genes
G C A T
T A C G
G C A T
Re iew
The Po en ial o Zeb a ish as a Model O ganism o
Imp o ing he T ansla ion o Gene ic
An icance Nanomedicines
Gu ié ez-Lo e a C1,2,† ID , Vázquez-Ríos AJ2,†, Gue a-Va ela J1,3 ID , Sánchez L1and
de la Fuen e M2,*
1
Zoology, Gene ics and Physical An h opology Depa men Ve e ina y Facul y, Uni e sidade de San iago de
Compos ela, Lugo 27002, Spain; [email p o ec ed] (G.-L.C); jo [email p o ec ed] (G.-V.J);
[email p o ec ed] (S.L)
2Nano-Oncology Uni , T ansla ional Medical Oncology G oup, Heal h Resea ch Ins i u e o San iago de
Compos ela (IDIS), Clinical Uni e si y Hospi al o San iago de Compos ela (CHUS), CIBERONC,
San iago de Compos ela 15706, Spain; [email p o ec ed]
3Geneaqua S.L., Lugo 27002, Spain
*Co espondence: ma ia.de.la. uen e. ei e@se gas.es
† These au ho s con ibu ed equally o his wo k.
Academic Edi o : Paolo Cinelli
Recei ed: 9 Oc obe 2017; Accep ed: 21 No embe 2017; Published: 28 No embe 2017
Abs ac :
In he las ew decades, he ield o nanomedicine applied o cance has e olu ionized
cance ea men : se e al nano o mula ions ha e al eady eached he ma ke and a e ou inely being
used in he clinical p ac ice. In he case o gene ic nanomedicines, i.e., designed o deli e gene
he apies o cance cells o he apeu ic pu poses, ad ances ha e been less imp essi e. This is because
o he many ba ie s ha limi he access o he he apeu ic nucleic acids o hei a ge si e, and he
lack o models ha would allow o an imp o emen in he unde s anding o how nanoca ie s
can be ailo ed o o e come hem. Zeb a ish has impo an ad an ages as a model species o he
s udy o an icance he apies, and ha e a lo o o e ega ding he a ional de elopmen o e icien
deli e y o gene ic nanomedicines, and hence inc easing he chances o hei success ul ansla ion.
This e iew aims o p o ide an o e iew o he ecen ad ances in he de elopmen o gene ic
an icance nanomedicines, and o he zeb a ish models ha s and as p omising ools o shed ligh on
hei mechanisms o ac ion and o e all po en ial in oncology.
Keywo ds: nanomedicines; cance ; gene he apies; zeb a ish; ansla ion
1. Nano echnology P o ides Inno a i e App oaches o Cance Managemen
In ecen decades, an inc easing unde s anding o he molecula and biological basis o cance and
he disco e y o no el echnologies has led o imp o emen s in cance su i al. The de elopmen o
ea ly de ec ion ools and a ge ed ea men s, as well as changes in pa ien s’ li es yle, ha e con ibu ed
o his highe a e o cance su i al. The de elopmen o new nanomedicines o cance ea men is
an in e disciplina y esea ch ield ha includes biology, chemis y, enginee ing, and medicine, wi h a
clea goal: ad ancing cance de ec ion, diagnosis, and ea men .
Di e en ypes o nanoca ie s, including liposomes and o he lipid-based nanosys ems,
polyme -based nanopa icles, micelles, polyplexes, dend ime s, polyme somes and d ug/p o ein
conjuga es ha e been p oposed du ing he las ew decades in cance esea ch [
1
–
11
]. Fo cance
ea men , he goal is o enhance he e icacy and dec ease he oxici y o he cu en he apeu ics
by al e ing hei pha macokine ic p o ile, inc easing hei solubili y and s abili y in biological luids,
augmen ing hei accumula ion in umo s, and educing hei oxici y. Biological d ugs, such as gene
Genes 2017,8, 349; doi:10.3390/genes8120349 www.mdpi.com/jou nal/genes
Genes 2017,8, 349 2 o 20
he apies, pep ides and p o eins, can also bene i g ea ly om he applica ion o nano echnology
ha could p o ec hem om p ema u e deg ada ion and acili a e hei access o he in acellula
compa men [
12
–
15
]. Liposomes a e he mos common ype o nanos uc u e ha ha e ansla ed
in o ma ke ed p oduc s [
16
–
21
]. Back in 1995, he US Food and D ug Adminis a ion (FDA)
app o ed he i s nanopa icle o cance ea men , Doxil
©
, a liposomal nanopa icle loaded wi h
he chemo he apeu ic d ug doxo ubicin [
22
]. Since hen, o he nano he apeu ics based on liposomes
ha e eached he ma ke such as Pegyla ed liposomal doxo ubicin (Doxil
©
/Caelyx
©
), liposomal
cy a abine (DepoCy
©
), Dauno ubicin ci a e Liposomes (DaunoXome
©
), liposomal doxo ubicin
(Myoce
©
), Vinc is ine Sul a e Liposomes (Ma qibo
©
), liposomal i ino ecan (Oni yde
©
). Pacli axel
polyme ic nanopa icles (Opaxio
©
), pegyla ed L-aspa aginase polyme ic nanopa icles (Oncaspa
©
),
leup olide ace a e polyme ic micelles (Eliga d
©
), oxalipla in micelles (Eloxa in
©
), polyme –p o ein
conjuga e peg ilg as im (Neulas a
©
), albumin-pacli axel (Ab axane
©
), Denileukin di i ox (On ak
©
),
B en uximab-Monome hyl au is a in E (MMAE) (Adce is
™
), and T as uzumab-Em ansine (Kadcyla
©
)
a e examples o di e en ypes o nanos uc u es ha ha e led o p oduc s al eady in clinical use.
Apa om hei use in he possible de elopmen o nano he apeu ics, nanopa icles a e also
use ul ools in he diagnosis ield, due, in he case o ino ganic nanopa icles, o hei in insic
p ope ies ha allow a di ec acking, and, in he case o o ganic nanopa icles, o hei abili y
o accommoda e/encapsula e di e en molecules and con as agen s o imaging applica ions.
Many con as agen s a e cu en ly being s udied wi h his goal in mind, including supe -pa amagne ic
i on oxide nanopa icles and ul a-small supe -pa amagne ic i on oxide nanopa icles, hea y
me al (i.e., gold, lan hanide, and an alum) nanopa icles, echne ium-99m (
99m
TC) sulphu
colloid nanopa icles, I-labeled cRGDY silica nanopa icles, su ace-enhanced Raman sca e ing
nanopa icles, and single-walled ca bon nano ubes. O ganic nanopa icles such as liposomes, micelles,
and nanoemulsions can, o example, encapsula e supe -pa amagne ic i on oxide nanopa icles, o be
adiolabeled wi h adioiso opes such as
89
Z ,
111
In,
18
F,
64
Cu o
68
Ga o molecula imaging [
23
–
26
].
The imaging modali ies cu en ly a ailable expe imen ally a e: ul asound, magne ic esonance
imaging (MRI), op ical imaging, molecula imaging, compu ed omog aphy (CT), posi on emission
omog aphy (PET), and single-pho on emission compu ed omog aphy (SPECT). Howe e , in
clinics, he mos used modali ies o whole-body imaging a e CT, MRI, PET and SPECT. Fo
o gan-speci ic examina ions, ul asounds a e o p e e ence since hey a e as e and less expensi e,
while, o supe icial lesions, endoscopic, and in aope a i e p ocedu es, op ical and pho o-acous ic
applica ions a e mo e sui able [27,28].
Finally, nanopa icles also ha e a g ea po en ial as nano he anos ics, i.e., mul i unc ional
nanopa icles ha combine, in o a single en i y, elemen s o he apy and o diagnosis.
Nano he anos ics ha e been explo ed o applica ions combining di e en imaging modali ies
and he apeu ic applica ions, such as pho odynamic he apy, pho o he mal, pho o igge ed
chemo he apeu ic elease, ul asound igge ed, elec o- he mal, magne o he mal, X- ay,
and adio equency he apies [
27
,
29
]. Mo eo e , nano he anos ics a e ga he ing g ea in e es
because hey migh p o ide a deepe unde s anding o key aspec s ha could make a nanopa icle
o mula ion success ul—such as d ug elease kine ics and pene a ion o nanoca ie s wi hin
umo s—moni o ing he apeu ic esponses, as well as allowing he implemen a ion o no el s a egies,
such as imaging-guided local he apy [
30
,
31
]. To da e, he e is only one o mula ion unde going clinical
ials (Phase I) o he ea men o mul iple b ain me as ases, AGuIX
®
(Ac i a ion and Guidance
o I adia ion by X- ay), a gadolinium-based nanopa icle o a ound 5 nm diame e , de eloped
mainly o imaging applica ions due o i s magne ic esonance con as p ope ies. Howe e , when
i is combined wi h X- ay adia ion, i inc eases h ee- old he adio he apy e ec i eness in mice,
playing a double ole, as adiosensi ize and as imaging agen (NCT02820454) [
32
,
33
]. We belie e
ha nano he anos ics ha e a lo o po en ial in cance managemen , and could de ini i ely make an
impac in he clinical p ac ice by, concu en ly, diagnosing he disease, helping pa ien s s a i ica ion,
Genes 2017,8, 349 3 o 20
guiding ocal he apy, acking d ug elease and pene a ion wi hin umo s, moni o ing esponse, and,
i equi ed, swi ching ea men s.
2. Gene ic Nanomedicines and he Main Challenges o Thei T ansla ion o he Clinic
Ad ances in gene ics and molecula biology ha e led o he de elopmen o new he apies ha
can speci ically modula e he exp ession o ele an genes in o de o co ec abno mali ies and es o e
hei o iginal biological unc ion. Some o he s a egies o gene he apy include (i) silencing oncogene
exp ession, (ii) p omo ing umo -supp esso genes, (iii) co ec ing mu a ions, (i ) suicide gene he apy,
( ) supp essing umo angiogenesis, and ( i) ac i a ing an immune esponse agains umo cells.
