Con en s lis s a ailable a ScienceDi ec
BBA - Molecula Basis o Disease
jou nal homepage: www.else ie .com/loca e/bbadis
Re iew
Mic oRNAs and ex acellula esicles in cholangiopa hies
☆
P. Olaizola
a,1
, P.Y. Lee-Law
a,b,c,1
, A. A belaiz
a
, A. Lapi z
a
, M.J. Pe ugo ia
a,b,d
, L. Bujanda
a,b,d
,
J.M. Banales
a,b,d,⁎
a
Depa men o Li e and Gas oin es inal Diseases, Biodonos ia Heal h Resea ch Ins i u e –Donos ia Uni e si y Hospi al, Uni e si y o he Basque Coun y (UPV/EHU),
San Sebas ian, Spain
b
Na ional Ins i u e o he S udy o Li e and Gas oin es inal Diseases (CIBERehd, “Ins i u o de Salud Ca los III”), Spain
c
Depa men o Gas oen e ology and Hepa ology, Radboud Uni e si y Medical Cen e, Nijmegen, Ne he lands
d
IKERBASQUE, Basque Founda ion o Science, Bilbao, Spain
ARTICLE INFO
Keywo ds:
mic oRNAs
Ex acellula esicles
Cholangiopa hies
Diagnosis
Pa hogenesis
The apy
ABSTRACT
Cholangiopa hies encompass a he e ogeneous g oup o diso de s affec ing bilia y epi helial cells (i.e. cho-
langiocy es). Ea ly diagnosis, p ognosis and ea men s ill emain clinically challenging o mos o hese dis-
eases and a e c i ical o adequa e pa ien ca e. In he pas decade, ex ensi e esea ch has emphasized
mic oRNAs (miRs) as po en ial non-in asi e bioma ke s and ools o accu a ely iden i y, p edic and ea
cholangiopa hies. MiRs can be eleased ex acellula ly conjuga ed wi h lipop o eins o encapsula ed in ex a-
cellula esicles (EVs). Resea ch on EVs is also gaining a en ion since hey a e p esen in mul iple biological
fluids and may ep esen a ele an sou ce o no el non-in asi e bioma ke s and be ehicles o new he apeu ic
app oaches. This e iew highligh s he mos p omising candida e miRs and EV- ela ed bioma ke s in cho-
langiopa hies, as well as hei ele an oles in bilia y pa hophysiology. This a icle is pa o a Special Issue
en i led: Cholangiocy es in Heal h and Disease edi ed by Jesus Banales, Ma co Ma zioni, Nicholas LaRusso and
Pe e Jansen.
Resea ch s a egy: PubMed sea ch (Ap il 2017) was done wi h he ollowing e ms: “mic oRNA”,“miRNA”,
“miR”,“ex acellula esicles”,“EV”,“exosomes”,“p ima y bilia y cholangi is”,“p ima y bilia y cholangi is”,
“PBC”,“p ima y scle osing cholangi is”,“PSC”,“cholangioca cinoma”,“CCA”,“bilia y a esia”,“BA”,“poly-
cys ic li e diseases”,“PLD”,“cholangiopa hies”,“choles a ic li e disease”. Mos significan a icles in ull- ex
English we e selec ed. The e e ence lis s o selec ed pape s we e also conside ed.
h p://dx.doi.o g/10.1016/j.bbadis.2017.06.026
Recei ed 16 May 2017; Recei ed in e ised o m 27 June 2017; Accep ed 28 June 2017
☆
This a icle is pa o a Special Issue en i led: Cholangiocy es in Heal h and Disease edi ed by Jesus Banales, Ma co Ma zioni, Nicholas LaRusso and Pe e Jansen.
⁎
Co esponding au ho a : Depa men o Li e and Gas oin es inal Diseases, Biodonos ia Heal h Resea ch Ins i u e –Donos ia Uni e si y Hospi al, Paseo del D . Begi is ain s/n, E-
20014 San Sebas ian, Spain.
1
Bo h au ho s con ibu ed equally o his wo k.
E-mail add ess: [email p o ec ed] (J.M. Banales).
Abb e ia ions: 15-PGDH, 15-hyd oxyp os aglandin dehyd ogenase; ADPKD, au osomal dominan polycys ic kidney disease; ADPLD, au osomal dominan polycys ic li e disease; AE2/
SLC4A2, CI
−
/HCO
3
–
anion exchange 2; ALP, alkaline phospha ase; AMA, an i-mi ochond ial an ibody; ARPKD, au osomal ecessi e polycys ic kidney disease; A s2, a senic esis ance
p o ein 2; ASGPR1, asialoglycop o ein ecep o 1; AUC, a ea unde he cu e; BA, bilia y a esia; CA19-9, cance an igen 19-9; CCA, cholangioca cinoma; CCL2, C-C mo i chemokine
ligand 2; Cdc25A, cell di ision cycle 25A; CDH6, cadhe in-6; CDK6, cyclin-dependen kinase 6; CHEK2, checkpoin kinase 2; c-Myc, myc p o o-oncogene p o ein; CXCL1, chemokine (C-X-
C mo i ) ligand 1; eCCA, ex ahepa ic cholangioca cinoma; EGFR, epide mal g ow h ac o ecep o ; ELK1, ETS domain-con aining p o ein Elk-1; EMT, epi helial mesenchymal ansi ion;
ERK, ex acellula signal- egula ed kinases; EV, ex acellula esicle; FDA, ood and d ug adminis a ion; FIBG, fib inogen gamma chain; FOXA1, o khead box p o ein A1; FOXO1,
o khead box p o ein O1; FXR, a nesoid X ecep o ; GSK3β, glycogen syn hase kinase-3 be a; HCC, hepa ocellula ca cinoma; HDAC4, his one deace ylase 4; HSC, hepa ic s ella e cell;
iCCA, in ahepa ic cholangioca cinoma; IGF1R, insulin-like g ow h ac o -1 ecep o ; IL, in e leukin; IL1β, in e leukin-1 be a; InsP3R3, ype III inosi ol 1,4,5- iphospha e ecep o ;
ITGB4, in eg in be a-4; MAPK, mi ogen ac i a ed p o ein kinase; MAP3K8, mi ogen-ac i a ed p o ein kinase kinase kinase 8; MBD2, me hyl-CpG-binding domain p o ein 2; Mcl-1,
induced myeloid leukemia cell diffe en ia ion p o ein Mcl-1; MDR1, mul id ug esis an p o ein 1; miR, mic oRNA; MMP, ma ix me allop o einase; MSC, mesenchymal s em cell; mTOR,
mechanis ic a ge o apamycin; MVB, mul i esicula body; NCAM1, neu al cell adhesion molecule 1; NDRG2, N-myc downs eam- egula ed gene 2; NUAK1, (nua) amily kinase 1;
PBMC, pe iphe al blood mononuclea cells; PBC, p ima y bilia y cholangi is; pCCA, pe ihila cholangioca cinoma; PDC-E2, py u a e dehyd ogenase complex-E2; PDCD4, p og ammed
cell dea h p o ein 4; Pe 1, pe iod ci cadian p o ein homolog 1; PLD, polycys ic li e disease; PSC, p ima y scle osing cholangi is; PSMD10, p o easome 26S subuni non-ATPase 10; PTEN,
phospha ase and ensin homolog; PTPN14, y osine-p o ein phospha ase non- ecep o ype 14; RB, e inoblas oma p o ein; RECK, e e sion-inducing cys eine- ich p o ein wi h Kazal
mo i s; RhoC, as homolog amily membe C; ROC, ecei e ope a ing cha ac e is ic; Smad4, small mo he s agains decapen aplegic homolog 4; SULT2A1,sulpho ans e ase 2A1; TET1,
en-ele en ansloca ion 1; TGFβR2, ans o ming g ow h ac o be a ecep o 2; TIMP3, me allop o einase inhibi o 3; TNFα, umo nec osis ac o alpha; TRAIL, TNF- ela ed apop osis-
inducing ligand; UDCA, u sodeoxycholic acid; VEGF, ascula endo helial g ow h ac o ; XIAP, X-linked inhibi o o apop osis p o ein
BBA - Molecula Basis o Disease 1864 (2018) 1293–1307
A ailable online 13 July 2017
0925-4439/ © 2017 Else ie B.V. All igh s ese ed.
