Models o s udy basic and applied aspec s o lysosomal s o age diso de s
Ángel Gaudioso
a
, Te esa P. Sil a
b
, Ma ía Dolo es Ledesma
a,
⇑
a
Cen o Biología Molecula Se e o Ochoa (CSIC-UAM), Mad id, Spain
b
Ins i u o de Medicina Molecula João Lobo An unes, Faculdade de Medicina, Uni e sidade de Lisboa, Po ugal
a icle in o
A icle his o y:
Recei ed 29 Oc obe 2021
Re ised 5 Augus 2022
Accep ed 4 Sep embe 2022
A ailable online 16 Sep embe 2022
Keywo ds:
Lysosomes
S o age diso de s
Omics
Op ogene ics
iPSCs
O ganoids
BBB
Animal models
abs ac
The lack o a ailable ea men s and a al ou come in mos lysosomal s o age diso de s (LSDs) ha e
spu ed esea ch on pa hological mechanisms and no el he apies in ecen yea s. In his e o , expe i-
men al me hodology in cellula and animal models ha e been de eloped, wi h aims o add ess majo
challenges in many LSDs such as pa ien - o-pa ien a iabili y and b ain condi ion. These echniques
and models ha e ad anced knowledge no only o LSDs bu also o o he lysosomal diso de s and ha e
p o ided undamen al insigh s in o he biological oles o lysosomes. They can also se e o assess he
e icacy o classical he apies and mode n d ug deli e y sys ems. He e, we summa ize he echniques
and models used in LSD esea ch, which include bo h es ablished and ecen ly de eloped in i o me h-
ods, wi h gene al u ili y o speci ically add essing lysosomal ea u es. We also e iew animal models o
LSDs oge he wi h cu ing-edge echnology ha may educe he need o animals in he s udy o hese
de as a ing diseases.
Ó2022 The Au ho (s). Published by Else ie B.V. This is an open access a icle unde he CC BY license
(h p://c ea i ecommons.o g/licenses/by/4.0/).
Con en s
1. In oduc ion . . . ........................................................................................................ 2
2. In i o echniques in LSD esea ch . . . ..................................................................................... 2
2.1. ‘‘Omics” . . . . . . . . . . . . . . ........................................................................................... 2
2.2. Fluo escence-based echniques . . . . . . . . . . . ........................................................................... 5
2.3. Elec on mic oscopy . . . . ........................................................................................... 7
2.4. Op ogene ics . . . . . . . . . . ........................................................................................... 7
2.5. Liposome-based echniques . . . . . . . . . . . . . . ........................................................................... 8
3. Cellula models in LSD esea ch . . . . . . ..................................................................................... 8
3.1. P ima y cell cul u es . . . . ........................................................................................... 8
3.2. Immo alized cell lines . . ........................................................................................... 8
3.3. Induced plu ipo en s em cells (iPSCs) . . . . . ........................................................................... 9
3.4. Mul ilineage cellula models: O ganoids . . . . .......................................................................... 10
3.5. BBB models . . . . . . . . . . . .......................................................................................... 10
4. Animal models in LSD esea ch. . . . . . . .................................................................................... 11
4.1. Na u ally occu ing animal models . . . . . . . . .......................................................................... 11
4.2. D osophila melanogas e .......................................................................................... 11
4.3. Caeno habdi is elegans . . .......................................................................................... 11
h ps://doi.o g/10.1016/j.add .2022.114532
0169-409X/Ó2022 The Au ho (s). Published by Else ie B.V.
This is an open access a icle unde he CC BY license (h p://c ea i ecommons.o g/licenses/by/4.0/).
Abb e ia ions: AAV9, Adenoassocia ed Vi al Vec o s se o ype 9; ASM, Acid Sphingomyelinase; ASMko, Acid Sphingomyelinase Knock-ou ; BBB, Blodd-B ain Ba ie ;
BODIPY, Bo on dipy ome hene; CNS, Cen al Ne ous Sys em; CRISPR, Clus e ed Regula ly In e spaced Sho Palind omic Repea s; EM, Elec on Mic oscopy; ER, Endoplasmic
Re iculum; GAL4/UAS, GAL4/Ups eam Ac i a ing Sequence; GFP, G een Fluo escen P o ein; hiPSCs, human induced Plu ipo en S em Cells; iPSCs, induced Plu ipo en S em
Cells; LAMP1/2, Lysosomal Associa ed Memb ane P o ein 1/2; LPC, Lysophospha idylcholine; LPS, Lysophospha idylse ine; LSD, Lysosomal S o age Diso de ; mTORC1,
mammalian Ta ge O Rapamicine Complex 1; NBD, Ni obenzoxadiazole; OMIM, Online Mendelian Inhe i ance in Man; PC, Phospha idylcholine; PE, Phos-
pha idyle hanolamine; PS, Phospha idylse ine; PC-O, Plasmenylcholine; PE-O, Plasmenyle hanolamine.