Fo hese pu poses, plasmid DNA (pDNA), minici cles (supe coiled ci cula DNA), oligonucleo ides
(ASOs, decoys, ap ame s), RNA in e e ence (sho -hai pin (shRNA), small in e e ing RNA (siRNA)
and mic oRNA (miRNA)) a e being ex ensi ely explo ed [
34
]. Howe e , because naked nucleic acids
a e ulne able o enzyma ic deg ada ion, apid clea ance, and non-speci ic biodis ibu ion, only low
gene exp ession e iciencies can be achie ed. Hence, he p ima y challenge o gene he apy is o
de elop e ec i e ca ie s able o p o ec he nucleic acids and acili a e hei in e naliza ion in o he
a ge ed cells a he a ge ed si e [35].
T adi ionally, ec o s o gene he apy applica ions a e di ided in o i al and non- i al ca ie s.
Mos gene ec o s (~69%) cu en ly unde going clinical ials in ol e i uses (i.e., e o i uses,
len i i uses, adeno i uses, and adeno-associa ed i uses). In Augus 2017, he FDA app o ed
he i s gene he apy in he Uni ed S a es, Tisagenlecleucel (Kym iah
©
) om No a is Pha ma
AG (Basel, Swi ze land), o ce ain pedia ic and young adul pa ien s wi h a o m o acu e
lymphoblas ic leukemia whose i s -line d ugs ha e ailed [
36
]. This pionee gene he apy—based
on a sel -inac i a ing len i i al ec o ha con ains ex ensi ely modi ied sequences om HIV-1 so
as o deli e chime ic an igen ecep o (CAR)-encoding sequences in o T cells o a ge and kill
leukemia cells wi h speci ic an igen (CD19) on he su ace—achie es an o e all emission a e o
83% (52/63) in his pa ien popula ion [
37
]. Despi e hese ad ances, many conce ns s ill emain
ega ding he use o i al ec o s, such as hei po en ial immunogenici y, he possibili y o e e sion
o he i ulen o m o he i uses, and also hei high p oduc ion cos s [
35
]. Al e na i e syn he ic
ec o s, made ou o na u al, semi-syn he ic o syn he ic ma e ials, o e a sa e al e na i e o in oduce
gene ic ma e ials in o he a ge ed cells. Nume ous non- i al gene deli e y sys ems o di e en
ypes o nucleic acids (mainly pDNA, siRNA and miRNA) ha e been desc ibed o da e [
34
,
38
].
Di e en applica ions o he de elopmen o no el an icance gene ic nanomedicines ha e simila ly
being explo ed, including suicide gene he apies, an i-angiogenic gene he apies, immuno he apies,
es o a ion o oncosupp esso RNAs, o gene silencing o oncogenes, o speci ic non-coding RNAs
(an agomi s), o p o eins in ol ed in esis ance o chemo- and adio- he apies, an i-apop o ic p o eins,
epigene ic egula ion, e c., as ecen ly e iewed by Bo ai e al. [
39
]. The main p eclinical s udies o
he di e en applica ions o nanopa icles o gene he apy epo ed success ul in mice models a e
summa ized in Table 1( epo e genes and expe imen s e e ing o o e exp ession/silencing o
housekeeping genes a e no included).
Table 1.
Main s udies o da e o gene ic nanomedicines ha ha e had ele an he apeu ic e ec s on
di e en ypes o cance in mice models.
Nanoca ie Gene Vec o Ta ge Indica ion Adminis a ion
Rou e Re
Liposomes
miRNA Res o a ion o oncossupp esso B eas cance Tail ein [40]
siRNA EpCAM silencing B eas cance Tumo adjacen [41]
siRNA An i-angiogenesis B eas cance In a umo al [42]
miRNA Res o a ion o oncosupp esso Hepa ocellula
ca cinoma In a umo al [43]
shRNA WT1 silencing Melanoma Tail ein [44]
Genes 2017,8, 349 4 o 20
Table 1. Con .
Nanoca ie Gene Vec o Ta ge Indica ion Adminis a ion
Rou e Re
Polyme ic
nanopa icles
pDNA An i-angiogenesis Colon cance Tail ein [45]
pDNA Induce apop osis O a ian cance In ape i oneal [46]
pDNA Suicide gene he apy O a ian cance In ape i oneal [47]
pDNA Immuno he apy Colo ec al cance In a umo al [48]
pDNA Suicide gene he apy Colon cance In a umo al [49]
Lipid
nanopa icles
siRNA And ogen ecep o silencing P os a e cance Tail ein [50]
miRNA Res o a ion o mic oRNA-26a Lymphocy ic
leukemia In ape i oneal [51]
Dend ime s si/shRNA ITCH silencing Panc ea ic cance Tail ein [52]
siRNA, small in e e ence RNA; shRNA, sho -hai pin RNA; pDNA, plasmid DNA; miRNA, mic oRNA; EpCAM,
epi helial cell adhesion molecule; WT1, Wilms Tumo 1.
Recen ad ances in non- i al gene ec o s ega ding e iciency, speci ici y, sa e y and gene
exp ession du abili y ha e led o an inc ease in he numbe o nanopa icle-based gene deli e y ec o s
in clinical ials while he numbe o i al ec o s ha e d opped signi ican ly [
53
]. Some examples in
cance a e ela ed o liposomes o siRNA, mic oRNA o pDNA deli e y (NCT01591356, NCT01829971,
NCT01489371, NCT02340156); lipid nanopa icles (NCT02314052, NCT01437007) o polyme ic
nanopa icles (NCT02956317) [54]. Un o una ely, non- i al ec o s ha e no eached he ma ke ye .
The design o success ul syn he ic nano ec o s poses a big challenge since hey need o
o e come impo an biological ba ie s. Nano ec o s need (i) o be sa e and adequa e o pa en e al
adminis a ion, (ii) e icien ly p o ec nucleic acids om deg ada ion, and (iii) p omo e hei access
o he a ge in acellula compa men in he a ge cell (depending on he selec ed gene he apeu ic
sys em, i.e., plasmid DNA, RNAi, non-coding RNA (ncRNAs), oligonucleo ides, e c.), in enough
amoun s o media e a he apeu ic e ec (depending on he po ency o he molecule, speci ici y, and
s abili y) [
34
,
55
–
58
]. All hese aspec s should be aken in o conside a ion om ea ly de elopmen
o inc ease he chances o ansla ion in o ea ly-phase clinical ials [
11
,
59
–
61
]. The de elopmen o
unc ional assays and he selec ion o adequa e animal models o he apeu ic e alua ion a e also key
s eps ha c i ically a ec he ou come o he p eclinical e alua ion.
Al hough a numbe o gene-deli e y nano ec o s ha e been claimed o be e icien , mos
o he s udies ha e been done
in i o
, on immo alized cance cell lines, and only a ew
ha e ac ually add essed he he apeu ic ou come
in i o
. While
in i o
expe imen s include
e alua ion o oxici y (e.g., MTT (3-(4,5-Dime hyl hiazol-2-yl)-2,5-Diphenyl e azolium B omide),
MTS (3-(4,5-dime hyl hiazol-2-yl)-5-(3-ca boxyme hoxyphenyl)-2-(4-sul ophenyl)-2H- e azolium)
o ypan blue s aining assays), ans ec ion e iciency (e.g., in e naliza ion o luo escen
nanopa icles/nucleic acids by con ocal mic oscopy o /and low cy ome y), gene exp ession (e.g.,
RT-PCR, wes e n blo , o ELISA assays), and some imes unc ional assays (e.g., e alua ion o cell
p oli e a ion, mig a ion and in asion, colony o ma ion, angiogenesis, and apop osis),
in i o
epo s
in animal models (mainly oden s) a e mos ly limi ed o measu ing a he apeu ic e ec in e ms o
umo g ow h, p o iding only a yes o no answe . The e o e, he causes behind he he apeu ic ailu e
a e no well unde s ood. In ou opinion, i is necessa y o lea n mo e abou he
in i o
pe o mance
o gene ic nanomedicines, and o inco po a e unc ional assays in animal models, in o de o speed
up he ansla ion o gene ic nanomedicines o a clinical se ing. No el ools and models ha would
allow as and low-cos compa a i e s udies o he a ional op imiza ion o gene ic nanomedicines a e
u gen ly needed.
3. Zeb a ish as a Model Species
Zeb a ish (Danio e io) is a eshwa e ish belonging o he Cyp inidae amily, common in he
i e Ganga basin on he Indian sub-con inen . Zeb a ish has some well-known cha ac e is ics ha
makes i eally a ac i e as a model o human diseases [
62
–
65
]. In ac , i has achie ed he s a us
o model species, and been p esen ed as an ex ao dina y complemen o mu ine models, and a
Genes 2017,8, 349 5 o 20
p omising al e na i e [
64
]. Fo one, zeb a ish’s main enance is a o dable in e ms o easibili y and
cos s. Mo eo e , adul indi iduals a e small in size (2.5–4 cm), which makes he space equi emen s
no e y demanding. In addi ion, i has high ecundi y and e iliza ion a es (up o 200 e ilized eggs
pe ma ing pai and week), and p esen s ex e nal e iliza ion, which allows o pe o ming di ec ed
c osses, as well as
in i o
e iliza ion. I also p esen s ela i ely sho gene a ion imes—a ound h ee
mon hs. Finally, he genome o zeb a ish, whose comple e DNA sequence was published in 2013 [
66
],
shows app oxima ely 70% o homology wi h he human genome, and 82% o o hologous human
disease- ela ed genes.
Zeb a ish emb yos a e pa icula ly in e es ing o biomedical applica ions [
67
,
68
]. As ea ly as
48 h pos e iliza ion (hp ), emb yos aised a 28.5
◦
C ha ch om he cho ion (ex e nal and acellula
p o ec i e memb ane), and become ee-li ing animals wi h a comple e body pa e n, and almos
comple ely unc ional o gans [
69
]. A his ime, he inna e immune sys em is al eady ac i e [
70
],
bu he adap i e immune sys em will no be ully ope a ing un il 4–6 weeks pos e iliza ion (wp ) [
71
],
al hough exp ession o some genes o he adap i e immune sys em s a s as ea ly as eigh days pos
e iliza ion (dp ) [
72
]. The e o e, he esul s o analyses ca ied ou du ing he emb yo-la al phases
can be aced back o he inna e immune sys em.