T
1. In oduc ion
Bile duc epi helial cells (i.e. cholangiocy es) a e impo an in heal h
and disease. They ep esen a small p opo ion (3–5%) o he o al li e
cell popula ion bu , none heless, play essen ial oles o no mal li e
unc ion including he alkaliza ion and fluidiza ion o he p ima y bile
p oduced by hepa ocy es. Bilia y diseases, also e med as cholangio-
pa hies, encompass a wide spec um o e iologies comp ising gene ic,
in ec ious, immune-media ed, d ug-induced, ascula , neoplas ic o
idiopa hic. Al hough hei pa hogenesis s ill emains obscu e, ch onic
inflamma ion and choles asis seem o be common e en s ha exace -
ba e he wound-healing esponse leading o he de elopmen o li e
fib osis and ci hosis. Mos cholangiopa hies lack alid diagnos ic and/
o p ognos ic bioma ke s, as well as adequa e a ge s o he apy, de-
manding he need o u he esea ch. In he las yea s, he disco e y o
mic oRNAs (miRs) has ep esen ed a e olu ion and a pa adigm shi
(Fig. 1), pos ula ing hem as p omising bioma ke s and a ge s/ ools o
he apy. These small (18–23 nucleo ides) endogenous non-coding RNAs
play significan oles in mos physiological and pa hological cellula
e en s including p oli e a ion, diffe en ia ion, mig a ion, senescence
and su i al, by egula ing pos - ansc ip ional gene exp ession [1,2].
Cholangiopa hies display abe an miR signa u es in cholangiocy es,
immune cells, li e issue and biological fluids among o he s, e iden-
cing hei po en ial alue in diagnosis, p ognosis and he apy [3–5].
MiRs can be eleased in o he ex acellula medium associa ed wi h
lipop o eins o encapsula ed in ex acellula esicles (EVs), hus pa i-
cipa ing in in e cellula communica ion (Fig. 2)[6]. EVs a e small lipid-
enclosed sphe es sec e ed by many cell ypes and ound in mul iple
biological fluids including bile, blood and u ine. The e a e diffe en
ypes o EVs acco ding o hei o igin, size, molecula composi ion and
biological unc ion. Rega ding hei o igin, EVs a e classified in o
exosomes, plasma memb ane-de i ed esicles and apop o ic bodies.
Besides miRs, hey can also con ain o he nucleic acids, lipids and
p o eins. To da e, he mos s udied EVs a e exosomes, which a e e-
leased ex acellula ly upon exocy ic usion o endosome-de i ed mul-
i esicula bodies (MVBs) wi h he plasma memb ane o he cells. Then,
he exosomal con en can be u he deli e ed in o ecipien cells whe e
i can egula e gene exp ession and cellula unc ions. Changes in he
ansc ip omic and p o eomic EV con en ha e been epo ed in di -
e en cholangiopa hies, poin ing ou hei po en ial alue as non-in-
asi e bioma ke s (Fig. 1)[7].
This e iew p o ides cu en knowledge on he ole o miRs and EVs
in he pa hogenesis o bilia y diseases, and hei po en ial he apeu ic
alue. Mo eo e , he mos p omising miRs and EV- ela ed bioma ke s
as new non-in asi e diagnos ic and p ognos ic ools, and hei he -
apeu ic alue will be discussed. Finally, u u e di ec ions on basic and
clinical in es iga ions will be highligh ed.
2. Mic oRNAs in cholangiopa hies
2.1. Fib o-inflamma o y cholangiopa hies
P ima y bilia y cholangi is (PBC), p ima y scle osing cholangi is
(PSC) and bilia y a esia (BA) a e cholangiopa hies cha ac e ized by
ch onic inflamma ion, choles asis and bilia y fib osis. Along he disease
p og ession, ci hosis, po al hype ension and, ul ima ely, li e ailu e
may a ise in hese h ee diso de s.
2.1.1. P ima y bilia y cholangi is (PBC)
PBC is a ch onic choles a ic li e disease o unknown e iology linked
o au oimmune p ocesses a ge ing small and medium in ahepa ic bile
duc s. Wi hou ea men , PBC may p og ess o li e fib osis, ci hosis
and, ul ima ely, li e ailu e [8]. PBC mainly affec s middle-aged
women (~90%) and has been associa ed wi h en i onmen al oxins,
in ec ious agen s and ce ain gene ic ac o s [9]. Diagnosis is based on
clinical and se ological pa ame e s such as ele a ed le els o alkaline
phospha ase (ALP) and he p esence o specific an i-mi ochond ial an-
ibodies (AMAs) agains py u a e dehyd ogenase complex-E2 (PDC-E2)
in up o 95% o pa ien s [10,11]. Mo eo e , a high p opo ion o PBC
pa ien s also p esen s se um an i-nuclea an ibodies (ANAs) [12].
Howe e , indi iduals wi hou hese se ological ea u es equi e a li e
biopsy o de e mine he diagnosis. The chole e ic bile acid
Fig. 1. T ends o miR- and EV- ela ed a icles since 2000. A) Exponen ial g ow h in he numbe o miR- ela ed a icles (uppe le panel) and o al numbe o a icles ela ed o each ype o
cholangiopa hy (uppe igh panel). B) Exponen ial g ow h in he numbe o EV- ela ed a icles (lowe le panel) and o al numbe o a icles ela ed o each cholangiopa hy (lowe igh
panel). Red lines indica es he ends.
P. Olaizola e al. BBA - Molecula Basis o Disease 1864 (2018) 1293–1307
1294
u sodeoxycholic acid (UDCA) is he mains ay ea men , which is ad-
minis e ed daily and ch onically. None heless, depending on age, up o
50% o PBC pa ien s lack an adequa e esponse o UDCA ea men and
ha e lowe long- e m su i al han he gene al popula ion [13,14]. Fo
hose UDCA non- esponde s, he a nesoid X ecep o (FXR) agonis
obe icholic acid (OCA) has ecen ly been app o ed by he ood and
d ug adminis a ion (FDA) as mono he apy o combina ion wi h UDCA.
OCA imp o es some ma ke s o choles asis bu also induces side effec s
like p u i us [15]. The e o e, i is undamen al o iden i y accu a e
bioma ke s o specific and ea ly diagnosis, p ognosis and esponse o
he apy, as well as new he apeu ic a ge s o indi idualized pa ien
ca e in PBC.