⇑
Co esponding au ho a : Cen o Biologia Molecula Se e o Ochoa, Nicolas Cab e a 1, 28049 Mad id, Spain.
E-mail add ess: [email p o ec ed] (M. Dolo es Ledesma).
Ad anced D ug Deli e y Re iews 190 (2022) 114532
Con en s lis s a ailable a ScienceDi ec
Ad anced D ug Deli e y Re iews
jou nal homepage: www.else ie .com/loca e/ad
4.4. Zeb a ish . . . . . . . . . . . . . .......................................................................................... 12
4.5. Mu ine models . . . . . . . . .......................................................................................... 13
4.6. Non-human p ima es . . . .......................................................................................... 13
5. Conclusion . . . . ....................................................................................................... 13
Decla a ion o Compe ing In e es . . . . .................................................................................... 13
Acknowledgemen s . . . . . . ............................................................................................. 14
Funding . . . . . . . . ....................................................................................................... 14
Re e ences . . . . ....................................................................................................... 14
1. In oduc ion
Due o hei key ole in he b eakdown o cellula componen s
[1], and o many yea s since hei disco e y in 1950 [2], lysosomes
we e conside ed me ely ‘‘cellula ash cans”. Nowadays, a
ple ho a o unc ions beyond he deg ada ion o in a- and ex a-
cellula molecules and o ganelles [3] ha e been assigned o lyso-
somes, including nu ien sensing [4], he quali y con ol sys em
in ol ed in he ageing p ocess [5] and pa icipa ion in inna e and
adap i e immuni y [6,7] (Fig. 1).
Aside om hei in ol emen in mul iple cellula p ocesses,
lysosomes make physical connec ions wi h se e al in acellula
o ganelles such as mi ochond ia [8], he endoplasmic e iculum
[9] o plasma memb ane [10] by memb ane e he ing and usion.
These con ac si es a e c ucial o he co ec unc ion o no only
lysosomes, bu also he subcellula o ganelles ha use wi h hem
[11,12]. P ope lysosomal ac i i y is i al o adequa e cell, issue
and o gan unc ions, and lysosomal impai men plays a ele an
pa hological ole in many di e en diseases including cance [13]
and neu odegene a i e [14–16] o au oimmune diseases [17].
Among he lysosome- ela ed diseases a e he lysosomal s o age
diso de s (LSDs), whose ae iology is di ec ly linked o lysosomal
dys unc ion. Mos LSDs a e caused by mu a ions in genes ha
encode lysosomal enzymes, esul ing in mo e han 70 di e en
a e diseases cha ac e ised by he lysosomal accumula ion o
molecules, in many cases he e y subs a es o he de ec i e
enzymes [18].
One way o classi y LSDs is acco ding o he ype o molecule
ha accumula es in he lysosomes. This classi ica ion scheme
(Table 1), which will be used o o ganize his e iew, includes h ee
majo g oups: sphingolipidoses, mucopolysaccha idoses and
glycop o einoses/oligosaccha idoses [53] (Fig. 1). Sphingolipidoses
a e caused by de ec s in he lysosomal deg ada ion o sphin-
golipids, and he accumula ion o lipid in hese diseases causes
memb ane de ec s ha can a ec cell su i al, especially o neu-
ons. As a esul , neu odegene a ion along wi h isce al complica-
ions a e cha ac e is ic o many sphingolipidoses [54].
Mucopolysaccha idoses a e he consequence o de ec s in he
deg ada ion o glycosaminoglycans (complex amino-suga poly-
me s). These diseases p og ess wi h bo h pe iphe al (i.e. uppe ai -
way obs uc ion o ca diac al e and muscle disease) and
neu opa hic (i.e. hype ac i i y, impai ed de elopmen and loss o
cogni i e unc ion) ea u es [55]. The hi d g oup,
glycop o einoses/oligosaccha idoses esul om al e a ions in ca -
abolism o oligosaccha ide chains om glycolipids and glycop o-
eins, leading o he accumula ion o hese molecules. Thei
clinical mani es a ions a e simila o hose al eady men ioned o
he mucopolysaccha idoses [56].