Zeb a ish emb yos a e obus and can su i e di e en p ocedu es igh a e e iliza ion,
including gene ic manipula ion, mo pholino [
73
–
75
] o ibonucleop o ein (CRISPR/Cas) [
76
–
79
]
mic oinjec ion a single cell s age, as well as cance cell xeno ansplan s [
80
–
85
]. In addi ion, hey a e
anspa en , which gi es hem a de ini e ad an age in many ields o s udy, because i makes
possible, o example, o examine he de elopmen o in e nal s uc u es, and he acking o he
mo emen s and biodis ibu ion o labeled pa icles (mic oo ganisms, cells, nanopa icles
. . .
) in eal
ime [
85
–
88
]. Visualiza ion can be hampe ed by he ea ly p oduc ion o melanin du ing hei emb yonic
de elopmen , as ea ly as 24 hp (p im5 de elopmen al s age). Howe e , melanin p oduc ion can be
easily blocked by ea ing he emb yos wi h 1-phenyl 2- hiou ea (PTU) [
69
]. Addi ionally, he small
size o zeb a ish emb yos (assays can be pe o med in 96 o , less sui ably, in 384 mul i-well pla es),
and he ac ha hey can li e in small olumes (so ha low quan i ies o he es ed compounds a e
equi ed) make his a sui able model o high- h oughpu analyses [
89
,
90
]. An adul also anspa en
line (caspe ) was de eloped [91], which allows o ca ying ou simila analyses in adul s [92–94].
Finally, he Eu opean Food Sa e y Adminis a ion [
95
] has s a ed ha ish in hese ea ly
de elopmen al s ages, up o 5 dp , a e less likely o expe ience pain, su e ing, dis ess, o su e
las ing ha m, in acco dance wi h he 3Rs P inciples ( eplacemen , educ ion, and e inemen ) o
humane animal esea ch [96].
The e o e, aking all hese ac s in o conside a ion, zeb a ish has been accep ed as a sui able model
o biomedical pu poses, o i could p o ide esul s as e han esea ch on non- anspa en , less
p oli ic, mo e ime-consuming, and expensi e oden s, and imp o e he biological in e p e a ion o he
esul s compa ed o wo king on in e eb a e models, which a e phylogene ically u he om human
beings, and om in i o analyses, which lack body in e ac ions.
4. Zeb a ish Is Cu en ly Being Used o he De elopmen o An icance The apeu ics
The pa hological mechanisms unde lying cance a e some o he mos challenging p ocesses o
unde s and because o hei a ie y and complexi y. Zeb a ish is conside ed a complemen a y model
o mu ine and o he p e ious models o he s udy o he gene ic basis o cance and o he e alua ion
o ca cinogenic and no el an i umo al compounds in d ug disco e y [97–108].
Zeb a ish has p o en o be a good model o p edic ad e se d ug e ec s du ing animal p eclinical
and human clinical da a [
109
]. This is because many o he cellula and molecula mechanisms in ol ed
in zeb a ish’s esponse o oxici y o s ess a e simila o hose o mammals [
110
,
111
]. The publica ion
o he DNA sequence o he zeb a ish genome con i med ha ele an molecula pa hways, including
hose implica ed in cance , a e simila o hose o mammals [
66
], which made zeb a ish an a ac i e
choice o cance esea ch [
67
,
106
,
112
,
113
]. A pa allel app oach o modeling cance has been he (xeno)
Genes 2017,8, 349 6 o 20
ansplan o human cance cells in o zeb a ish emb yos, which led o he de elopmen o he so-called
xenog a ed emb yos. The p oli e a ion, sp eading and me as asizing o mic oinjec ed cance cells is
possible because he zeb a ish emb yos lack an adap i e immune sys em. Since he i s success ul
model in 2005 and u he imp o emen s in 2006 [
80
,
114
], di e en xenog a zeb a ish models ha e
been epo ed bea ing ei he comme cial human cance cell lines o p ima y umo cells, including
cance s om di e en o igins (i.e., melanoma, b eas ca cinoma, colo ec al, panc ea ic, o a ian, kidney,
lung, o al, p os a e, leukemia, e c.) [80,82,85,115–119].
As indica ed abo e, zeb a ish cance models ha e been used o no el d ug sc eening, as well as
o eanalysis o known d ugs [
97
,
100
,
105
–
108
,
113
,
120
]. Ne e heless, due o he nano echnology
e olu ion on an icance d ug deli e y, as s a ed in Sec ion 1, ecen s udies also highligh he
po en ial o zeb a ish o he e alua ion o no el an icance nanomedicines. Mos s udies measu ed he
oxici y and sa e y o blank nanopa icles (i.e., p io o d ug inco po a ion) using di e en p ocedu es,
bu also co e ed mo phological desc ip ions o zeb a ish a e adminis a ion o sub le hal doses,
and expe imen s o gene exp ession [
68
,
121
–
123
]. Taking ad an age o he emb yo anspa ency,
biodis ibu ion s udies ha e also been pe o med o de e mine he abili y o he nanoca ie s o each
he a ge si e, and e en su pass complex biological ba ie s, such as he blood–b ain ba ie [
124
–
126
].
Apa om de e mining hese c i ical pa ame e s, he zeb a ish xenog a model has also been
p o en use ul in he s udy o he in e ac ion be ween d ug-loaded nanoca ie s and xenog a ed
cells, o example when s udying a possible educ ion in he popula ion o cance cells [
68
,
127
–
129
].
Among o he s, i is wo h men ioning Yang and collabo a o s’ s udies [
129
] ha desc ibe he in e ac ion
o a ge ed doxo ubicin-loaded liposomes wi h HeLa cells, and he e iciency o his s a egy in a
xenog a model o zeb a ish, and also he wo k o E ensen and collabo a o s [
127
] ha desc ibes he
abili y o PEGyla ed nanoca ie s o a oid up ake by mac ophages, a ac ha ansla es in imp o ed
ci cula ion imes and inc eased accumula ion in o he umo s. Figu e 1depic s a isual example
o liposomes labeled in g een and dis ibu ed along he ish blood essels upon injec ion in o he
ci cula ion (A) and hei subsequen up ake by mac ophages labeled in ed (yellow do s).
Genes 2017, 8, 349 6 o 20
he oxici y and sa e y o blank nanopa icles (i.e., p io o d ug inco po a ion) using di e en
p ocedu es, bu also co e ed mo phological desc ip ions o zeb a ish a e adminis a ion o sub
le hal doses, and expe imen s o gene exp ession [68,121–123]. Taking ad an age o he emb yo
anspa ency, biodis ibu ion s udies ha e also been pe o med o de e mine he abili y o he
nanoca ie s o each he a ge si e, and e en su pass complex biological ba ie s, such as he blood–
b ain ba ie [124–126]. Apa om de e mining hese c i ical pa ame e s, he zeb a ish xenog a
model has also been p o en use ul in he s udy o he in e ac ion be ween d ug-loaded nanoca ie s
and xenog a ed cells, o example when s udying a possible educ ion in he popula ion o cance
cells [68,127–129]. Among o he s, i is wo h men ioning Yang and collabo a o s’ s udies [129] ha
desc ibe he in e ac ion o a ge ed doxo ubicin-loaded liposomes wi h HeLa cells, and he e iciency
o his s a egy in a xenog a model o zeb a ish, and also he wo k o E ensen and collabo a o s
[127] ha desc ibes he abili y o PEGyla ed nanoca ie s o a oid up ake by mac ophages, a ac ha
ansla es in imp o ed ci cula ion imes and inc eased accumula ion in o he umo s. Figu e 1 depic s
a isual example o liposomes labeled in g een and dis ibu ed along he ish blood essels upon injec ion
in o he ci cula ion (A) and hei subsequen up ake by mac ophages labeled in ed (yellow do s).
Figu e 1. G een-labeled liposomes, injec ed in o he ci cula o y sys em o wild ype zeb a ish emb yos
(A), allows he isualiza ion o he luo escen liposomes in he ish ascula u e. On he igh , he g
(mpeg1meche y) model (B) shows he up ake o he luo escen g een liposomes by luo escen ed
ci cula ing mac ophages (yellow do s). Imaging adap ed om he wo k o E ensen e al. [127] wi h
pe mission.
5. The Po en ial o Zeb a ish o Inc easing he T ansla ion o Gene ic An icance
Nanomedicines: Ba ie s and Models
Apa om he use o zeb a ish o he de elopmen o no el cance he apeu ics, including
nano he apeu ics, only a ew s udies ha e been epo ed using his model o es p eclinical gene ic
nanomedicines [130–132]. The i s s udy ound in he li e a u e e alua es a syne gis ic he apy based
on he co-encapsula ion o a pigmen -epi helium-de i ed ac o (PEDF) plasmid wi h pacli axel, a
small molecula chemo he apeu ic d ug, in o poly(lac ic-co-glycolic acid) (PLGA) nanopa icles, in a
ansgenic zeb a ish model Flk-1:eGFP. The esul s showed an ac i e a ge ing ha ansla es in o an
e ec i e and sa e an iangiogenic he apy [130]. The second example co e s he de elopmen o a
e o-in e se amphipa hic RICK ( e o-in e se o m o he CADY-K pep ide) pep ide as no el non-
co alen siRNA ca ie . The designed nanopa icles show an e ec i e siRNA p o ec ion, based on
he speci ic p o ease esis an pep ide sequence. The au ho s in es iga ed he e ec o a polye hylene
glycol (PEG) g a ing o RICK nanopa icles on hei in i o and in i o capaci y o deli e siRNA.
In i o assays pe o med in Caspe zeb a ish ollowed he biodis ibu ion o luo escen -labeled
nanopa icles a e injec ion a he one-cell s age in zeb a ish emb yos. The au ho s desc ibed a
modula , easy- o-handle d ug deli e y sys em ha could be adap ed o o he ypes o unc ional
moie ies in o de o de elop sa e and biocompa ible deli e y sys ems o he clinical applica ion o
RNAi-based cance he apeu ics [131]. Finally, Co dei o e al. [132] epo ed he design o a gold
nanobeacon able o silence enhanced g een luo escence p o ein (EGFP) in emb yos o a li-EGFP
ansgenic zeb a ish line. Resul s in his model allowed he au ho s o conclude ha hey ha e
de eloped a biocompa ible and e icien nanopla o m o gene silencing pu poses.
As illus a ed in Figu e 2, a close e alua ion o he in i o pe o mance o gene ic
nanomedicines and a de ailed s udy o hei abili y o o e come he c i ical ba ie s ha migh
hampe a success ul he apy a e key ac o s in o de o speed up hei ansla ion o clinic.
