy oid ollicle-like s uc u es), some imes su -
ounded by solid a eas o polygonal, hy oid-like
neoplas ic cells. Anisoka yosis and anisocy osis we e
mild and he mi o ic index was low (1 mi osis pe
10 400 ields) (Supplemen a y Figs. 7 and 8).
Neoplas ic cells in he hy oid and lung masses we e
nega i e o E-cadhe in, bu had cy oplasmic exp es-
sion o N-cadhe in (Figs. 1 and 2;Supplemen a y
Figs. 9 and 10). Mo eo e , he neoplas ic cells ex-
hibi ed s ong cy oplasmic exp ession o imen in.
In con as , cells in he ad enal mass showed he
e e se pa e n o immunolabelling, ha ing s ong
cy oplasmic eac i i y o E-cadhe in, low signal o
imen in and no N-cadhe in exp ession (Figs. 3 and
4,Supplemen a y Table 2). The cy oplasm o he
neoplas ic cells in he hy oid, lung and ad enal
umou masses we e s ongly posi i e o pancy oke -
a in (Supplemen a y Figs. 11,13 and 15). TTF-1 and
TGB we e exp essed spa sely in he hy oid and lung
masses, bu he ad enal me as ases had low nuclea
exp ession o TTF-1 and s ong cy oplasmic eac-
i i y agains TGB (Supplemen a y Figs. 12,14 and
16).
Based on hese indings, an anaplas ic hy oid ca -
cinoma wi h me as ases o he lung and he ad enal
gland was diagnosed. Lung me as ases showed an un-
di e en ia ed, anaplas ic g ow h pa e n, simila o
he p oli e a ion obse ed in he hy oid gland. How-
e e , he ad enal mass exhibi ed well-di e en ia ed
cell p oli e a ion wi h ecognizable hy oid ollicle-
Fig. 1. Thy oid. Clus e o neoplas ic cells wi hin a blood essel a e
nega i e o E-cadhe in. IHC. Ba , 100 mm.
Please ci e his a icle as: Rod ı´guez-La go A e al., Epi helialeMesenchymal T ansi ion in a Case o Me as a ic Thy oid Ca cinoma in a B own Bea
(U sus a c os), Jou nal o Compa a i e Pa hology, h ps://doi.o g/10.1016/j.jcpa.2020.01.004
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YJCPA2305_p oo ■14 Feb ua y 2020 ■2/4
2A. Rod ı
´guez-La go e al.
like s uc u es and he umou was e en ually diag-
nosed as a me as a ic ollicula hy oid ca cinoma.
IHC using a a ie y o an ibodies demons a ed a
hy oid-de i ed epi helial umou . The p ima y and
pulmona y masses we e posi i e o mesenchymal
ma ke s and nega i e o he mos common hy oid
ma ke s (TTF-1 and TGB), indica ing a high le el
o anaplasia. Howe e , he me as a ic ad enal mass
e ained posi i i y o hese wo ma ke s and also o
E-cadhe in, indica ing a mo e di e en ia ed
neoplasm. Mul iple endoc ine neoplasia is cha ac e -
ized by he appea ance o di e en p ima y umou s
wi hin wo o mo e endoc ine glands a he same
ime (Roccabianca e al., 2006), a di e en scena io
o he p esen case.
Ec opic hy oid issue isno mally ound in a eas such
as he an e io ongue, achea and la ynx and such is-
sue migh appea in he ad enal gland (Shuno e al.,
2006). Rema kably, me as asis om hy oid ca ci-
nomas occasionally exhibi a be e -di e en ia ed
appea ance han he p ima y umou , some imes lead-
ing o misdiagnosis o ec opic hy oid issue a he han
neoplasia (Kondo e al., 2000; Shuno e al., 2006).
This case illus a es an example o umou cell plas-
ici y: a p ima y EMT in he hy oid gland ha
enabled he me as a ic p ocess, ollowed by a MET
ha allowed he neoplas ic cells o adap o a new
mic oen i onmen and o e-acqui e hei di e en i-
a ed epi helial cell ea u es in he ad enal gland.