Besides he LSDs di ec ly caused by he impai ed unc ion o
lysosomal me abolic enzymes, some LSDs a e he consequence o
al e a ions in p o eins ha con ol a icking o molecules h ough
he endolysosomal compa men [57] o in lysosomal memb ane
p o eins. This g oup o diso de s a e also cha ac e ized by s o age
inside lysosomes. Rep esen a i e examples a e Niemann-Pick ype
C (OMIM 257220; 607625), which esul s om mu a ions in he
choles e ol anspo p o eins NPC1 o NPC2 [58,59] o Danon dis-
ease (OMIM 300257), which is caused by mu a ions in he gene
encoding he in eg al lysosomal memb ane p o ein LAMP2 [60].
Iden i ica ion o he gene ic cause in many LSDs has no been
accompanied by a deep knowledge o he pa hological mecha-
nisms, much less by e icien ea men s. Among he impo an
ca ea s o cu en LSD he apies is he deli e y o he apeu ics
o hese diseases. D ug deli e y sys ems may a ge and a ic
o lysosomes a e en e ing cells. Howe e , accumula ion o he a-
peu ics in hese o ganelles p io o deg ada ion could cause dele e-
ious e ec s. In addi ion, d ug deli e y sys ems ha aim o dis u b
he endolysosomal compa men , so ha he con ained he apeu-
ics can escape in o he cy osol and each o he in acellula a -
ge s, may also a ec lysosomal unc ion. Pa icula ly challenging
is he deli e y o he apeu ics in o he b ain, which is necessa y
o ea he nume ous neu ological LSDs. Es ablished and newly
de eloped echniques and sui able cellula and animal models
a e equi ed no only o unde s and pa hological mechanisms in
LSDs bu also o assess lysosomal s a us and blood b ain ba ie
pene ance and o e alua e he apeu ic s a egies and d ug deli -
e y sys ems in hese diseases.
This e iew ocuses on he cu en s a e o such echniques and
models, conside s ad an ages and d awbacks (Fig. 2) gi ing exam-
ples o hei use in di e en ypes o LSDs in he pas , and discusses
in some ins ances he po en ial o ad ance he apeu ic d ug deli -
e y and de elopmen in he u u e.
2. In i o echniques in LSD esea ch
Nume ous in i o app oaches exis ha can be applied o s udy
lysosomal s uc u e and unc ion, as well as o diagnose and de e -
mine pa hological al e a ions and he apeu ic e icacy o LSDs. In
his sec ion we summa ise he es ablished app oaches ha a e
employed in he LSD con ex and echniques ha ha e mo e
ecen ly been de eloped o speci ically add ess lysosomes and
hei al e a ions.
2.1. ‘‘Omics”
‘‘Omics” is a g oup o echniques used o collec i ely cha ac e -
ize and quan i y pools o biological molecules, and hey can be
applied o a g ea a ie y o samples including cul u ed cells, ani-
mal models o pa ien -de i ed samples. Depending on he ype
o biological molecules ha a e cha ac e ized, ‘‘omics” can be
di ided in p o eomics, lipidomics, genomics, me abolomics, e c.
‘‘Omics” ools a e e y use ul in human clinical esea ch because
hey allow a gene al cha ac e iza ion o disease- ela ed al e a ions
in di e en issues and samples. In he case o gene ic diseases like
LSDs, genomic analysis can se e o de ec key mu a ions. Addi-
ionally, p o eomic and/o lipidomic analysis, which de ec al e -
a ions in he le els o p o eins o lipids, espec i ely, can gi e a
comp ehensi e iew o subs a es ha may accumula e in LSDs.
O he impo an applica ions o ‘‘omics” in LSD esea ch a e hose
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
2
Fig. 1. A. Schema ic o known lysosomal unc ions. B. LSD classi ica ion and di e en ypes o lysosomal anomalies ound in he LSD con ex indica ing he echniques ha
may help o analyse hem. EM: Elec on mic oscopy.
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
3
aimed a inding bioma ke s o diagnosis and assessmen o ea -
men e icacy.