Se e al s udies ha e in es iga ed he exp ession o miRs in se um
[16,17] and li e issue [18], as well as in specific cell ypes including
pe iphe al blood mononuclea cells (PBMCs) [19,20] and bilia y epi-
helial cells [21] om PBC pa ien s. The se um om PBC pa ien s was
cha ac e ized by an al e ed miR p ofile [17,22], wi h up egula ion o
bo h miR-122-5p and miR-141-3p, and down egula ion o miR-26b-5p
ha con e ed a highe diagnos ic alue han he se um le els o ALP
and ANAs (Table 1)[16]. In addi ion, deep sequencing analysis o
se um samples om PBC pa ien s iden ified miR-139-5p o be down-
egula ed in pa ien s wi h ad anced PBC s heal hy con ols and ended
o be lowe in ad anced PBC pa ien s s ea ly PBC pa ien s [23], sug-
ges ing i s po en ial alue o p edic he disease p og ession. The ole o
miRs as bioma ke s o p edic he esponse o ea men in PBC was
highligh ed by he ac ha o e exp ession o miR-299-5p in se um was
associa ed wi h non- esponse o UDCA ea men compa ed o bo h PBC
esponde s and heal hy con ols [20]. On he o he hand, al e a ions in
he miR exp ession p ofile we e also epo ed in PBMCs om PBC pa-
ien s [22]. In pa icula , down egula ion o miR-181a, miR-181b, miR-
374b, and miR-425 was ound in CD4
+
T cells om PBC pa ien s
compa ed o heal hy con ols. O no e, he down egula ion o miR-425
in CD4
+
T cells induced inflamma o y cy okines p oduc ion [24].
Choles asis is one o he main hallma ks o PBC and is caused,
pa ially, by down egula ion o bo h he CI
−
/HCO
3
–
anion exchange 2
(AE2/SLC4A2) and he ype III inosi ol 1,4,5- iphospha e ecep o
(InsP3R3) in cholangiocy es [25–29]. Rema kably, he e iopa hogenic
ole o AE2 in PBC was also highligh ed by he ac ha Ae2
−/−
mice
spon aneously de eloped diffe en PBC-like ea u es, including se um
specific AMAs [30,31]. One o he igge ing causes o bo h AE2 and
InsP3R3 down egula ion in PBC cholangiocy es is miR-506, which was
ound o e exp essed in hese cells and di ec ly a ge ed bo h mRNAs
leading o choles asis [21,32].Diffe en p o-inflamma o y cy okines
ound o e exp essed in PBC li e s such as in e leukins (ILs) 8, 12, 17,
18 and umo nec osis ac o alpha (TNFα) enhanced he ansc ip-
ional ac i i y o miR-506 gene p omo e in cholangiocy es, subse-
quen ly leading o al e ed exp ession o p o eins in ol ed in se e al
biological p ocesses, pa icula ly in mi ochond ial ene gy me abolism
[33]. MiR-506 induced PBC-like ea u es in cholangiocy es including
cell dediffe en ia ion, s ess, suscep ibili y o bile-sal induced apop-
osis, dys egula ion o mi ochond ial me abolism and PDC-E2 o e -
exp ession, ul ima ely p omo ing he ac i a ion and p oli e a ion o
PBMCs om PBC pa ien s when co-cul u ed [33]. These da a poin ou
he ele an ole o miR-506 in he e iopa hogenesis o PBC and i s
po en ial he apeu ic egula o y alue.
2.1.2. P ima y scle osing cholangi is (PSC)
PSC is a ch onic choles a ic li e disease affec ing bo h in a- and
ex ahepa ic la ge bile duc s [34,35]. Mos pa ien s a e middle-aged
men (~60%) and up o 80% also p esen inflamma o y bowel disease,
mos commonly ulce a i e coli is [8,34,35]. Impo an ly, PSC pa ien s
ha e a 400- old inc eased isk o de eloping cholangioca cinoma
(CCA) compa ed o he gene al popula ion [36]. Simila o PBC, he -
edi a y and en i onmen al elemen s a e associa ed wi h PSC, bu he
e iopa hogenesis emains unclea . The e is no effec i e medical ea -
men o al e he disease cou se and li e ansplan a ion is he only
cu a i e op ion [34,35,37]. In PSC, s udies on he pa hogenesis and ole
o miRs as bioma ke s o ools o he apy a e s ill limi ed.
A epo om 2016 poin ed ou he ele ance o se um miRs as
po en ial bioma ke s o he diagnosis o PSC and CCA (Table 1)[38].
Fig. 2. Biogenesis and elease o miRs. P ima y miRs (P i-miRs) a e syn hesized by RNA pol II o III (1). This ini ial RNA o m, wi h 5′cap and poly-A ail, is hen clea ed by he
mic op ocesso complex o med by DROSHA/DGCR8 gene a ing he hai pin-shaped P e-miR (P e-miRs) (2). The P e-miRs a e hen expo ed o he cy oplasm ia Expo in-5 (3), whe e
hey a e p ocessed by DICER/TRBP in o a double-s anded ma u e miR (4). One s and o he miR duplex can en e he RISC assembling pa hway (5) o be eleased by he cell, while he
complemen a y s and is o en deg aded. In o de o be eleased in o he ex acellula space, he ma u e miR s and can associa e o AGO 1–4 p o eins (6) o o lipop o eins (HDL, LDL)
(7). On he o he hand, miRs can be loaded in o exosomes ha a e ex acellula ly eleased upon exocy ic usion o MVB wi h he plasma memb ane (8) o in o mic o esicles o med by he
blebbing o he cellula plasma memb ane (9). Addi ionally, miRs can be discha ged in apop o ic bodies along wi h o he cellula -de i ed ma e ial (10).
RNA pol II: RNA polyme ase II; P e-miR: p ecu so miRNA; RISC: RNA-induced silencing complex; AGO: A gonau e p o ein; HDL: high densi y lipop o ein; LDL: low densi y lipop o ein;
MVB: mul i esicula body.
P. Olaizola e al. BBA - Molecula Basis o Disease 1864 (2018) 1293–1307
1295
In a disco e y phase, 21 miRs we e ound diffe en ially exp essed in
PSC, 33 in CCA and 26 in bo h g oups compa ed o heal hy con ols, as
well as 24 miRs in PSC s CCA wi h a ea unde he ecei e ope a ing
cha ac e is ic (ROC) cu e (AUC) > 0.700 [38]. A e a alida ion
phase in a second coho o pa ien s, miR-200c was confi med o be
down egula ed in PSC s heal hy con ols, whe eas inc eased le els o
bo h miR-483-5p and miR-194 we e ound in CCA s heal hy con ols as
well as bo h miR-483-5p and miR-222 in CCA s PSC [38]. Combina ion
o hese pa icula miRs u he imp o ed he specifici y and accu acy
o diagnosis.
In e ms o li e pa hophysiology, and pa icula ly in choles asis,
p egnane X ecep o (PXR) induced he exp ession o sulpho ans e ase
2A1 (SULT2A1) o con e li hocholic acid in o a less oxic o m and,
hus, p e en ed li e inju y. Howe e , PSC pa ien s a e cha ac e ized
by disease-specific impai men o SULT2A1 exp ession ollowing PXR
ac i a ion. In PSC, SULT2A1 exp ession migh be egula ed by miR-
378a-5p, which was ound o e exp essed in PSC s PBC li e s and was
p edic ed by bioin o ma ics ools o a ge SULT2A1 gene exp ession
[39]. On he o he hand, i was epo ed ha miR-21 p omo es bilia y
hype plasia [40] and miR-7a enhances cholangiocy e p oli e a ion [41]
in animal models o choles asis and scle osing choles asis, espec i ely,
sugges ing a po en ial ole o hese miRs in he pa hogenesis o PSC.