Figu e 1.
G een-labeled liposomes, injec ed in o he ci cula o y sys em o wild ype zeb a ish emb yos
(
A
), allows he isualiza ion o he luo escen liposomes in he ish ascula u e. On he igh , he g
(mpeg1meche y) model (
B
) shows he up ake o he luo escen g een liposomes by luo escen
ed ci cula ing mac ophages (yellow do s). Imaging adap ed om he wo k o E ensen e al. [
127
]
wi h pe mission.
5. The Po en ial o Zeb a ish o Inc easing he T ansla ion o Gene ic An icance Nanomedicines:
Ba ie s and Models
Apa om he use o zeb a ish o he de elopmen o no el cance he apeu ics, including
nano he apeu ics, only a ew s udies ha e been epo ed using his model o es p eclinical gene ic
nanomedicines [
130
–
132
]. The i s s udy ound in he li e a u e e alua es a syne gis ic he apy based
on he co-encapsula ion o a pigmen -epi helium-de i ed ac o (PEDF) plasmid wi h pacli axel, a small
molecula chemo he apeu ic d ug, in o poly(lac ic-co-glycolic acid) (PLGA) nanopa icles, in a ansgenic
zeb a ish model Flk-1:eGFP. The esul s showed an ac i e a ge ing ha ansla es in o an e ec i e
and sa e an iangiogenic he apy [
130
]. The second example co e s he de elopmen o a e o-in e se
amphipa hic RICK ( e o-in e se o m o he CADY-K pep ide) pep ide as no el non-co alen siRNA
ca ie . The designed nanopa icles show an e ec i e siRNA p o ec ion, based on he speci ic p o ease
esis an pep ide sequence. The au ho s in es iga ed he e ec o a polye hylene glycol (PEG) g a ing o
RICK nanopa icles on hei
in i o
and
in i o
capaci y o deli e siRNA.
In i o
assays pe o med in
Genes 2017,8, 349 7 o 20
Caspe zeb a ish ollowed he biodis ibu ion o luo escen -labeled nanopa icles a e injec ion a he
one-cell s age in zeb a ish emb yos. The au ho s desc ibed a modula , easy- o-handle d ug deli e y sys em
ha could be adap ed o o he ypes o unc ional moie ies in o de o de elop sa e and biocompa ible
deli e y sys ems o he clinical applica ion o RNAi-based cance he apeu ics [
131
]. Finally, Co dei o
e al. [
132
] epo ed he design o a gold nanobeacon able o silence enhanced g een luo escence p o ein
(EGFP) in emb yos o a li-EGFP ansgenic zeb a ish line. Resul s in his model allowed he au ho s o
conclude ha hey ha e de eloped a biocompa ible and e icien nanopla o m o gene silencing pu poses.
As illus a ed in Figu e 2, a close e alua ion o he
in i o
pe o mance o gene ic nanomedicines
and a de ailed s udy o hei abili y o o e come he c i ical ba ie s ha migh hampe a success ul
he apy a e key ac o s in o de o speed up hei ansla ion o clinic.
Nex , we desc ibe he mos ele an ba ie s o gene deli e y, and he zeb a ish models ha , in ou
unde s anding, can be use ul o a a ional design o success ul an icance gene ic nanomedicines (compiled
in Table 2).
Genes 2017, 8, 349 7 o 20
Nex , we desc ibe he mos ele an ba ie s o gene deli e y, and he zeb a ish models ha , in
ou unde s anding, can be use ul o a a ional design o success ul an icance gene ic nanomedicines
(compiled in Table 2).
Figu e 2. Zeb a ish as a model o ganism o p eclinical s udies o gene ic nanomedicines. This scheme
highligh s he main cha ac e is ics o zeb a ish as model o ganisms and he main ad an ages o
nanomedicines o gene deli e y. The scope o his e iew is summa ized in he lowe sec ion o he
igu e whe e we ha e illus a ed di e en ways in which zeb a ish models can be ex emely use ul o
help us unde s and he biological beha iou o gene ic nanomedicines, and de ine be e p o o ypes
wi h imp o ed oppo uni ies o ansla ion o a clinical se ing. Zeb a ish models would allow
pe o ming se e al assays o in e es such as (i) e alua ion o he oxicological p o ile, (ii)
de e mina ion o he s abili y and hal -li e ci cula ion o nanomedicines inyec ed in he ish ci cula ion
sys em, (iii) s udy o he abili y o nanomedicines o ex a asa e, di use, pene a e in o he umo , and
in e ac wi h he a ge ed cells, and (i ) unc ional assays o es he po en ial and he e icacy o he
p oposed nanomedicines. The wo images on op co espond o a zeb a ish emb yo (le ), and o
nanome ic (~100 nm) lipidic nanoemulsions obse ed by a omic o ce mic oscopy (AFM) ( igh ).
Images in he low pa o he igu e co espond, om le o igh , o 48 hp mal o med zeb a ish
emb yo due o oxic e ec s o nanocapsules (image ep oduced wi h pe mission om Teijei o-Valiño
e al. [88], luo escen DiD-labelled lipidic nanoemulsions (blue) injec ed in o he ish ci cula ion
sys em and obse ed unde a luo escence mic oscope (images adqui ed a 48 h pos -injec ion),
luo escen nanopa icles ( ed) able o ex a asa e blood essels (g een) in a zeb a ish model (image
ob ained by con ocal mic oscopy by Zou e al. [133], and ep oduced wi h pe mission), and
luo escen DiD-labelled lipidic nanoemulsions ( ed) able o in e ac wi h cance cells (g een) in
xeno ansplan ed zeb a ish emb yos (HCT116-GFP) a e yolk mic oinjec ion.
5.1. Toxici y
Despi e he abili y o he nanopa icles o educe he side e ec s o he associa ed d ugs, ad e se
e ec s due o he nanopa icles hemsel es ha e been epo ed in some clinical s udies, including
immuno oxici y (alle gy, hype -sensi i i y, and immunosupp ession), acu e oxici y (i.e., single-dose
s udies), subacu e oxici y (i.e., epea ed-dose s udies o semi-ch onic oxici y s udies),
ca cinogenici y, ep oduc i e oxici y, de elopmen al oxici y, geno oxici y, hepa o oxici y o
epigeno oxici y [134–139]. Nanopa icles may also ac i a e inna e immuni y esponses in he body
and, as a consequence, hey can media e an uncon olled deli e y o p o-in lamma o y media o s
(anaphyla oxins) ha could nulli y he he apeu ic e ec o he nanoca ie and, e en wo se, p omo e
umo g ow h [140]. In he case o gene ic nanomedicines, hey ypically con ain ca ionic elemen s o
Figu e 2.
Zeb a ish as a model o ganism o p eclinical s udies o gene ic nanomedicines. This scheme
highligh s he main cha ac e is ics o zeb a ish as model o ganisms and he main ad an ages o
nanomedicines o gene deli e y. The scope o his e iew is summa ized in he lowe sec ion o he
igu e whe e we ha e illus a ed di e en ways in which zeb a ish models can be ex emely use ul o
help us unde s and he biological beha iou o gene ic nanomedicines, and de ine be e p o o ypes
wi h imp o ed oppo uni ies o ansla ion o a clinical se ing. Zeb a ish models would allow
pe o ming se e al assays o in e es such as (i) e alua ion o he oxicological p o ile, (ii) de e mina ion
o he s abili y and hal -li e ci cula ion o nanomedicines inyec ed in he ish ci cula ion sys em,
(iii) s udy o he abili y o nanomedicines o ex a asa e, di use, pene a e in o he umo , and in e ac
wi h he a ge ed cells, and (i ) unc ional assays o es he po en ial and he e icacy o he p oposed
nanomedicines. The wo images on op co espond o a zeb a ish emb yo (le ), and o nanome ic
(~100 nm) lipidic nanoemulsions obse ed by a omic o ce mic oscopy (AFM) ( igh ). Images in he
low pa o he igu e co espond, om le o igh , o 48 hp mal o med zeb a ish emb yo due o oxic
e ec s o nanocapsules (image ep oduced wi h pe mission om Teijei o-Valiño e al. [
88
], luo escen
DiD-labelled lipidic nanoemulsions (blue) injec ed in o he ish ci cula ion sys em and obse ed unde
a luo escence mic oscope (images adqui ed a 48 h pos -injec ion), luo escen nanopa icles ( ed) able
o ex a asa e blood essels (g een) in a zeb a ish model (image ob ained by con ocal mic oscopy by
Zou e al. [
133
], and ep oduced wi h pe mission), and luo escen DiD-labelled lipidic nanoemulsions
( ed) able o in e ac wi h cance cells (g een) in xeno ansplan ed zeb a ish emb yos (HCT116-GFP)
a e yolk mic oinjec ion.
Genes 2017,8, 349 8 o 20
5.1. Toxici y
Despi e he abili y o he nanopa icles o educe he side e ec s o he associa ed d ugs, ad e se
e ec s due o he nanopa icles hemsel es ha e been epo ed in some clinical s udies, including
immuno oxici y (alle gy, hype -sensi i i y, and immunosupp ession), acu e oxici y (i.e., single-dose
s udies), subacu e oxici y (i.e., epea ed-dose s udies o semi-ch onic oxici y s udies), ca cinogenici y,
ep oduc i e oxici y, de elopmen al oxici y, geno oxici y, hepa o oxici y o epigeno oxici y [
134
–
139
].
Nanopa icles may also ac i a e inna e immuni y esponses in he body and, as a consequence, hey can
media e an uncon olled deli e y o p o-in lamma o y media o s (anaphyla oxins) ha could nulli y
he he apeu ic e ec o he nanoca ie and, e en wo se, p omo e umo g ow h [
140
]. In he case
o gene ic nanomedicines, hey ypically con ain ca ionic elemen s o imp o e hei associa ion wi h
he anionic nucleic acids. These posi i ely cha ged bioma e ials ha e also been ela ed o oxici y and
o - a ge unspeci ic e ec s a e ans ec ion. Toxici y in p eclinical s udies elies mainly on simple
and con en ional es s (e.g., MTT assay), and, in some cases, sys emic oxici y
in i o
(e.g., se ological
and biochemical analysis o blood samples in mice). The e o e, i is clea ha oxici y needs u he
a en ion be o e we can p oceed o clinical s udies.