Gi en he e sa ili y and insigh ha ‘‘omics” can p o ide, he e
a e nume ous s udies using hese echniques in he LSD con ex . In
he sphingolipidoses, p o eomic analysis o u ine, blood and issue
samples om pa ien s con ibu ed o he inding o new bioma k-
e s in Fab y disease (OMIM 301500) [61]. P o eomic p o iling o
lysosomes in Niemann-Pick ype C (OMIM 257220; 607625), in
which sphingolipids also accumula e, iden i ied mTORC1 signalling
as a a ge able pa hway o ea men [62]. Lipidomic analysis
enabling he simul aneous quan i ica ion o lysosphingolipids
allowed he sc eening o Fab y (OMIM 301500), Gauche (OMIM
230800), in an ile K abbe (OMIM 245200) and Niemann-Pick dis-
eases wi h high sensi i i y and speci ici y [63]. This analysis pe -
mi ed he disco e y o N-acyl-O-phosphocholinese ine, a class o
lipid ha had no been p e iously de ec ed in biological samples,
as a bioma ke in plasma om Niemann-Pick ype C (OMIM
257220; 607625) pa ien s [64]. Lipidomic p o iling o plasma and
u ine om Gauche disease (OMIM 230800) pa ien s was used o
iden i y lipid species, which le els we e signi ican ly al e ed and
could be modi ied by enzyme eplacemen he apy indica ing ha
hese species we e di ec ly o indi ec ly a ec ed by he he apy
[65]. Le els o plasma glucosylsphingosine a e being used as eli-
able ma ke o ollow he apeu ic esponse in Gauche (OMIM
230800) pa ien s [66]. An example o lipidomic analysis un eiling
di e ences in he b ain phospholipid con en in a mouse model o
he acid sphingomyelinase de iciency is shown in Fig. 3.
In mucopolysaccha idoses, b ain ansc ip omic analysis
assessed he e ec s o cen al ne ous sys em (CNS) gene he apy
in a mouse model o Hun e synd ome (OMIM 309900). Almos
80 % o he genes di e en ially exp essed, o which many associ-
a ed o in lamma ion and inna e immuni y, we e co ec ed a a sig-
ni ican ex en in hei ansc ip le els by adminis a ion o
adeno-associa ed i us ec o s encoding idu ona e-2-sul a ase
Table 1
Lis o ep esen a i e LSDs, classi ied acco ding o he na u e o he s o age molecule, indica ing he gene a ec ed, he encoded p o ein, he OMIM (Online Mendelian Inhe i ance
in Man) iden i ie and he p ima y accumula ing subs a e in each o hem. *Pompe disease is included in his g oup due o he accumula ion o polysaccha ides.
Disease Gene
a ec ed
OMIM Encoded p o ein P ima y accumula ing
molecule
Re
Sphingolipidoses
GM1 gangliosidosis GLB1 230,500 b-galac osidase GM1 ganglioside [19]
Sandho disease HEXB 268,800 Hexosaminidase A/B GM2 ganglioside [20]
Tay-Sachs disease HEXA 272,800 Hexosaminidase A GM2 ganglioside [21]
Fab y disease GLA 301,500
a
-galac osidase A Globo iaosylce amide [22]
Me ach oma ic
leukodys ophy
ARSA;
PSAP
250,100 A ylsul a ase A Sul a ides [23]
K abbe disease GALC 245,200 b-galac oce eb osidase Psychosine [24]
Niemann-Pick A/B
disease
SMPD1 257200;
607,616
Acid sphingomyelinase Sphingomyelin [25]
Fa be disease ASAH1 228,000 Acid ce amidase Ce amide [26]
Gauche disease GBA 230,800 Glucoce eb osidase Glucosylce amide [27]
Mucopolysaccha idoses
Hun e synd ome IDS 309,900 Idu ona e 2-sul a ase Hepa an and de ma an sul a e [28]
Hu le -Scheie synd ome IDUA 607,015
a
-l-idu onidase Hepa an and de ma an sul a e [29]
San ilippo synd ome A SGSH 252,900 Sul amidase Hepa an sul a e [30]
San ilippo synd ome C HGSNAT 252,930 Ace yl CoA
a
-glucosaminide
ace yl ans e ase
Hepa an sul a e [30]
San ilippo synd ome B NAGLU 252,920
a
-N-ace ylglucosaminidase Hepa an sul a e [30]
Sly disease GUSB 253,220 b-glucu onidase Chond oi in, hepa an and de ma an sul a e [31]
San ilippo synd ome D GNS 252,940 N-ace ylglucosamine-6-sul a ase Hepa an sul a e [30]
Mo quio A synd ome GALNS 253,000 N-ace ylglucosamine-6-sul a e sul a ase Ke a an and chond oi in-6-sul a e [32]
Mo quio B synd ome GLB1 253,010 b-galac osidase Ke a an and chond oi in-6-sul a e [33]