Howe e , he ole o hese o o he miRs in he PSC e iopa hogenesis
emains s ill unknown.
2.1.3. Bilia y a esia (BA)
BA is a p og essi e and des uc i e cholangiopa hy affec ing he
in a- and ex ahepa ic bile duc s o neona es and ha causes se e e
choles asis [42]. The e iopa hogenesis o his disease emains s ill ob-
scu e bu se e al gene ic and en i onmen al ac o s ha e been pos u-
la ed o pa icipa e in i s de elopmen [43]. Wi hou ea men , BA
pa ien s seldom su i e mo e han 2 yea s [42]. To da e, he only e -
ec i e he apy is he Kasai po oen e os omy, which es o es he bile
flow. Howe e , ea ly diagnosis and in e en ion a e c ucial o he
p ognosis i his disease. BA is diagnosed by ope a i e cholangiog aphy
and/o li e biopsy, which a e in asi e and ime consuming, high-
ligh ing he u gen need o non-in asi e al e na i es.
Se um o BA pa ien s may con ain p omising miR bioma ke s wi h
high sensi i i y and specifici y o he diagnosis (Table 1). Indeed, up-
egula ion o miR-200a, miR-200b and miR-429 (miR-200 clus e ) was
epo ed in BA pa ien s compa ed o o he o ms o neona al hype bi-
li ubinemia (NH) [44]. On he o he hand, miR-4429 down egula ion
and miR-4689 up egula ion we e also epo ed in ano he s udy wi h
po en ial diagnos ic alue [45]. Addi ionally, by using nex -gene a ion
sequencing, miR-140-3p was ound down egula ed in plasma o BA
pa ien s compa ed o choles a ic disease pa ien s and heal hy con ols,
showing diagnos ic po en ial [46]. In e es ingly, he same s udy ied o
iden i y p e iously epo ed dys egula ed miRs (i.e.miR-200 amily
[44], miR-21 [47,48], miR-29a [48,49], miR-222 [47,50,51]) im-
plica ed in human and expe imen al BA, howe e only up egula ion o
miR-200 amily was alida ed [46].
Diffe en dys egula ed miRs in BA a e belie ed o be associa ed wi h
p o-inflamma o y and p o-fib o ic p ocesses (Table 2)[22]. MiR-19b,
which was ound down egula ed in li e issue o BA pa ien s [52],
Table 1
MiRs as diagnos ic bioma ke s o cholangiopa hies.
AUC, a ea unde he cu e; BA, bilia y a esia; CA19-9, cance an igen 19-9; CCA, cholangioca cinoma; iCCA, in ahepa ic cholangioca cinoma; MiR, mic oRNA; PBC, p ima y bilia y
cholangi is; PSC, p ima y scle osing cholangi is; SEN, sensi i i y; SPE, specifici y.
Disease MiR Exp ession Sou ce Numbe o pa ien s SEN (%) SPE (%) AUC Re e ence
PBC 122-5p + 141-3p
+ 26b-5p
Up (122-5p +141-3p)
Down (26-5p)
Se um PBC (n = 82) s heal hy con ols (n = 60) 80.5 88.3 0.905 [16]
PSC 200c Down Se um PSC (n = 40) s heal hy con ols (n = 40) ––0.740 [38]
BA 200a Up Se um BA (n = 24) s o he o ms o neona al hype bili ubinemia (n = 24) 83.3 83.3 0.862 [44]
200b Up Se um 79.2 79.2 0.807
429 Up Se um 70.8 91.7 0.806
140-3p Down Plasma BA (n = 44) s choles a ic disease con ols (n = 20) and heal hy con ols
(n = 20)
66.7 79.1 0.750 [46]
4429 Down Se um BA (n = 35) s non-BA neona al choles asis (n = 20) 83.3 80.0 0.789 [45]
4689 Up Se um 66.7 80.0 0.722
CCA 9 Up Bile CCA (n = 7) and gallbladde cance (n = 2) s choledocholi hiasis
pa ien s (n = 9)
88.9 100.0 0.975 [67]
145 Up Bile 77.8 100.0 0.975
942 Up Bile 77.8 100.0 0.765
302c Up Bile 88.9 100.0 e
199a-3p Up Bile 88.9 100.0 e
222 Up Bile 88.9 100.0 e
105 Up Bile 77.8 100.0 e
21 Up Se um iCCA (n = 74) s heal hy con ols (n = 74) 87.8 90.5 0.908 [68]
21 Up Plasma iCCA (n = 25) s heal hy con ols (n = 7) ––0.940 [71]
26a
a
Up Se um CCA (n = 66) s heal hy con ols (n = 66) 84.8 81.8 0.899 [73]
106a
a
Down Se um CCA (n = 103) s heal hy con ol (n = 20) 81.6 85.0 0.890 [76]
150 Up Plasma iCCA (n = 15) s heal hy con ols (n = 15) 80.6 58.1 0.791 [75]
192 Up Se um O. i e ini CCA (n = 10) s heal hy con ols (n = 32) 74.0 72.0 0.809 [79]
21 + 192 Up U ine O. i e ini CCA (n = 22) s heal hy con ols (n = 21) 81.8 71.4 0.849 [80]
483-5p + 194 Up Se um CCA (n = 40) s heal hy con ols (n = 40) ––0.810 [38]
483-5p + 222 Up Se um CCA (n = 40) s PSC (n = 40) ––0.770
1281 Down Se um CCA (n = 31) s PSC (n = 40) 55.0 90.0 0.830 [81]
126 Down Se um 68.0 93.0 0.870
26a Down Se um 52.0 93.0 0.780
30b Down Se um 52.0 88.0 0.780
122 Down Se um 32.0 90.0 0.650
412 Up Bile PSC/CCA (n = 12) s PSC (n = 52) 50.0 89.0 0.810 [81]
640 Up Bile 50.0 92.0 0.810
3189 Up Bile 67.0 89.0 0.800
1537 Up Bile 67.0 90.0 0.780
1537 + CA19-9 Up Bile 73.0 93.0 0.910
a
Also p ognos ic bioma ke .
P. Olaizola e al. BBA - Molecula Basis o Disease 1864 (2018) 1293–1307
1296
di ec ly a ge ed ans o ming g ow h ac o be a ecep o 2 (TGFβR2)
gene exp ession and is belie ed o indi ec ly induce downs eam TGFβ
signaling, which is in ol ed in hepa ic s ella e cells (HSCs) ac i a ion
[52]. Addi ionally, inc eased miR-222, miR-200b and miR-21 in li e
issue o BA pa ien s may also s imula e HSCs ac i a ion p omo ing fi-
b osis [47,50,53]. On he o he hand, he p o-inflamma o y cy okine
IL6, up egula ed in li e o BA pa ien s, has been epo ed o enhance
cholangiocy e p oli e a ion h ough miR-124 and miR-200 amily [54].