As men ioned in Sec ion 4, zeb a ish is widely used o he e alua ion o he ad e se e ec s
o d ugs, and o de e mine he ac i i y o an i umo compounds [
97
–
108
]. I could also be used o
de e mine he p eclinical oxici y o nanoca ie s o gene deli e y pu poses. The mos common and
simple oxici y s udies in wild ype zeb a ish ela e o acu e and ch onic e ec s. P o ocols o hese
s udies ha e al eady been app o ed by he O ganiza ion o Economic Co-ope a ion and De elopmen
(OECD). To de e mine zeb a ish emb yo oxici y, pos e iliza ion emb yos a e placed in a s a ic pla e
and exposed o he compound. The a e o mo phological changes is one o he endpoin s used o
gene a e dose esponse cu es [
141
,
142
]. The oxici y o se e al ypes o nanopa icles, mainly ino ganic
nanopa icles, has al eady been de e mined in zeb a ish using his es [
68
,
104
,
127
,
143
,
144
]. One
impo an pa ame e o oxici y e alua ion is he ha ching e iciency because nanopa icles can in e ac
wi h ha ching enzymes [
145
]. Zeb a ish is also a e sa ile o ganism o geno oxici y s udies [
146
–
149
],
de elopmen al and beha io al analysis [
150
–
153
], immuno oxici y [
154
,
155
], neu o oxici y [
156
,
157
],
and ep oduc i e oxici y s udies [
158
]. Fo example, in expe imen s wi h ansgenic lines, such as
Tg( lk1:eGFP), Tg(cmlc2:eGFP), Hsp70:eGFP, ARE:eGFP, FLI-1, and Nac e/ li1:EGFP, i was possible
o obse e he chemical-induced oxici y o nanocomposi es and me al oxide nanopa icles in eal
ime [159–163].
Zeb a ish is also an excellen model o p o ide no el insigh s on he in e ac ion be ween he
immune sys em and umo cells [
164
,
165
]. Because in zeb a ish, mac ophages play an impo an
ole in angiogenesis, his model could also be used o de elop unc ional assays ela ed o he
angiogenic p ocess (Sec ion 5.4). A ansgenic zeb a ish line, mpo:GFP, which exp esses GFP unde he
neu ophil-speci ic myelope oxidase p omo e , has also been desc ibed and used o s udy neu ophil
esponse [
166
], including he e alua ion o oxida i e s ess and in lamma o y esponses in neu ophils
ollowing he adminis a ion o silica nanopa icles [
167
]. In addi ion, s udies ega ding ca dio oxici y
a e also o g ea impo ance, among hem is wo h men ioning he e alua ion o e ec s occu ing
immedia ely a e adminis a ion and hei consequences [153].
5.2. S abili y and Hal -Li e While in Ci cula ion
P eclinical s udies some imes igno e he ac ha he elec os a ic s abili y o nanoca ie s
in i o
does no gua an ee hei s abili y
in i o
. Mo eo e , in many cases, he nanoca ie and he gene
ec o a e associa ed by elec os a ic in e ac ions. Upon con ac wi h a biological media o high ionic
s eng h, his sys em may agg ega e, esul ing in he displacemen o he nucleic acids ha could
be p ema u ely eleased in o he ci cula ion be o e eaching he a ge cells. The p esence o se um
p o eins (e.g., glycosaminoglycans) could ha e he same e ec . The e o e, a ho ough s udy, ele an
in i o
models, o he s abili y and in e ac ions o he nanoca ie unde s udy could be necessa y o
ensu e ha he associa ed nucleic acids a e no p ema u ely eleased in o he ci cula ion [
168
–
170
].
Genes 2017,8, 349 9 o 20
On he o he hand, nanosys ems should also be able o a oid ecogni ion by mac ophages, and a apid
clea ance by he mononuclea phagocy e sys em (MPS), which would lead o hei as emo al om
ci cula ion [136].
As s a ed in Sec ion 2, one o he main ad an ages o zeb a ish emb yos and adul s om he Caspe
line is ha hey a e anspa en , and he e o e sui able o di ec and eal- ime acking o luo escen
nanopa icles in o he ish ci cula ion, using high- esolu ion con ocal mic oscopy [
91
]. Impo an ly,
a ecen s udy shows a good co ela ion among pha macokine ic da a ob ained in zeb a ish, a ,
and mice, and highligh s he po en ial o zeb a ish o his pu pose [
126
]. Di e en s udies ca ied
ou wi h model nanopa icles, FluoSphe es
®
and Quan um Do s
®
, highligh he in luence o he
exposu e ou e (wa e bo ne, injec ion and o al), and su ace p ope ies o he nanopa icles on hei
biodis ibu ion and umo up ake [35,171].
One model use ul o acking he ci cula ion o nanopa icles is he ansgenic line Fli1:eGFP [
127
].
This line has allowed o ollowing he dis ibu ion and umo accumula ion o PEGyla ed
nanopa icles. In he same s udy, he Tg(mpeg1:mChe y) line was selec ed o e alua e he in e ac ion
o hese nanopa icles wi h mac ophages, which led o he conclusion ha PEG coa ing ac ually
dec eased he in e ac ion o he nanopa icles wi h mac ophages. T ansgenic lines o mac ophages,
neu ophils, and endo helial cells exp essing luo escen ma ke s (see Table 2) ha e also been used o
wa ch he in e ac ion be ween lipid nanopa icles and immune cells [87].
5.3. Ex a asa ion, Pene a ion in o he Tumo , and In e ac ion wi h he Ta ge Cells
Nanoca ie s should be able o exi he sys emic ci cula ion a he ac ion si e. Recen ly, i has
been epo ed ha cu en animal models ail o p edic he accumula ion o nanoca ie s inside he
umo , which is ac ually abou 0.7% o he injec ed dose [
172
,
173
]. Thus, animal models ha would
allow us o be e s udy he abili y o nanoca ie s in his s ep a e c ucial o ensu ing an e ec i e
he apeu ic e ec [
174
]. The complexi y o he umo ex acellula ma ix (ECM) may also es ic
he ex a asa ion o he nanoca ie s. Addi ionally, e en i he nanoca ie s could c oss he umo
ascula u e, hey migh no be able o pene a e deep enough inside he umo mass due o he high
in e s i ial luid p essu e, and migh accumula e ins ead in he pe iphe al a eas, o in he su ounding
heal hy issue [
15
,
175
]. Finally, he nanopa icles need o in e ac wi h he a ge cells. Typically,
he apies a e di ec ed a umo cells, bu hey can also be designed o a ge cells o he s oma o o
in il a e immune cells, cance s em cells (CSCs), cance -associa ed ib oblas s (CAFs), umo -associa ed
mac ophages (TAMs), pe icy es, endo helial cells, e c. [15,176].
To da e, an ex ensi e lis o imp o ed zeb a ish cance models has been epo ed, including
models o s udy neu oblas oma, b ain cance , eye cance , leukemia, melanoma, u eal melanoma,
and li e cance , among o he s [
177
]. Mo e complex models o s udy he mechanisms o umo cell
dissemina ion and me as ases o ma ion ha e also been epo ed [
178
,
179
]. Fo example, he model
Flk1:EGFP has been used o s udy he me as a ic sp ead a e injec ion o ed luo escen p o ein
(RFP)-labeled Hela cells in he caudal a e y [
180
]. O he esul s show how me as a ic cell lines ha e
imp o ed abili ies o mig a e and p oli e a e compa ed o cells isola ed om p ima y umo s [
181
].
The s udy o CSC has also been conside ed in zeb a ish models [
182
,
183
]. Rega ding he s udy o
he umo mic oen i onmen (TME), Zhao e al. [
184
] showed ha ans o ming g ow h ac o be a
(TGF-
β
) induced a p o- umo neu ophil cy okine exp ession pa e n in zeb a ish, and concluded ha
essen ial mechanisms in he cons i u ion o he TME a e conse ed in his model.
Rega ding he pa icula e alua ion o nanomedicines, se e al wo ks co e he e alua ion o
hei abili y o accumula e in umo cells a e injec ion in zeb a ish xenog a s [
125
,
127
,
185
–
187
].
Zeb a ish can he e o e be conside ed as a dynamic model o s udy he anspo and accumula ion
o nanopa icles.
Genes 2017,8, 349 16 o 20
103.
Lenis-Rojas, O.A.; Fe nandes, A.R.; Roma-Rod igues, C.; Bap is a, P.V.; Ma ques, F.; Pé ez-Fe nández, D.;
Gue a-Va ela, J.; Sánchez, L.; Vázquez-Ga cía, D.; To es, M.L.; e al. He e olep ic mononuclea compounds
o u henium(ii): syn hesis, s uc u al analyses,
in i o
an i umo ac i i y and
in i o
oxici y on zeb a ish
emb yos. Dal on T ans. 2016,45, 19127–19140. [C ossRe ] [PubMed]
104.
Lenis-Rojas, O.A.; Roma-Rod igues, C.; Fe nandes, A.R.; Ma ques, F.; Pé ez-Fe nández, D.; Gue a-Va ela, J.;
Sánchez, L.; Vázquez-Ga cía, D.; López-To es, M.; Fe nández, A.; e al. Dinuclea RuII(bipy)2 De i a i es:
S uc u al, Biological, and in Vi o Zeb a ish Toxici y E alua ion. Ino g. Chem.
2017
,56, 7127–7144. [C ossRe ]
[PubMed]
105.
Penas, C.; Sánchez, M.I.; Gue a-Va ela, J.; Sanchez, L.; Vázquez, M.E.; Masca eñas, J.L. Ligh -Con olled
Cellula In e naliza ion and Cy o oxici y o Nucleic Acid-Binding Agen s: S udies in Vi o and in Zeb a ish
Emb yos. ChemBioChem 2016,17, 37–41. [C ossRe ] [PubMed]
106.
Blackbu n, J.S.; Langenau, D.M. Zeb a ish as a model o assess cance he e ogenei y, p og ession and elapse.
Dis. Model. Mech. 2014,7, 755–762. [C ossRe ] [PubMed]
107.
Ta , J.; Liu, M.; Wen, X.Y. Zeb a ish cance and me as asis models o
in i o
d ug disco e y. D ug Disco .
Today Technol. 2013,10, e83–e89. [C ossRe ] [PubMed]
108.