Ma o eaux-Lamy
synd ome
ARSB 253,200 A ylsul a ase B De ma an and chond oi in-4-sul a e [34]
Na owicz synd ome HYAL1 601,492 Hyalu onidase 1 Hyalu onan [35]
Glycop o einoses/Oligosaccha idoses
Sialidosis ype I NEU1 256,550 Neu aminidase Sialyloligosaccha ides [36]
Sialidosis ype II NEU1 256,550 Neu aminidase Sialyloligosaccha ides [37]
Galac osialidosis CTSA 256,540 Ca hepsin A Sialyloligosaccha ides [38]
Schindle disease NAGA 609,241
a
-N-ace ylgalac osaminidase Galac ose oligosaccha ides, galac omannans and
galac olipids
[39]
a
-mannosidosis MAN2B1 248,500
a
-
D
-mannosidase Mannose-con aining oligosaccha ides [40]
b-mannosidosis MANBA 248,510 b-mannosidase Di- and isaccha ides wi h b-linked mannose a he
non- educing end
[41]
Fucosidosis FUCA1 230,000
a
-
L
- ucosidase Fucosyla ed glycop o eins [42]
Aspa ylglycosaminu ia AGA 208,400 Glycosylaspa aginase Glycoaspa agines [43]
Pompe disease* GAA 232,300 Glucosidase Alpha, Acid Glycogen [44]
Non-enzyma ic LSDs
Danon disease LAMP2 300,257 Lysosome Associa ed Memb ane P o ein 2 Glycogen [45]
Niemann Pick ype C NPC1
NPC2
257220;
607,625
Niemann Pick ype C1
Niemann Pick ype C2
Choles e ol [46]
Salla disease SLC17A5 604,369 Sialin Sialic acid [47]
Cys inosis CTNS 219,800 Cys inosin F ee cys ine [48]
Mucolipidosis IV MCOLN1 252,650 Mucolipin-1 Phospholipids, gangliosides and mucopolysaccha ides [49]
Ba en disease CLN3 204,200 CLN3 Lipo uscin-like ma e ial [50]
Ce oid lipo uscinosis
ype 7
MFSD8 610,951 Majo acili a o supe amily domain-
con aining p o ein 8
Lipo uscin-like ma e ial [51]
Ce oid lipo uscinosis
ype 12
ATP13A2 606,693 Polyamine- anspo ing ATPase 13A2 Lipo uscin-like ma e ial [52]
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
4
[67]. Whole exome sequencing sc eened missense changes in he
SGSH gene in a g oup o adul onse San ilippo synd ome pa ien s
ha p esen ed e y mild symp oms aising awa eness o his unu-
sual pheno ype [68].
In glycop o einoses, a based-elec osp ay mass spec ome y
echnique o he de ec ion o ca bohyd a es was used o sc een
a complex mix u e o O-glycosyla ed sialyla ed amino acids om
u ine o a Schindle disease (OMIM 609241) pa ien [69] al hough
his me hod has no been widely adop ed o diagnosis. T ansc ip-
omic analysis o FUCA1 knock-down ke a inocy es un eiled he
al e ed exp ession o genes mainly ela ed o di e en ia ion and
immune esponse in ucosidosis (OMIM 230000) [70].
2.2. Fluo escence-based echniques
Fluo escence-based echniques ha e g ea ly acili a ed he
de ec ion o lysosomal damage and s o age by mic oscopy o spec-
opho ome y. Lysosomes can be isualized by immuno luo es-
cence echniques using an ibodies agains lysosomal memb ane
p o eins such as LAMP1 (Fig. 4A). Cell-pe meable luo escen
p obes a e a ailable ha a ge lysosomes. Fo example, he lyso-
acke ed dye s ains acidic compa men s in li e cells [71], and
has been used o moni o lysosomal pe meabiliza ion in LSD cellu-
la models [72,73] (Fig. 4B). SiR-lysosome is a luo ogenic and
highly speci ic p obe o lysosomes based on peps a in A, a na u al
Fig. 2. Illus a ion o cellula and animal models used in LSD esea ch indica ing hei main ad an ages (in g een) and d awbacks (in ed). Common ad an age o he cellula
models is he sui abili y o sc een d ugs and d ug deli e y sys ems and cha ac e ize pa hological molecula mechanisms. Common disad an ages a e he isola ion om he
physiological en i onmen and, excep o p ima y cul u es, he geno ypic/pheno ypic a ia ions ha may a ise du ing he cul u ing p ocess. Common ad an age o animal
models is he possibili y o s udying pa hology and he apy in he con ex o he whole o ganism. Howe e , species-speci ic di e ences in physiology, me abolism and
gene ics mus be aken in o accoun when ex apola ing esul s o he human condi ion. hiPSCs: Human-induced plu ipo en s em cells; BBB: Blood B ain Ba ie .