2.2. Polycys ic li e diseases (PLD)
PLD comp ise a he e ogeneous g oup o congeni al cholangiopa hies
inhe i ed in dominan [i.e. au osomal dominan polycys ic li e disease
(ADPLD) o au osomal dominan polycys ic kidney disease (ADPKD)] o
ecessi e o m [i.e. au osomal ecessi e polycys ic kidney disease
(ARPKD)] and cha ac e ized by p og essi e de elopmen o mul iple
fluid-filled bilia y cys s (> 10), which a e he main cause o mo bidi y
[55–57]. PLD can be ound isola ed (e.g. ADPLD) o associa ed wi h
enal cys ogenesis (e.g. ADPKD and ARPKD). Cu en su gical and/o
pha macological ea men s do no imp o e he p ognosis o hese
diseases, and li e ansplan a ion emains he only cu a i e op ion.
Hepa ic cys ogenesis in PLD is cha ac e ized by unc ional al e a ions in
bile duc epi helial cells [58] ha include miRs dys egula ion [59,60].
To da e, he e is only a p e ious epo highligh ing he impo an ole
o miRs in PLD pa hophysiology [59,60]. In his s udy, an abno mal
miR exp ession p ofile was ound in cholangiocy es isola ed om an
animal model o ARPKD (i.e. he PCK a ), which has he same human
o hologous gene mu a ed (i.e.PKHD1). O no e, mos o he dys egu-
la ed miRs we e ound down egula ed in PCK cholangiocy es [59].O
pa icula in e es is miR-15a, which was ound highly down egula ed
in bo h a and human cys ic cholangiocy es [59]. MiR-15a di ec ly
a ge s cell di ision cycle 25A (Cdc25a) p omo ing cys ic cholangiocy e
cell p oli e a ion [59]. In e es ingly, expe imen al a ge ing o miR-15a
wi h specific an i-sense oligonucleo ides inhibi ed cys ic cholangiocy es
g ow h [59].
2.3. Cholangioca cinoma (CCA)
CCA includes a he e ogeneous g oup o malignancies wi h bilia y
diffe en ia ion ea u es ha may a ise om diffe en li e cell ypes
including ma u e cholangiocy es. CCA is he second mos equen li e
umo accoun ing o 10–20% o all p ima y li e neoplasms [61,62].
A ending o hei ana omical loca ion, CCAs a e classified as in-
ahepa ic (iCCA), pe ihila (pCCA) and ex ahepa ic (eCCA) [63].
Epidemiological s udies indica e ha CCA wo ldwide incidence has
been ising in he las decades [63,64] anging om 0.30 o 8.75 pe
100.000 indi iduals depending on he geog aphical a ea [61,63,64].
Se e al isk ac o s, including PSC, ci hosis, i al hepa i is B and C,
hepa oli hiasis, congeni al bilia y mal o ma ions as well as he hepa-
obilia y flukes endemic in Eas Asia (i.e.Opis ho chis i e ini and
Clonochis sinensis), ha e been iden ified o CCA [64]. Howe e , he
e iology o mos CCAs s ill emains unknown [63,64]. Since CCAs a e
o en asymp oma ic in ea ly s ages, diagnosis is usually conduc ed
when he disease is ad anced and widesp ead [63,64]. The cu en
diagnos ic s a egy comp ises a combina ion o imaging me hods, non-
specific umo bioma ke s in se um [i.e. ca bohyd a e an igen 19-9
(CA19-9)] and his ological analyses o umo biopsies. La e diagnosis
comp omises he po en ial cu a i e op ions, which a e mainly based on
su ge y, leading o poo p ognosis [63,64]. Fu he mo e, he espon-
si eness o CCA o cu en chemo he apies is e y limi ed [63,64].
The e o e, ea ly de ec ion o hese umo s is c ucial o hose pa ien s
wi h isk ac o s and o hose ha p esen ecu ence a e su ge y.
Du ing he las decade, a significan numbe o esea ch a icles ha e
been published on he ole o miRs in CCA (Fig. 1A) poin ing ou hei
ele an alue as non-in asi e bioma ke s (Table 1) and po en ial a -
ge s o he apy.
2.3.1. Mic oRNAs as bioma ke s
To s eng hen he diagnosis o CCA and moni o umo p og ession,
ele a ed CA19-9 le els in se um a e commonly used as a com-
plemen a y app oach o imaging me hods. Howe e , he sensi i i y and
specifici y o his non-in asi e bioma ke is modes , pa icula ly in
ea ly s ages o he disease [65,66].
Inc easing e idence poin s ou he ele ance o miRs as bioma ke s
o CCA. Indeed, se e al dys egula ed miRs ha e been desc ibed in
se um, plasma, u ine o bile om CCA pa ien s, showing high AUC
alues o diagnosis (Table 1). High- h oughpu eal- ime PCR-based
assays pe o med in human bile samples showed ha miR-9, miR-105,
miR-145, miR-199-3p, miR-222, miR-302c and miR-942 le els we e
highe in bo h CCA and gallbladde cance compa ed o pa ien s wi h
choledocholi hiasis [67] and acknowledged miR-9 as a eliable diag-
nos ic indica o o bilia y ac cance [67]. In addi ion, diffe en
s udies e ealed miR-21 as a po en ial bioma ke candida e o he di-
agnosis o iCCA [68,69],as i was ound o e exp essed in bo h se um
and plasma om hese pa ien s compa ed o heal hy indi iduals
[70,71]. Clinical s age and umo diffe en ia ion deg ee we e epo ed
o co ela e wi h he le els o miR-21 in CCA issue, and high miR-21
exp ession has been linked o poo o e all su i al, e idencing i s
p omising ole as a p ognos ic bioma ke [70,72]. Simila ly, le els o
miR-26a we e shown o be inc eased in CCA issues, cell lines and
Table 2
MiRs in ol ed in non- umo cholangiopa hies.
AE2, CI
−
/HCO
3
–
anion exchange 2; BA, bilia y a esia; Cdc25a, cell di ision cycle 25A; FOG2, iend o Ga a 2; FOXA2, o khead box p o ein A2; IGF1: insulin-like g ow h ac o 1;
IGF1R, insulin-like g ow h ac o 1 ecep o ; IL6R, in e leukin 6 ecep o ; InsP3R3, ype III inosi ol 1,4,5- iphospha e ecep o ; Ngn-3, neu ogenin-3; N-Ras, neu oblas oma RAS i al
oncogene homolog; PBC, p ima y bilia y cholangi is; PLD, polycys ic li e disease; PPP2R2A, p o ein phospha ase 2 egula o y subuni B alpha; PSC, p ima y scle osing cholangi is;
PTEN, phospha ase and ensin homolog; Smad, small mo he s agains decapen aplegic; STAT3, signal ansduce and ac i a o o ansc ip ion 3; TGFβ, ans o ming g ow h ac o be a;
TNFα, umo nec osis ac o alpha.