Veino e, C.J.; Dellai e, G.; Be man, J.N. Hooking he big one: The po en ial o zeb a ish xeno ansplan a ion
o e o m cance d ug sc eening in he genomic e a. Dis. Model Mech.
2014
,7, 745–754. [C ossRe ] [PubMed]
109.
Be ghmans, S.; Bu le , P.; Goldsmi h, P.; Wald on, G.; Ga dne , I.; Golde , Z.; Richa ds, F.M.; Kimbe , G.;
Roach, A.; Alde on, W.; e al. Zeb a ish based assays o he assessmen o ca diac, isual and gu
unc ion—po en ial sa e y sc eens o ea ly d ug disco e y. J. Pha macol. Toxicol. Me hods
2008
,58, 59–68.
[C ossRe ] [PubMed]
110.
Yang, L.; Kemadjou, J.R.; Zinsmeis e , C.; Baue , M.; Leg adi, J.; Mülle , F.; Pank a z, M.; Jäkel, J.; S ähle, U.
T ansc ip ional p o iling e eals ba code-like oxicogenomic esponses in he zeb a ish emb yo. Genome Biol.
2007,8, R227. [C ossRe ] [PubMed]
111.
Simmons, S.O.; Fan, C.Y.; Ramabhad an, R. Cellula s ess esponse pa hway sys em as a sen inel ensemble
in oxicological sc eening. Toxicol. Sci. 2009,111, 202–225. [C ossRe ] [PubMed]
112.
Ba bazuk, W.B.; Ko , I.; Kada i, C.; Heyen, J.; Ta e, S.; Wun, E.; Bedell, J.A.; McPhe son, J.D.; Johnson, S.L.
The syn henic ela ionship o he zeb a ish and human genomes. Genome Res.
2000
,10, 1351–1358. [C ossRe ]
[PubMed]
113.
Hui ing, L.N.; La oche, F.; Feng, H. The Zeb a ish as a Tool o Cance D ug Disco e y. Aus in J. Pha macol. The .
2015,3, 1069. [PubMed]
114.
Lee, L.M.J.; Se o , E.A.; Bonde, G.; Co nell, R.A.; Hend ix, M.J.C. The a e o human malignan melanoma
cells ansplan ed in o zeb a ish emb yos: Assessmen o mig a ion and cell di ision in he absence o umo
o ma ion. De . Dyn. 2005,233, 1560–1570. [C ossRe ] [PubMed]
115.
Ma ques, I.J.; Weiss, F.U.; Vlecken, D.H.; Ni sche, C.; Bakke s, J.; Lagendijk, A.K.; Pa ecke, L.I.;
Heidecke, C.-D.; Le ch, M.M.; Bagowski, C.P. Me as a ic beha iou o p ima y human umou s in a zeb a ish
xeno ansplan a ion model. BMC Cance 2009,9, 128. [C ossRe ] [PubMed]
116.
Bansal, N.; Da is, S.; Te eshchenko, I.; Budak-alpdogan, T.; Zhong, H.; S ein, M.N.; Kim, I.Y.; Dipaola, R.S.;
Be ino, J.R.; Sabaawy, H.E. En ichmen o human p os a e cance cells wi h umo ini ia ing p ope ies in
mouse and zeb a ish xenog a s by di e en ial adhesion. P os a e 2014,74, 187–200. [C ossRe ] [PubMed]
117.
Zhang, B.; Shimada, Y.; Ku oyanagi, J.; Umemo o, N.; Nishimu a, Y.; Tanaka, T. Quan i a i e
pheno yping-based
in i o
chemical sc eening in a zeb a ish model o leukemia s em cell
xeno ansplan a ion. PLoS ONE 2014,9, 1–9. [C ossRe ] [PubMed]
118.
B own, H.K.; Schia one, K.; Tazzyman, S.; Heymann, D.; Chico, T.J.A. Zeb a ish xenog a models o cance
and me as asis o d ug disco e y. Expe Opin. D ug Disco . 2017,12, 379–389. [C ossRe ] [PubMed]
119.
Mo , R.L.; Jackson, I.J.; Pa on, E.E. The melanocy e lineage in de elopmen and disease. De elopmen
2015
,
142, 1387. [C ossRe ] [PubMed]
120.
Xie, X.; Ross, J.L.; Cowell, J.K.; Teng, Y. Fu u e Medicinal Chemis y. Fu u e Med. Chem.
2015
,7, 1395–1405.
[C ossRe ] [PubMed]
121.
Kim, K.-T.; Tanguay, R.L. In eg a ing zeb a ish oxicology and nanoscience o sa e p oduc de elopmen .
G een Chem. 2013,15, 872. [C ossRe ] [PubMed]
Genes 2017,8, 349 17 o 20
122.
Ha pe , B.; Thomas, D.; Chikkagouda , S.; Bake , N.; Tang, K.; He edia-Langne , A.; Lins, R.; Ha pe , S.
Compa a i e haza d analysis and oxicological modeling o di e se nanoma e ials using he emb yonic
zeb a ish (EZ) me ic o oxici y. J. Nanopa icle Res. 2015,17, 1–12. [C ossRe ] [PubMed]
123.
Jeong, J.; Cho, H.J.; Choi, M.; Lee, W.S.; Chung, B.H.; Lee, J.S.
In i o
oxici y assessmen o angiogenesis and
he li e dis ibu ion o nano-g aphene oxide and i s PEGyla ed de i a i es using he de eloping zeb a ish
emb yo. Ca bon N. Y. 2015,93, 431–440. [C ossRe ]
124.
Yang, T.; Ma in, P.; Foga y, B.; B own, A.; Schu man, K.; Phipps, R.; Yin, V.P.; Lockman, P.; Bai, S. Exosome
Deli e ed An icance D ugs Ac oss he Blood-B ain Ba ie o B ain Cance The apy in Danio Re io.
Pha m. Res. 2015,32, 2003–2014. [C ossRe ] [PubMed]
125.
Li, Y.; Miao, X.; Chen, T.; Yi, X.; Wang, R.; Zhao, H.; Lee, S.M.Y.; Wang, X.; Zheng, Y. Zeb a ish as a isual
and dynamic model o s udy he anspo o nanosized d ug deli e y sys ems ac oss he biological ba ie s.
Coll. Su . B Bioin e aces 2017,156, 227–235. [C ossRe ] [PubMed]
126.
Siebe , S.; G ossen, P.; De ampel, P.; Sieg ied, S.; Wi zigmann, D.; Huwyle , J. Zeb a ish as an ea ly
s age sc eening ool o s udy he sys emic ci cula ion o nanopa icula e d ug deli e y sys ems
in i o
.
J. Con ol. Release 2017,264, 180–191. [C ossRe ] [PubMed]
127.
E ensen, L.; Johansen, P.L.; Kos e , G.; Zhu, K.; He indal, L.; Spe h, M.; Fena oli, F.; Hildahl, J.; Baghe i am, S.;
Tulo a, C.; P asmickai e, L.; Mælandsmo, G.M.; Snaa -Jagalska, E.; G i i hs, G. Zeb a ish as a model sys em
o cha ac e iza ion o nanopa icles agains cance . Nanoscale 2016,8, 862–877. [C ossRe ] [PubMed]
128.
Wehmas, L.C.; Tanguay, R.L.; Punnoose, A.; G eenwood, J.A. De eloping a No el Emb yo–La al Zeb a ish
Xenog a Assay o P io i ize Human Glioblas oma The apeu ics. Zeb a ish
2016
,13, 317–329. [C ossRe ]
[PubMed]
129.
Yang, J.; Shimada, Y.; Ols hoo n, R.C.L.; Snaa -Jagalska, B.E.; Spaink, H.P.; K os, A. Applica ion o Coiled
Coil Pep ides in Liposomal An icance D ug Deli e y Using a Zeb a ish Xenog a Model. ACS Nano
2016
,
10, 7428–7435. [C ossRe ] [PubMed]
130.
Xu, B.; Jin, Q.; Zeng, J.; Yu, T.; Chen, Y.; Li, S.; Gong, D.; He, L.; Tan, X.; Yang, L.; He, G.; Wu, J.; Song, X.
Combined Tumo - and Neo ascula -“Dual Ta ge ing” Gene/Chemo-The apy Supp esses Tumo G ow h
and Angiogenesis. ACS Appl. Ma e . In e aces 2016,8, 25753–25769. [C ossRe ] [PubMed]
131.
Ald ian, G.; Vaissiè e, A.; Kona e, K.; Seisel, Q.; Vi ès, E.; Fe nandez, F.; Viguie , V.; Gene ois, C.; Couillaud, F.;
Démèné, H.; e al. PEGyla ion a e in luences pep ide-based nanopa icles media ed siRNA deli e y
in i o
and in i o. J. Con ol. Release 2017,256, 79–91. [C ossRe ] [PubMed]
132.
Co dei o, M.; Ca alho, L.; Sil a, J.; Saúde, L.; Fe nandes, A.; Bap is a, P. Gold Nanobeacons o T acking
Gene Silencing in Zeb a ish. Nanoma e ials 2017,7, 10. [C ossRe ] [PubMed]
133.
Zou, D.; Wang, W.; Lei, D.; Yin, Y.; Ren, P.; Chen, J.; Yin, T.; Wang, B.; Wang, G.; Wang, Y.
Pene a ion o blood-b ain ba ie and an i umo ac i i y and ne e epai in glioma by doxo ubicin-loaded
monosialoganglioside micelles sys em. In . J. Nanomed. 2017,12, 4879–4889. [C ossRe ] [PubMed]
134.
Collins, A.R.; Annangi, B.; Rubio, L.; Ma cos, R.; Do n, M.; Me ke , C.; Es ela-Lopis, I.; Cimpan, M.R.;
Ib ahim, M.; Cimpan, E.; e al. High h oughpu oxici y sc eening and in acellula de ec ion o
nanoma e ials. Wiley In e discip. Re . Nanomed. Nanobio echnol. 2017,9. [C ossRe ] [PubMed]
135.
B and, W.; Noo lande , C.W.; Giannakou, C.; Pa k, M.V.D.Z.; Vandeb iel, R.J.; Bosselae s, I.E.M.
Nanomedicinal p oduc s: A su ey on speci ic oxici y and side e ec s. In . J. Nanomed.
2017
,12, 6107–6129.