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
5

ca hepsin D-binding p oduc [74]. I s ains lysosomes in li e cells
wi hou he need o gene ic manipula ion o o e exp ession,
and has been used o moni o lysosome mo ili y by single-
pa icle acking [75]. Fluo escence-based echniques a e also
a ailable o moni o lysosomal unc ion. Ra iome ic assessmen
o lysosome pH using exogenous o gene ically encoded p obes
can be used no only o de e mine lysosomal pa hology in LSD
models bu also o assess he sa e y o d ug deli e y sys ems a -
ge ing he lysosome [76]. Newly de eloped exogenous p obes o
quan i y lysosome pH, such as CQ-Lyso, u ilize a single exci a ion
wa eleng h minimizing backg ound noise and au o luo escence
in e e ence. New gene ically encoded pH lysosome biosenso s
allow measu emen o lysosomal pH o e ex ended pe iods o ime
al hough i is necessa y o assess whe he hei o e exp ession has
any e ec on lysosomal size, posi ioning o ac i i y [76].
The e a e di e en ways o associa e luo escence wi h he
accumula ion o speci ic lipids in he LSD con ex , including luo-
escen analogues o na i e lipids (e.g. ni obenzoxadiazole [NBD]
o bo on-dipy ome hene [BODIPY-lipids]) [77,78]; au o luo es-
cen molecules such as he choles e ol-binding an ibio ic ilipin
[79], o pep ide sequences in oxins ha media e speci ic lipid
binding. Examples o he la e a e he ea hwo m oxin lysenin,
which binds sphingomyelin [80] (Fig. 5A,B); he D4 domain o pe -
ingolysin O, which binds choles e ol [81]; and he B subuni o
chole a oxin, which binds he ganglioside GM1 [82]. De ec ion
o hese oxin-de i ed pep ides by speci ic an ibodies, ans ec ion
o hei cDNAs used wi h luo escen p o eins o by adding o
hem luo escen ags, has helped cha ac e ize lysosomal s o age
in Niemann-Pick diseases [83,84]. They also allowed he disco e y
o lipid s o age beyond lysosomes, which a ec s o he cellula
compa men s wi h key ele ance o pa hology such as ha o
sphingomyelin a he plasma memb ane in Niemann-Pick ype A
(OMIM 257200) neu ons [80] (Fig. 5A).
The a o emen ioned p obes ha e been used o de e mine he
e icacy o po en ial he apeu ic s a egies. Reduc ion in ilipin
s aining con i med he use ulness o he choles e ol-ex ac ing
d ug me hyl-b-cyclodex in [85] o he pha macological ac i a ion
o he choles e ol deg ading enzyme CYP46 [84] in cellula and
animal models o Niemann-Pick ype C (OMIM 257220;
607625). Fluo escen -labelled chole a oxin subuni B un eiled
Fig. 3. Lipidomic analysis o he al e ed phospholipid composi ion in he ce ebel-
lum o a mouse model o Niemann Pick ype A (ASMko). Phospholipid composi ion
o he ce eb al co ex was analyzed by mass spec ome y in di e en mice (4 w
and 4 ASMko) a 4 mon hs o age. Di e ences in he le els o he indica ed
phospholipid species (PC; PE; PS; LPC; LPE; PC-O; PE-O) a e ep esen ed o each
indi idual mouse as a hea map exp essing he di e ences be ween he g oups a e
no maliza ion by using Me aboAnalys 5.0 so wa e.
Fig. 4. S udy o lysosomal al e a ions by mic oscopy. A: Immuno luo escence
agains he lysosomal memb ane p o ein LAMP1 in cul u ed p ima y neu ons om
w mice and om ASMko mice ha mimic Niemann Pick disease ype A. The
luo escence mic oscopy image illus a es he inc ease in lysosomal size due o lipid
accumula ion in he ASMko neu ons compa ed o w . Whi e lines delinea e he
neu ons. Scale ba = 10
l
m. B: Fluo escence mic oscopy image o he lyso acke
ed dye in cul u ed p ima y neu ons om w mice ea ed wi h ehicle o wi h
40
l
M sphingomyelin o inc ease he le els o his lipid as in he ASMko neu ons.
Lyso acke , which s ains acidic compa men s, shows a punc a e pa e n co e-
sponding o in ac lysosomes in he ehicle ea ed cells while ha ing a di use
cy osolic display in he SM ea ed cells indica ing lysosomal pe meabiliza ion.
Whi e lines delinea e he neu ons. Dapi in blue indica es nuclei. Scale ba = 10
l
m.
C: Elec on mic oscopy mic og aphs o mic oglia in WT and ASMko mice. The
images allow isualiza ion o expanded lysosomes and acuoles in he cy osol o
he ASMko mic oglia compa ed o he w . Scale ba = 2
l
m.