Disease MiR Exp ession Ta ge Func ion Sample Re e ence
PBC 506 Up AE2, InsP3R3 Sec e ion Cell lines, issue [21,33]
139-5p Down c-FOS, TNF-αInflamma ion Tissue, se um [23]
425 Down N-Ras Inflamma ion Se um [24]
PSC 7a Up Ngn-3 P oli e a ion Animal model [41]
21 Up Smad Fib osis, p oli e a ion Tissue, animal model [40]
BA 21 Up PTEN Fib osis Tissue [47]
29a Up IGF-1, IGF-1R Cell dea h, inflamma ion Animal model [49]
200a, 200b, 200c Up FOXA2 Inflamma ion, p oli e a ion Tissue, animal model [44]
200b Up FOG2 P oli e a ion, mig a ion, fib osis Tissue [53]
222 Up PPP2R2A Fib osis Tissue, animal model [50,51]
19b Down TGFβFib osis Tissue [52]
124 Down STAT3, IL-6R Inflamma ion, p oli e a ion Tissue, animal model [54]
PLD 15a Down Cdc25a P oli e a ion Cell lines, issue [59]
P. Olaizola e al. BBA - Molecula Basis o Disease 1864 (2018) 1293–1307
1297
se um om CCA pa ien s compa ed o heal hy con ols [73,74]. Fu -
he mo e, miR-150 le els we e ound up egula ed in plasma om iCCA
pa ien s [75] and i s combina ion wi h CA19-9 imp o ed he indi idual
diagnos ic capaci y o bo h bioma ke s [75]. In con as , he exp ession
o se um ci cula ing miR-106a was epo ed down egula ed in CCA
compa ed o heal hy con ols, bu i s diagnos ic alue was lowe han
CA19-9 [76]. As p e iously men ioned o PSC-CCA, se um miR-483-5p
and miR-194 we e ound up egula ed in CCA pa ien s compa ed o
con ol indi iduals and hei combina ion imp o ed he diagnos ic
po en ial o each miR [38]. Rema kably, inc eased se um le els o miR-
222 and miR-483-5p we e iden ified in CCA s PSC pa ien s [38].A
diffe en s udy p o ided a panel o 5 se um miRs (miR-1281, miR-126,
miR-26a, miR-30b and miR-122) able o disc imina e be ween PSC and
CCA pa ien s [77]. Fu he mo e, a dis inc miR exp ession pa e n was
iden ified in bile om PSC-CCA s PSC pa ien s (miR-412, miR-640,
miR-1537 and miR-3189) [77]. In e es ingly, combina ion o miR-1537
and CA19-9 le els displayed highe diagnos ic capaci y han CA19-9
alone [77].
MiRs ha e also been pos ula ed as p ognos ic bioma ke s o CCA.
Down egula ion o miR-106a in se um om CCA pa ien s was asso-
cia ed wi h highe isk o lymph node me as asis and in e sely co e-
la ed wi h o e all su i al [76]. Likewise, down egula ion o miR-150-
5p in plasma, bile and umo issue om CCA pa ien s nega i ely co -
ela ed wi h CA19-9 le els and umo g ade [78]. On he o he hand,
inc eased se um le els o miR-192 we e associa ed wi h me as asis and
poo o e all su i al in li e fluke-associa ed CCA pa ien s compa ed o
heal hy con ols [79]. O no e, inc eased miR-192 and miR-21 le els
we e ound in u ine om li e fluke-associa ed CCA pa ien s and hei
combina ion imp o ed he diagnos ic powe o hese wo miRs alone
[80].
The o e all diagnos ic capaci y o miRs in CCA was emphasized in
wo independen me a-analysis, whe e he pooled o 11 miRs om 8
independen s udies displayed AUC alues o 0.900 and 0.880, e-
spec i ely [81,82]. miRs in bile and se um showed highe diagnos ic
alue (i.e. 0.957 and 0.957, espec i ely) han issue (0.847) and u ine
(0.745) [81].
2.3.2. Mic oRNAs in CCA pa hology
Abe an ly exp essed miRs in CCA umo cells ha e been desc ibed
o pa icipa e in pa hological p ocesses including cell p oli e a ion,
diffe en ia ion, su i al, in asion/mig a ion, epi helial mesenchymal
ansi ion (EMT), epigene ics and chemo esis ance (Fig. 3)[22]. These
miRs can unc ion as oncogenes o umo supp esso s (Table 3).
2.3.3. Onco-mic oRNAs
Se e al up egula ed miRs in CCA cells unc ion as onco-miRs p o-
mo ing umo g ow h. Fo ins ance, miR-21 s ands ou as a pi o al
egula o o se e al pa hophysiological p ocesses such as cell p o-
li e a ion, su i al, EMT, in asi eness/me as asis and chemo esis ance
ia di ec a ge ing o diffe en umo supp esso s including p o-
g ammed cell dea h p o ein 4 (PDCD4), me allop o ease inhibi o 3
(TIMP3), e e sion-inducing cys eine- ich p o ein wi h Kazal mo i s
(RECK), y osine-p o ein phospha ase non- ecep o ype 14 (PTPN14),
15-hyd oxyp os aglandin dehyd ogenase (15-PGDH), phospha ase and
ensin homolog (PTEN) and phospha idylinosi ol 3-kinase (PI3K)
[68,70,83–86], among o he s [87]. In addi ion, a senic esis ance p o-
ein 2 (A s2), in ol ed in miR biogenesis, was ound o e exp essed in
CCA issue and cell lines leading o inc eased miR-21 exp ession, which
in u n inhibi ed i s downs eam a ge s u he con ibu ing o CCA cell
p oli e a ion and oncogenesis [69].
The mRNA o diffe en genes in ol ed in he blockade o cell cycle
such as p15, p21 and Cyclin E1 has been epo ed o be di ec ly a ge ed
by he oncogenic miR-224, leading o enhanced cell cycle p og ession
and umo g ow h [88]. Ano he miR wi h a po en ial ole in cell cycle
p og ession is miR-34a, which was ound o di ec ly inhibi pe iod
ci cadian p o ein homolog 1 (Pe 1) exp ession and i s in e ac ion wi h
checkpoin kinase 2 (CHEK2), p e en ing cell cycle a es [89]. Eme -
ging da a ha e demons a ed ha miR-191 di ec ly a ge s en-ele en
ansloca ion 1 (TET1), which in u n deme hyla es and he eby e-
p esses p53 exp ession in CCA cells. Hence, upon miR-191 up egula-
ion, he umo supp esso ac i i y o p53 is inhibi ed and leads o cell
su i al and p oli e a ion [90]. Fu he mo e, miR-429 exp ession was
epo ed o be inc eased in CCA, co ela ing wi h he hypome hyla ed
s a us o i s p omo e [91]. Func ional in i o s udies showed ha miR-
429 di ec ly a ge ed he umo supp esso cadhe in-6 (CDH6) inducing
umo cell g ow h [91]. Likewise, inc eased cellula p oli e a ion and
apop osis e asion we e shown in iCCA upon miR-31 up egula ion
h ough RAS p21 GTPase ac i a ing p o ein 1 (RASA1) di ec inhibi ion
[92]. RASA1 supp ession led o inc eased le els o he ac i e RAS o m
(GTP-bound) as well as o inc eased ERK1/2 phospho yla ion u he
ac i a ing he MAPK signaling pa hway and ul ima ely enhancing
umo g ow h [92]. MiR-421 was iden ified as ano he onco-miR in
CCA ha p omo ed umo cell p oli e a ion, mig a ion and colony
o ming ia FXR inhibi ion [93]. CCA cell su i al was associa ed wi h
miR-25, which p o ec ed umo cells agains TNF- ela ed apop osis-in-
ducing ligand (TRAIL)-media ed apop osis ia di ec a ge ing o TRAIL
Dea h Recep o -4 (DR4) [94].
The exp ession o he umo supp esso N-myc downs eam- egu-
la ed gene 2 (NDRG2) was ound o be ep essed by miR-181c, esul ing
in cell cycle p og ession, cell p oli e a ion and chemo esis ance [95].