[C ossRe ] [PubMed]
136.
Giannakou, C.; Pa k, M.V.D.Z.; De Jong, W.H.; Van Lo e en, H.; Vandeb iel, R.J.; Gee sma, R.E.
A compa ison o immuno oxic e ec s o nanomedicinal p oduc s wi h egula o y immuno oxici y es ing
equi emen s. In . J. Nanomed. 2016,11, 2935–2952. [C ossRe ] [PubMed]
137.
Omidi, Y.; Hollins, A.J.; D ay on, R.M.; Akh a , S. Polyp opylenimine dend ime -induced gene exp ession
changes: The e ec o complexa ion wi h DNA, dend ime gene a ion and cell ype. J. D ug Ta ge .
2005
,13,
431–443. [C ossRe ] [PubMed]
138.
Poma, A.; Di Gio gio, M.L. Toxicogenomics o imp o e comp ehension o he mechanisms unde lying
esponses o
in i o
and
in i o
sys ems o nanoma e ials: A e iew. Cu . Genom.
2008
,9, 571–585.
[C ossRe ] [PubMed]
139.
Shah, V.; Ta a ula, O.; Ga buzenko, O.B.; Pa il, M.L.; Sa la, R.; Zhang, M.; Minko, T. Geno oxici y o di e en
nanoca ie s: Possible modi ica ions o he deli e y o nucleic acids. Cu . D ug Disco . Technol.
2013
,10,
8–15. [C ossRe ] [PubMed]
Genes 2017,8, 349 18 o 20
140.
Ancho doquy, T.J.; Ba enholz, Y.; Bo aschi, D.; Cho ny, M.; Decuzzi, P.; Dob o olskaia, M.A.;
Fa hang azi, Z.S.; Fa ell, D.; Gabizon, A.; Ghandeha i, H.; e al. Mechanisms and Ba ie s in Cance
Nanomedicine: Add essing Challenges, Looking o Solu ions. ACS Nano
2017
,11, 12–18. [C ossRe ]
[PubMed]
141.
Guideline, T.T.; Guideline, O. OECD Guidelines o he Tes ing o Chemicals; Oecd/Ocde 220; O ganisa ion o
Economic Co-ope a ion and De elopmen : Pa is, F ance, 2004; pp. 1–22. [C ossRe ]
142.
O ganisa ion o Economic Co-ope a ion and De elopmen (OECD). Tes No. 236: Fish Emb yo Acu e Toxici y
(FET) Tes ; OECD Guidelines o he Tes ing o Chemicals, Sec ion 2; OECD: Pa is, F ance, 2013; pp. 1–22.
[C ossRe ]
143.
Fen , K.; Weisb od, C.J.; Wi h-Helle , A.; Pieles, U. Assessmen o up ake and oxici y o luo escen silica
nanopa icles in zeb a ish (Danio e io) ea ly li e s ages. Aqua . Toxicol.
2010
,100, 218–228. [C ossRe ]
[PubMed]
144.
Ga cia, G.R.; Noyes, P.D.; Tanguay, R.L. Ad ancemen s in zeb a ish applica ions o 21s cen u y oxicology.
Pha macol. The . 2016,161, 11–21. [C ossRe ] [PubMed]
145.
Ong, K.J.; Zhao, X.; This le, M.E.; Macco mack, T.J.; Cla k, R.J.; Ma, G.; Ma inez-Rubi, Y.; Sima d, B.;
Loo, J.S.C.; Veino , J.G.C.; Goss, G.G. Mechanis ic insigh s in o he e ec o nanopa icles on zeb a ish ha ch.
Nano oxicology 2014,8, 295–304. [C ossRe ] [PubMed]
146.
Suka di, H.; Ung, C.Y.; Gong, Z.; Lam, S.H. Inco po a ing zeb a ish omics in o chemical biology and
oxicology. Zeb a ish 2010,7, 41–52. [C ossRe ] [PubMed]
147.
Hussainzada, N.; Lewis, J.A.; Bae , C.E.; Ippoli o, D.L.; Jackson, D.A.; S allings, J.D. Whole adul o ganism
ansc ip ional p o iling o acu e me al exposu es in male zeb a ish. BMC Pha macol. Toxicol.
2014
,15, 15.
[C ossRe ] [PubMed]
148.
Ge oy, B.; Ladha , C.; Cambie , S.; T egue -Delapie e, M.; B è hes, D.; Bou dineaud, J.-P. Impac o die a y
gold nanopa icles in zeb a ish a e y low con amina ion p essu e: The ole o size, concen a ion and
exposu e ime. Nano oxicology 2012,6, 144–160. [C ossRe ] [PubMed]
149.
Segu a-Aguila , J.; Kos zewa, R.M. Neu o oxins and neu o oxici y mechanisms. An o e iew. Neu o ox. Res.
2006,10, 263–287. [C ossRe ] [PubMed]
150.
MacPhail, R.C.; Hun e , D.L.; I ons, T.D.; Padilla, S. Locomo ion and Beha io al Toxici y in La al Zeb a ish:
Backg ound, Me hods, and Da a. In Zeb a ish; John Wiley & Sons, Inc.: Hoboken, NJ, USA, 2011; pp. 151–164,
ISBN 9781118102138.
151.
T uong, L.; Saili, K.S.; Mille , J.M.; Hu chison, J.E.; Tanguay, R.L. Pe sis en adul zeb a ish beha io al
de ici s esul s om acu e emb yonic exposu e o gold nanopa icles. Comp. Biochem. Physiol. Pa C
Toxicol. Pha macol. 2012,155, 269–274. [C ossRe ] [PubMed]
152.
Usenko, C.Y.; Ha pe , S.L.; Tanguay, R.L.
In i o
e alua ion o ca bon ulle ene oxici y using emb yonic
zeb a ish. Ca bon N. Y. 2007,45, 1891–1898. [C ossRe ] [PubMed]
153.
Vibe, C.B.; Fena oli, F.; Pi es, D.; Wilson, S.R.; Bogoe a, V.; Kallu u, R.; Spe h, M.; Anes, E.; G i i hs, G.;
Hildahl, J. Thio idazine in PLGA nanopa icles educes oxici y and imp o es i ampicin he apy agains
mycobac e ial in ec ion in zeb a ish. Nano oxicology 2016,10, 680–688. [C ossRe ] [PubMed]
154.
Zhuang, S.; Zhang, Z.; Zhang, W.; Bao, L.; Xu, C.; Zhang, H. Enan ioselec i e de elopmen al oxici y and
immuno oxici y o py aclo os owa d zeb a ish (Danio e io). Aqua . Toxicol.
2015
,159, 119–126. [C ossRe ]
[PubMed]
155.
Xu, H.; Dong, X.; Zhang, Z.; Yang, M.; Wu, X.; Liu, H.; Lao, Q.; Li, C. Assessmen o immuno oxici y
o dibu yl ph hala e using li e zeb a ish emb yos. Fish Shell ish Immunol.
2015
,45, 286–292. [C ossRe ]
[PubMed]
156.
Da oczi, B.; Ka i, G.; McAlee , M.F.; Wol , J.C.; Rodeck, U.; Dicke , A.P.
In i o
Radiop o ec ion by he
Fulle ene Nanopa icle DF-1 as Assessed in a Zeb a ish Model. Clin. Cance Res.
2006
,12, 7086–7091.
[C ossRe ] [PubMed]
157.
Sheng, L.; Wang, L.; Su, M.; Zhao, X.; Hu, R.; Yu, X.; Hong, J.; Liu, D.; Xu, B.; Zhu, Y.; Wang, H.; Hong, F.
Mechanism o TiO
2
nanopa icle-induced neu o oxici y in zeb a ish (D anio e io). En i on. Toxicol.
2016
,31,
163–175. [C ossRe ] [PubMed]
158.
Wang, J.; Zhu, X.; Zhang, X.; Zhao, Z.; Liu, H.; Geo ge, R.; Wilson-Rawls, J.; Chang, Y.; Chen, Y. Dis up ion o
zeb a ish (Danio e io) ep oduc ion upon ch onic exposu e o TiO2 nanopa icles. Chemosphe e
2011
,83,
461–467. [C ossRe ] [PubMed]
Genes 2017,8, 349 19 o 20
159.
Jang, G.H.; Lee, K.Y.; Choi, J.; Kim, S.H.; Lee, K.H. Mul i ace ed oxici y assessmen o ca alys composi es in
ansgenic zeb a ish emb yos. En i on. Pollu . 2016,216, 755–763. [C ossRe ] [PubMed]
160.
Lin, S.; Zhao, Y.; Xia, T.; Meng, H.; Ji, Z.; Liu, R.; Geo ge, S.; Xiong, S.; Wang, X.; Zhang, H.; e al. High
con en sc eening in zeb a ish speeds up haza d anking o ansi ion me al oxide nanopa icles. ACS Nano
2011,5, 7284–7295. [C ossRe ] [PubMed]
161.
Chang, J.; Ichiha a, G.; Shimada, Y.; Tada-Oikawa, S.; Ku oyanagi, J.; Zhang, B.; Suzuki, Y.; Sehsah, R.;
Ka o, M.; Tanaka, T.; e al. Coppe Oxide Nanopa icles Reduce Vasculogenesis in T ansgenic Zeb a ish
Th ough Down-Regula ion o Vascula Endo helial G ow h Fac o Exp ession and Induc ion o Apop osis.
J. Nanosci. Nano echnol. 2015,15, 2140–2147. [C ossRe ] [PubMed]
162.
Ba -Ilan, O.; Louis, K.M.; Yang, S.P.; Pede sen, J.A.; Hame s, R.J.; Pe e son, R.E.; Heideman, W. Ti anium
dioxide nanopa icles p oduce pho o oxici y in he de eloping zeb a ish. Nano oxicology
2012
,6, 670–679.
[C ossRe ] [PubMed]
163.
Zhang, W.; Lin, K.; Sun, X.; Dong, Q.; Huang, C.; Wang, H.; Guo, M.; Cui, X. Toxicological e ec o MPA–CdSe
QDs exposu e on zeb a ish emb yo and la ae. Chemosphe e 2012,89, 52–59. [C ossRe ] [PubMed]
164.
Powell, D.R.; Hu enloche , A. Neu ophils in he Tumo Mic oen i onmen . T ends Immunol.
2016
,37, 41–52.