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
6
he e ec s o cyclodex in on GM1-ganglioside le els in cellula
models o gangliosidosis (OMIM 230500) [86]. BODIPY-labeled
C12 sphingomyelin was used as a subs a e o de ec acid sphin-
gomyelinase (ASM) ac i i y in ce eb ospinal luid and plasma a e
adeno-associa ed i al ec o -based gene he apy in he ASM
knock-ou mouse line ha models Niemann-Pick ype A (OMIM
257200) [87].
Fluo escence-based me hods ha e been key in he clinical diag-
nosis o LSDs, which hea ily elies on ei he de ec ion o he accu-
mula ion o molecules o iden i ica ion o de ec i e ac i i y o
enzymes in pa ien cells and luids. S aining o he au o luo escen
an ibio ic Filipin in pa ien ib oblas s has been adi ionally used
o he diagnosis o Niemann-Pick ype C (OMIM 257220;
607625) [88]. The luo escen -labelled ganglioside
sul o hodamine-GM1 has been used o de e mine he ac i i y o
hexosaminidase A in skin ib oblas s and whi e blood cells om
pa ien s o GM2-gangliosidosis (OMIM 272800), allowing
esea che s o dis inguish be ween he a ious sub ypes o his
LSD [89]. Ke a in sul a e-dependen luo escence has been used
o diagnose Mo quio A synd ome (OMIM 253000) om d ied blood
samples [90]. A simple me hod de e mined he enzyma ic ac i i y
o
a
-L- ucosidase by measu ing he luo escence quenching o
CdTe semiconduc o quan um do s in se um samples om ucosi-
dosis (OMIM 230000) pa ien s [91].
2.3. Elec on mic oscopy
Al hough less amenable han luo escence-based mic oscopy
me hods, elec on mic oscopy (EM) has also been widely used o
analyse he ul as uc u al changes ela ed o he deposi o non-
deg aded molecules wi hin he lysosomes and cy osol o cells om
pa ien s and LSD animal models (Fig. 4C). EM analysis o cells and
issues c ucially con ibu ed o he iden i ica ion o au ophagy
al e a ions as a common pa hological ea u e in many LSDs
[92,93]. Single-pa icle c yo-EM has yielded p o ein s uc u es
wi h inc easing le els o de ail in ecen yea s, and i can be
applied o explo e he dynamic con o ma ional changes o lysoso-
mal channels and p o eins. Al hough EM does no ha e he scaling
and ou pu o be adop ed o high h oughpu sc eening i has isen
as a complemen a y me hod o adi ional s uc u al echniques
and does show p omise in s uc u e-based d ug disco e y [94].
As EM has been used in he s udy o almos all LSDs, he e we
only wish o highligh a ew ele an examples, mainly in human
issue. In sphingolipidoses, a ecen comp ehensi e ul as uc u al
examina ion o he au opsy issue om a Niemann-Pick ype A
(OMIM 257200) pa ien epo ed signi ican accumula ion o sph-
ingomyelin e en in o gans ha did no show o e clinical mani-
es a ions [95]. Inclusions wi hin he cy oplasm o podocy es
we e obse ed in a pa ien ca ying uniden i ied gene a ia ion
associa ed wi h Fab y disease (OMIM 301500) [96]. EM ca ied
ou on skin biopsy samples om Fa be disease (OMIM 228000)
pa ien s ha bou ing a new mu a ion e ealed he p esence o
s acks o memb anes in enla ged lysosomes also known as zeb a
bodies [97].
In mucopolysaccha idoses, memb ane-bound acuoles and
deposi s o sul a ed acidic glycosaminoglycans we e de ec ed by
EM in biopsied muscles om a Hun e synd ome (OMIM 309900)
pa ien [98]. Ke a ocy es con aining acuoles and elec on-dense
lysosomes ull o p o eoglycans we e desc ibed in he co nea om
a pa ien o Ma o eaux-Lamy synd ome (OMIM 253200) [99].
Single-pa icle c yo-EM helped o esol e he s uc u al di e ences
be ween dis inc con o ma ions o he TRPML1 channel, and may
con ibu e o unde s and he po en ial mechanism o di e en
mu a ions in mucolipidosis ype IV (OMIM 252650) [100].
In glycop o einoses, EM echniques allowed he desc ip ion o
an accumula ion o e minal sialic acid a he non- educing end
o he suga chain in he b ain issue o a sialidosis pa ien (OMIM
256550) [101]. Dis ended lysosomes wi hin endo helial cells,
ib oblas s and his iocy es we e ound in he skin o a ucosidosis
pa ien (OMIM 230000) [102].