Addi ionally, miR-181c co ela ed wi h he exp ession o mesenchymal
ma ke s N-cadhe in and imen in, and nega i ely co ela ed wi h he
epi helial ma ke E-cadhe in, he e o e, p omo ing EMT [95]. Mo e-
o e , miR-221 was shown o a ge he umo supp esso PTEN and
p omo e mig a ion and in asion in CCA cells h ough he β-ca enin
signaling pa hway-media ed EMT, as i a o ed β-ca enin ansloca ion
in o he nucleus. Mo eo e , β-ca enin could ac i a e c-Jun, known o
induce miR-221 exp ession. Thus, miR-221, β-ca enin and c-Jun sig-
naling pa hways o m a posi i e eedback loop h ough PTEN inhibi ion
ha enhances EMT [96]. Finally, miR-24 has been ound o ac as an
onco-miR by pa ially ep essing he umo supp esso p o ein menin
and, hence, s imula ing p oli e a ion, mig a ion, in asi eness and an-
giogenesis in CCA umo s [97].
2.3.4. Tumo supp esso mic oRNAs
Diffe en umo supp esso miRs a e down egula ed in CCA cells
leading o umo g ow h. In pa icula , down egula ion o miR-494 in
CCA cells induced he exp ession o i s di ec a ge cyclin-dependen
kinase 6 (CDK6) leading o cell cycle p og ession [98,99]. Simila ly,
miR-122 was able o inhibi he exp ession o genes in ol ed in cell
cycle p og ession including Cyclin G1 and insulin-like g ow h ac o 1
ecep o (IGF1R), and i s baseline down egula ion in CCA esul ed in
cell p oli e a ion [100]. Rega ding cell su i al, dec eased miR-29b
le els in CCA cells p omo ed he exp ession o i s a ge induced mye-
loid leukemia cell diffe en ia ion p o ein Mcl-1 (Mcl-1), an an i-apop-
o ic Bcl-2 amily membe ha p o ec s cells agains TRAIL-media ed
p og ammed cell dea h [101]. Simila ly, dec eased miR-410 exp ession
in CCA cells esul ed in he up egula ion o i s a ge , he X-linked in-
hibi o o apop osis p o ein (XIAP), leading o umo cell g ow h and
in asi eness [102]. Likewise, educed miR-212 le els in CCA cell lines
led o he up egula ion o i s di ec a ge o khead box p o ein A1
(FOXA1), p omo ing inc eased umo cell p oli e a ion and in asion
[103]. On he o he hand, miR-145 was poin ed ou as ano he umo
supp esso ound down egula ed in CCA cells [104]. MiR-145 p e en s
he umo g ow h, p oli e a ion and in asion o iCCA cells by di ec ly
a ge ing no el (nua) amily kinase 1 (NUAK1), which u he nega-
i ely egula ed Ak /Fo khead box p o ein O1 (FOXO1) pa hway and
ma ix me allop o einase (MMP) exp ession hal ing iCCA p og ession
[104]. Mo eo e , dec eased miR-150-5p le els in CCA cells p omo ed
cell p oli e a ion, mig a ion and in asion ia up egula ion o i s di ec
a ge he oncogenic ETS domain-con aining p o ein Elk-1 (ELK1) [78].
Rega ding EMT, mig a ion and in asi eness, dec eased miR-214
P. Olaizola e al. BBA - Molecula Basis o Disease 1864 (2018) 1293–1307
1298
le els we e epo ed in me as a ic iCCA issues compa ed o non-me-
as a ic ones, leading o dec eased exp ession o he epi helial ma ke E-
cadhe in and induced exp ession o he EMT ansc ip ion ac o Twis
[105]. In addi ion, down egula ion o miR-122 in human CCA has been
sugges ed o be in ol ed in umo cell mig a ion and in asion h ough
dys egula ion o MMP2, MMP9, RECK, E-cadhe in and N-cadhe in ex-
p ession [100]. The ac i a ion o he ERK/MMP2/MMP9 signaling
pa hway and he consequen p oli e a ion, mig a ion and in asion we e
induced upon miR-138 down egula ion in CCA cells ia Ras homolog
gene amily membe C (RhoC) up egula ion [106]. Simila ly, miR-200c
was epo ed o media e EMT in CCA cells ia di ec a ge ing o neu al
cell adhesion molecule 1 (NCAM1) [107]. On he o he hand, he epi-
de mal g ow h ac o (EGF)-dependen mig a ion o iCCA cells was
associa ed wi h down egula ed miR-376c exp ession ia up egula ion o
i s di ec a ge g ow h ac o ecep o -bound p o ein 2 (GRB2). Ad-
di ionally, in e leukin-1 be a (IL1β) and MMP9 we e sugges ed o be
unc ioning downs eam o GRB2 signaling [108]. Likewise, TGFβmay
be esponsible o he educed miR-29a le els in CCA cell lines and
issue, leading o he s imula ion o umo g ow h and me as asis ia
ac i a ion o i s di ec a ge his one deace ylase 4 (HDAC4) [109].
Inflamma ion has been es ablished as a ele an componen o
umo p og ession. MiR-605 was ound down egula ed in iCCA issue
leading o o e exp ession o i s di ec a ge 26S p o easome ATPase
egula o y subuni 10 (PSMD10), which p e en ed e inoblas oma
p o ein (RB) om inhibi ing in e leukin 6 (IL6) [110]. Induc ion o he
IL6/STAT3 pa hway ul ima ely p omo ed cell p oli e a ion and in a-
sion in i o and umo g ow h in i o h ough he up egula ion o Cy-
clin D1, ascula endo helial g ow h ac o (VEGF), MMP2 and MMP9
[110,111]. The IL6/STAT3 pa hway has also been desc ibed as he
a ge o he le -7/miR-99a/miR-125b clus e , which was ound
down egula ed in CCA issues [112]. Essen ially, his miR clus e di-
ec ly a ge ed cen al inflamma o y elemen s including IL6, IL6R and
IGF1R, which in u n ac i a ed STAT3 downs eam signaling enhancing
CCA p og ession [112]. Down egula ion o miRs in CCA may be caused
by hype me hyla ion o hei p omo e s. Dec eased miR-370 le els in
CCA cells we e linked o hype me hyla ion o i s p omo e ia IL6-de-
penden ac i a ion o DNA me hyl ans e ases. Consequen ly, he ex-
p ession o miR-370 was educed and he exp ession o i s a ge he
mi ogen-ac i a ed p o ein kinase kinase kinase 8 (MAP3K8) was en-
hanced con ibu ing o umo g ow h [113]. In addi ion, he pa e nal
allele o miR-370 was o en ound silenced h ough genomic imp in ing
in CCA pa ien s [114]. Mo eo e , due o he o e exp ession o IL6 in
Fig. 3. Abe an miR exp ession in he pa hogenesis o CCA.
Dys egula ed miRs in umo igenic cellula e en s: cell cycle
dys egula ion and p oli e a ion (A); EMT, mig a ion and
in asion (B); angiogenesis (C); cell dea h (D); inflamma ion
(E); chemo esis ance (F) and epigene ics (G). The cen al
image co esponds o a human li e wi h CCA. Up egula ed
miRs a e shown in blue colo and down egula ed ones in ed
colo .
CCA, cholangioca cinoma; M1, mac ophage ype 1; M2,
mac ophage ype 2; TRAIL, TNF- ela ed apop osis-inducing
ligand; VEGF, ascula endo helial g ow h ac o .