[C ossRe ] [PubMed]
165.
Chambe s, S.E.J.; O’Neill, C.L.; O’Dohe y, T.M.; Medina, R.J.; S i , A.W. The ole o immune- ela ed myeloid
cells in angiogenesis. Immunobiology 2013,218, 1370–1375. [C ossRe ] [PubMed]
166.
Renshaw, S.A.; Loynes, C.A.; T ushell, D.M.I.; Elwo hy, S.; Ingham, P.W.; Why e, M.K.B. A ansgenic
zeb a ish model o neu ophilic in lamma ion. Blood 2006,108, 3976–3978. [C ossRe ] [PubMed]
167.
Duan, J.; Hu, H.; Li, Q.; Jiang, L.; Zou, Y.; Wang, Y.; Sun, Z. Combined oxici y o silica nanopa icles and
me hylme cu y on ca dio ascula sys em in zeb a ish (Danio e io) emb yos. En i on. Toxicol. Pha macol.
2016,44, 120–127. [C ossRe ] [PubMed]
168.
Bousheh i, M.A.S.; Lamp ech , A. Nanopa icles as d ug ca ie s: Cu en issues wi h
in i o
es ing.
Nanomedicine 2015,10, 3213–3230. [C ossRe ] [PubMed]
169.
Nguyen, V.H.; Lee, B.J. P o ein co ona: A new app oach o nanomedicine design. In . J. Nanomed.
2017
,12,
3137–3151. [C ossRe ] [PubMed]
170.
Jain, P.; Pawa , R.S.; Pandey, R.S.; Madan, J.; Pawa , S.; Lakshmi, P.K.; Sudheesh, M.S. In- i o in- i o
co ela ion (IVIVC) in nanomedicine: Is p o ein co ona he missing link? Bio echnol. Ad .
2017
,35, 889–904.
[C ossRe ] [PubMed]
171.
Ha pe , S.; Maddux, B.L.S.; Hu chison, J.E.; Usenko, C.; Tanguay, R. Biodis ibu ion and Toxici y o
Nanoma e ials In Vi o: E ec s o Composi ion, Size, Su ace Func ionaliza ion and Rou e o Exposu e.
Nano ech 2007,2, 666–669.
172.
Wilhelm, S.; Ta a es, A.J.; Dai, Q.; Oh a, S.; Aude , J.; D o ak, H.F.; Chan, W.C.W. Analysis o nanopa icle
deli e y o umou s. Na . Re . Ma e . 2016,1. [C ossRe ]
173. To ice, M. Does Nanomedicine Ha e a Deli e y P oblem? ACS Publ. 2016. [C ossRe ] [PubMed]
174.
Mun imadugu, E.; Kommineni, N.; Khan, W. Explo ing he Po en ial o Nano he apeu ics in Ta ge ing
Tumo Mic oen i onmen o Cance The apy. Pha macol. Res. 2017, 1–14. [C ossRe ] [PubMed]
175.
S ylianopoulos, T.; Jain, R.K. Design conside a ions o nano he apeu ics in oncology. Nanomedicine
2015
,11,
1893–1907. [C ossRe ] [PubMed]
176.
Kunin y, P.R.; Schni e , J.; S o m, G.; P akash, J. Mic oRNA Ta ge ing o Modula e Tumo Mic oen i onmen .
F on . Oncol. 2016,6, 1–8. [C ossRe ] [PubMed]
177.
Ki chbe ge , S.; S u zel, C.; Pascoal, S.; Dis el, M. Quo na as, Danio?—Recen P og ess in Modeling Cance in
Zeb a ish. F on . Oncol. 2017,7. [C ossRe ] [PubMed]
178.
Wang, J.; Cao, Z.; Zhang, X.M.; Nakamu a, M.; Sun, M.; Ha man, J.; Ha is, R.A.; Sun, Y.; Cao, Y. No el
mechanism o mac ophage-media ed me as asis e ealed in a zeb a ish model o umo de elopmen .
Cance Res. 2015,75, 306–315. [C ossRe ] [PubMed]
179.
Rouhi, P.; Jensen, L.D.; Cao, Z.; Hosaka, K.; Länne, T.; Wahlbe g, E.; S e ensen, J.F.; Cao, Y. Hypoxia-induced
me as asis model in emb yonic zeb a ish. Na . P o oc. 2010,5, 1911–1918. [C ossRe ] [PubMed]
180.
S ole o , K.; Mon el, V.; Les e , R.D.; Gonias, S.L.; Klemke, R. High- esolu ion imaging o he dynamic umo
cell ascula in e ace in anspa en zeb a ish. P oc. Na l. Acad. Sci. USA
2007
,104, 17406–17411. [C ossRe ]
[PubMed]
Genes 2017,8, 349 20 o 20
181.
Van de En , W.; Bu ello, C.; de Lange, M.J.; an de Velden, P.A.; Jochemsen, A.G.; Jage , M.J.;
Snaa -Jagalska, B.E. Emb yonic Zeb a ish: Di e en Pheno ypes a e Injec ion o Human U eal Melanoma
Cells. Ocul. Oncol. Pa hol. 2015,1, 170–181. [C ossRe ] [PubMed]
182.
Zhang, B.; Shimada, Y.; Ku oyanagi, J.; Nishimu a, Y.; Umemo o, N.; Nomo o, T.; Shin ou, T.; Miyazaki, T.;
Tanaka, T. Zeb a ish xeno ansplan a ion model o cance s em-like cell s udy and high- h oughpu
sc eening o inhibi o s. Tumo Biol. 2014,35, 11861–11869. [C ossRe ] [PubMed]
183.
Eguia a, A.; Holgado, O.; Beloqui, I.; Abalde, L.; Sanchez, Y.; Callol, C.; Ma in, A.G. Xenog a s in zeb a ish
emb yos as a apid unc ional assay o b eas cance s em-like cell iden i ica ion. Cell Cycle
2011
,10,
3751–3757. [C ossRe ] [PubMed]
184.
Zhao, Y.; Huang, X.; Ding, T.W.; Gong, Z. Enhanced angiogenesis, hypoxia and neu ophil ec ui men
du ing Myc-induced li e umo igenesis in zeb a ish. Sci. Rep. 2016,6, 31952. [C ossRe ] [PubMed]
185.
Wagne , D.S.; Delk, N.A.; Lukiano a-hleb, E.Y.; Ha ne , J.H.; Lapo ko, D.O. The
in i o
pe o mance o
plasmonic nanobubbles as cell he anos ic agen s in zeb a ish hos ing p os a e cance xenog a s. Bioma e ials
2010,31, 7567–7574. [C ossRe ] [PubMed]
186.
Jain, A.S.; Makhija, D.T.; Goel, P.N.; Shah, S.M.; Nikam, Y.; Gude, R.P.; Jag ap, A.G.; Naga senke , M.S.
Doce axel in ca ionic lipid nanocapsules o enhanced
in i o
ac i i y. Pha m. De . Technol.
2016
,21, 76–85.
[C ossRe ] [PubMed]
187.
Gong, C.; Deng, S.; Wu, Q.; Xiang, M.; Wei, X.; Li, L.; Gao, X.; Wang, B.; Sun, L.; Chen, Y.; e al. Imp o ing
an iangiogenesis and an i- umo ac i i y o cu cumin by biodeg adable polyme ic micelles. Bioma e ials
2013,34, 1413–1432. [C ossRe ] [PubMed]
188.
Nabe , H.P.H.; D absch, Y.; Snaa -Jagalska, B.E.; en Dijke, P.; an Laa , T. Snail and Slug, key egula o s o
TGF-
β
-induced EMT, a e su icien o he induc ion o single-cell in asion. Biochem. Biophys. Res. Commun.
2013,435, 58–63. [C ossRe ] [PubMed]
189.
Yang, X.J.; Cui, W.; Gu, A.; Xu, C.; Yu, S.C.; Li, T.T.; Cui, Y.H.; Zhang, X.; Bian, X.W. A No el Zeb a ish
Xeno ansplan a ion Model o S udy o Glioma S em Cell In asion. PLoS ONE
2013
,8, 1–9. [C ossRe ]
[PubMed]
190.
He, S.; Lame s, G.E.M.; Beenakke , J.W.M.; Cui, C.; Gho a, V.P.S.; Danen, E.H.J.; Meije , A.H.; Spaink, H.P.;
Snaa -Jagalska, B.E. Neu ophil-media ed expe imen al me as asis is enhanced by VEGFR inhibi ion in a
zeb a ish xenog a model. J. Pa hol. 2012,227, 431–445. [C ossRe ] [PubMed]
191.
Tobia, C.; Ga iano, G.; De Sena, G.; P es a, M. Zeb a ish emb yo as a ool o s udy umo /endo helial cell
c oss- alk. Biochim. Biophys. Ac a 2013,1832, 1371–1377. [C ossRe ] [PubMed]
192.
Duan, J.; Hu, H.; Feng, L.; Yang, X.; Sun, Z. Silica nanopa icles inhibi mac ophage ac i i y and angiogenesis
ia VEGFR2-media ed MAPK signaling pa hway in zeb a ish emb yos. Chemosphe e
2017
,183, 483–490.
[C ossRe ] [PubMed]
193.
Van Ham, T.J.; Mapes, J.; Kokel, D.; Pe e son, R.T. Li e imaging o apop o ic cells in zeb a ish. FASEB J.
2010
,
24, 4336–4342. [C ossRe ] [PubMed]
194.
Chak abo y, C.; Sha ma, A.R.; Sha ma, G.; Lee, S.-S. Zeb a ish: A comple e animal model o enume a e he
nanopa icle oxici y. J. Nanobio echnol. 2016,14, 65. [C ossRe ] [PubMed]
195.
Blechinge , S.R.; E ans, T.G.; Tang, P.T.; Kuwada, J.Y.; Wa en, J.T., J .; K one, P.H. The hea -inducible
zeb a ish hsp70 gene is exp essed du ing no mal lens de elopmen unde non-s ess condi ions. Mech. De .
2002,112, 213–215. [C ossRe ]
©
2017 by he au ho s. Licensee MDPI, Basel, Swi ze land. This a icle is an open access
a icle dis ibu ed unde he e ms and condi ions o he C ea i e Commons A ibu ion
(CC BY) license (h p://c ea i ecommons.o g/licenses/by/4.0/).