Me as asis is a highly ine icien mul is ep p ocess,
only a small numbe o he ci cula ing neoplas ic cells
a e able o su i e and h i e in a o eign mic oen i-
onmen (Tsai and Yang, 2013). The ini ial me as a-
ic e en s o ca cinomas equen ly depend on EMT
ac i a ion, a pa ial ansi ion o he epi helial
neoplas ic cells in o a mesenchymal pheno ype ha
allows loss o cell- o-cell adhesion, disagg ega ion
and enhancemen o hei mo ili y and in asi e p op-
e ies (Kallu i and Weinbe g, 2009). In he p esen
case, EMT was demons a ed in he p ima y hy oid
mass and in he lung me as ases by he lowe ed
exp ession o E-cadhe in and he inc eased exp ession
o N-cadhe in and imen in, oge he wi h he acqui-
si ion o a spindle-shaped mesenchymal mo phology
by he neoplas ic cells (Knaack e al., 2018). In
con as , MET was demons a ed in he ad enal me-
as ases by he e-exp ession o E-cadhe in and he
loss o N-cadhe in and imen in signals, oge he
wi h he e-es ablishmen o an epi helial cell
mo phology. These ea u es allowed he ecogni ion
o he p ima y issue om which he umou a ose,
some hing ha was no easible in he me as ases in
he lungs. The in e ac ions be ween in ading
Fig. 2. Thy oid. Anaplas ic cells a e di usely posi i e o N-cad-
he in. IHC. Ba , 100 mm.
Fig. 3. Thy oid ca cinoma, me as asis in ad enal. Neoplas ic cells
a e di usely posi i e o E-cadhe in, s oma is nega i e.
IHC. Ba , 100 mm.
Fig. 4. Thy oid ca cinoma, me as asis in ad enal. Neoplas ic cells
a e nega i e o N-cadhe in. No mal ad enal co ex cells
exhibi s ong labelling. IHC. Ba , 250 mm.
Please ci e his a icle as: Rod ı´guez-La go A e al., Epi helialeMesenchymal T ansi ion in a Case o Me as a ic Thy oid Ca cinoma in a B own Bea
(U sus a c os), Jou nal o Compa a i e Pa hology, h ps://doi.o g/10.1016/j.jcpa.2020.01.004
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YJCPA2305_p oo ■14 Feb ua y 2020 ■3/4
Thy oid Ca cinoma in a B own Bea (U sus a c os)3
neoplas ic cells and he new mic oen i onmen can
modula e hei ou g ow h, p og ession and inal cell
mo phology and pheno ype (Gao e al., 2012). EMT
has been demons a ed in di e en neoplasms
including mamma y o panc ea ic ca cinomas
(Dong e and Weinbe g, 2019). The p esen case
shows a common adap i e p ocess in ca cinomas
ha possibly occu s in all species, and may explain
he umou cell plas ici y which enables coloniza ion
and g ow h o neoplasia in dis an o gan si es.
Acknowledgmen s
The cu en add ess o J. Asin is Cali o nia Animal
Heal h & Food Sa e y Labo a o y Sys em, Pa hol-
ogy, Mic obiology & Immunology Depa men , Uni-
e si y o Cali o nia a Da is, Da is, Cali o nia, USA.
The cu en add ess o M. Gimeno is Uni e si y Ve -
e ina y Teaching Hospi al Camden, The Uni e si y
o Sydney, Aus alia. The cu en add ess o P. Pinc-
zowski is NSW Depa men o P ima y Indus ies/
Biosecu i y & Food Sa e y, EMAI, Menangle,
Aus alia.
Q1
Supplemen a y da a
Supplemen a y da a o his a icle can be ound on-
line a h ps://doi.o g/10.1016/j.jcpa.2020.01.004.
Unci ed Re e ence
Q2
Rosol and Meu en, 2017.
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½
Recei ed, Sep embe 5 h, 2019
Accep ed, Janua y 20 h, 2020

Please ci e his a icle as: Rod ı´guez-La go A e al., Epi helialeMesenchymal T ansi ion in a Case o Me as a ic Thy oid Ca cinoma in a B own Bea
(U sus a c os), Jou nal o Compa a i e Pa hology, h ps://doi.o g/10.1016/j.jcpa.2020.01.004
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4A. Rod ı
´guez-La go e al.