P. Olaizola e al. BBA - Molecula Basis o Disease 1864 (2018) 1293–1307
1299
Table 3
miRs in ol ed in bilia y umo igenesis.
15-PGDH, 15-hyd oxyp os aglandin dehyd ogenase; CCA, cholangioca cinoma; CDH6, cadhe in-6; CDK6, cyclin-dependen kinase 6; c-Me , y osine-p o ein kinase Me ; c-Myc, myc p o o-oncogene p o ein; DR4, dea h ecep o 4; ELK1, ETS
domain-con aining p o ein Elk-1; EMT, epi helial mesenchymal ansi ion; FOXA1, o khead box p o ein A1; FXR, a nesoid X ecep o ; GRB2, g ow h ac o ecep o -bound p o ein 2; GSK3B, glycogen syn hase kinase 3 be a; HDAC4, his one
deace ylase 4; iCCA, in ahepa ic cholangioca cinoma; IGF1R, insulin-like g ow h ac o 1 ecep o ; IL6, in e leukin 6; IL6R, in e leukin 6 ecep o ; KRT19, ke a in 19; MAP3K8, mi ogen-ac i a ed p o ein kinase kinase kinase 8; MBD2, me hyl-CpG-
binding domain p o ein 2; Mcl-1, induced myeloid leukemia cell diffe en ia ion p o ein Mcl-1; miR, mic oRNA; MMP2, ma ix me allop o einase 2; MMP9, ma ix me allop o einase 9; mTOR, mechanis ic a ge o apamycin; NCAM1, neu al cell
adhesion molecule 1; NDRG2, N-myc downs eam- egula ed gene 2; NUAK1, NUAK amily kinase 1; PDCD4, p og ammed cell dea h p o ein 4; Pe 1, pe iod ci cadian p o ein homolog 1; PIK3R, phosphoinsi ide-3-kinase egula o y subuni 1;
PSMD1, p o easome 26S non-ATPase egula o y subuni 1; PTEN, phospha ase and ensin homolog; PTPN14, y osine-p o ein phospha ase non- ecep o ype 14; PTTG1, pi ui a y umo - ans o ming gene 1 p o ein; RASA1, RAS p21 p o ein
ac i a o 1; RECK, e e sion-inducing cys eine- ich p o ein wi h Kazal mo i s; RhoC, as homolog amily membe C; Smad4, small mo he s agains decapen aplegic homolog 4; TET1, en-ele en ansloca ion 1; TIMP3, me allop o einase inhibi o 3;
TOP2A, DNA opoisome ase 2-alpha; Twis , wis basic helix-loop-helix ansc ip ion ac o ; WNT10B, P o ein Wn -10B; XIAP, X-linked inhibi o o apop osis p o ein.
ROLE IN
TUMORIGENESIS MiR DISEASE TARGET FUNCTION SAMPLE REFERENCE
Onco-miR
21 iCCA PTPN14, PTEN P oli e a ion, umo g ow h, apop osis [69]
21 O. i e ini iCCA PDCD4 P oli e a ion, umou g ow h, mig a ion, oncogenesis, model [72]
21 CCA PDCD4, TIMP3, E-Cadhe in, N-Cadhe in, Vimen in, PTEN,
PDCD4, RECK, 15-PGDH chemo esis ance Cell lines, issue [68, 83-86]
24 CCA Menin P oli e a ion, mig a ion, in asion, angiogenesis [97]
25 CCA DR4 Apop osis Cell lines, issue [94]
26a CCA GSK-3b P oli e a ion Cell lines, issue [74]
31 iCCA RASA1 P oli e a ion, apop osis Cell lines, issue [92]
34a CCA Pe 1 P oli e a ion, cell cycle p og ession, umo g ow h, in asion Cell lines, issue [89]
181c CCA NDRG2 P oli e a ion, umo g ow h, EMT, mig a ion, in asion,
chemo esis ance, senescence Cell lines, issue [95]
191 iCCA TET1 P oli e a ion, cell cycle p og ession, umo g ow h, apop osis,
EMT, mig a ion, in asion model [90]
221 CCA PTEN EMT, mig a ion, in asion Cell lines, issue [96]
224 CCA p15, p21, Cyclin E1 Cell cycle p og assion, umou g ow h Cell lines, issue [88]
421 CCA and gallbladde
cance FXR P oli e a ion, mig a ion Cell lines, issue [93]
429 CCA CDH6 Tumo g ow h Cell lines [91]
Cell lines, issue, se um
Cell lines, issue, animal
Cell lines, animal model
Cell lines, issue, animal
P oli e a ion, apop osis, EMT, mig a ion , in asion, oncogenesis,
P. Olaizola e al. BBA - Molecula Basis o Disease 1864 (2018) 1293–1307
1300
Tumo
supp esso
le -7c1CCA IL6, IL6R Mig a ion, in asion, in lamma ion, umo igenici y, s emness Cell lines, issue [112]
125b1CCA IL6R Mig a ion, in asion, in lamma ion, umo igenici y, s emness Cell lines, issue [112]
99a1CCA IGF1R Mig a ion, in asion, in lamma ion, umo igenici y, s emness Cell lines, issue [112]
26a CCA KRT19 P oli e a ion, umo g ow h model [120]
29a CCA HDAC4 Tumo g ow h, EMT, mig a ion, in asion Cell lines, issue [109]
29b CCA Mcl-1, PIK3R1, MMP2 Apop osis, chemo esis ance Cell lines [101, 118]
34a CCA c-Myc, SMAD4, c-Me , CDK6, Cyclin D1 Tumo g ow h, EMT, mig a ion, in asion, oncogenesis model [121-123]
122 CCA Cadhe in P oli e a ion, mig a ion, in asion model [100, 101]
138 CCA RhoC P oli e a ion, cell cycle p og ession, mig a ion, in asion Cell lines, issue [106]
145 iCCA NUAK P oli e a ion, umou g ow h, in asion Cell lines, issue [104]
150-5p CCA ELK1 P oli e a ion, umou g ow h, mig a ion, in asion [78]
199a-3p CCA mTOR Chemo esis ance Cell lines [119]
200c iCCA NCAM1 EMT, mig a ion, in asion Cell lines [107]
205 CCA -Chemo esis ance Cell lines [118]
212 iCCA FOXA1 P oli e a ion, in asion Cell lines, issue [103]
214 iCCA Twis EMT, mig a ion, in asion Cell lines, issue [105]
221 CCA PIK3R1 Apop osis, chemo esis ance Cell lines [118]
370 CCA MAP3K8, WNT10B P oli e a ion Cell lines, issue [113]
373 pCCA MBD2 Epigene ics Cell lines, issue [115]
376c iCCA GRB2 Mig a ion Cell lines [108]
410 CCA XIAP Tumo g ow h, apop osis, in asion Cell lines [103]
494 CCA CDK6, PTTG1, TOP2A P oli e a ion model [98, 99]
605 iCCA PSMD1/Ganky in P oli e a ion, apop osis, mig a ion, in asion, in lamma ion model [110]
Cell lines, issue, animal
Cell lines, issue, animal
Cell lines, issue, animal
Cell lines, issue, se um
Cell lines, issue, animal
Cell lines, issue, animal
Cyclin G1, IGF1R, MMP2, MMP9, RECK, E-Cadhe in, N-
1
miR clus e .
P. Olaizola e al. BBA - Molecula Basis o Disease 1864 (2018) 1293–1307
1301