2.4. Op ogene ics
Op ogene ic me hods allow p ecise manipula ion o many cel-
lula ac i i ies wi h ligh [103]. Th ough he use o illumina ion
and he exp ession o pho op o eins, p o ein–p o ein in e ac ions
and biochemical pa hways can be con olled. Among he ad an-
ages o op ogene ic me hods a e he cell ype speci ici y, he high
speed and accu acy and less damage o issue. Among he main
challenges o op ogene ic implemen a ion in humans a e he
exp ession o adequa e amoun s o pho op o eins and he de elop-
men o low-hea ligh sou ces. Di e en op ogene ic s a egies
ha e been used in cellula and animal models o moni o and mod-
ula e ele an ea u es o lysosome physiology including hei pH
o mo ili y. As an example, ligh ac i a ion o lyso-pHoenix, a
usion p o ein made up o he ligh -ac i a ed p o on pump A ch3,
he pH-sensi i e GFP a ian pHluo in and he lysosomal- a ge ing
CD63 p o ein, caused a apid d op in lysosomal pH [104]. This
p obe enables he ex e nal con ol o lysosomal pH and can be
applied o di ec ly assess he acid-dependen ac i i y o lysosomal
p o eases and he ole o acidi ica ion in LSD pa hology. Op oge-
ne ic s a egies ha e also been used o con ol he anspo and
dis ibu ion o lysosomes. Upon ligh -induced ec ui men o
molecula mo o s such as kinesin o dynein, eposi ioning o hese
o ganelles in cells has been achie ed [105]. Op ogene ic modules
Fig. 5. P ima y cell cul u es o s udy lipid al e a ions in LSDs. A: Su ace s aining in
non-pe meabilized p ima y cul u ed neu ons om WT mice and ASMko mice,
which mimic Niemann Pick disease Type A, using an an ibody agains he oxin-
de i ed pep ide Lysenin ha speci ically binds sphingomyelin. This kind o
expe imen showed ha in ASMko neu ons sphingomyelin accumula ion no only
occu s in he lysosomes bu also a he plasma memb ane. Scale ba = 5
l
m. B:
Lysenin s aining in pe meabilized cul u ed ib oblas ob ained om a con ol
subjec and a Niemann Pick disease ype A pa ien e idencing he in acellula
accumula ion o sphingomyelin in he pa ien cell. Whi e ci cles indica e he nuclei.
Scale ba = 10
l
m.
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
7
based on ligh -sensi i e dime ize s ha e been used o ansloca e
mTORC1 o lysosomes upon pho oac i a ion [106]. Pho o oxic p o-
eins ha e been a ge ed o he cy oplasmic su ace o lysosomes
by usion wi h he small GTPase Rab7. This s a egy has been used
o induce lysosomal memb ane pe meabiliza ion [107] o oxida-
i e s ess [108] upon illumina ion.
2.5. Liposome-based echniques
Liposome-based echniques ha e con ibu ed o bo h cha ac e -
iza ion and ea men o pa hology in LSD models. Liposomes a e
sphe ical pa icles made up o di e en ypes o lipids ha o gan-
ise o o m one o mo e lipid bilaye s. They can be syn hesized by
di e en means [109] including he mechanical me hods o hin
ilm hyd a ion [110] o sol en dispe sion [111]. The b oad appli-
ca ion o liposomes mainly de i es om hei capaci y o encapsu-
la e di e en ypes o compounds o gene ic ec o s [112].
Liposomes can deli e hei ca go o almos all o gans and issues.
This includes he b ain when s a egies like coa ing o conjuga ion
wi h an ibodies a e used o help liposomes c oss he blood–b ain
ba ie (BBB) [113]. Se e al key ea u es mus be de e mined
in i o be o e applying liposomes in clinical esea ch se ings,
including solubili y, s abili y, ca go up ake, abili y o c oss
endo helial ba ie s and BBB s uc u es, and e ec i e deli e y o
ca go in o he desi ed cell/o ganelle. We e e o he a icles in his
issue by D s. Ven osa, Ceccini and Mu o o excellen e iews on
he use o liposomes and o he nanopa icles in LSD he apy.
In addi ion o he apeu ics, liposome-based echniques ha e
been used as ools o s udy LSD pa hological mechanisms and o
c ea e LSD models. In sphingolipidoses, a de e gen - ee liposomal
assay unco e ed he egula o y oles o dis inc lipid classes on he
hyd olysis o memb ane-bound glucosylce amide by he lysosomal
b-glucoce eb osidase in Gauche disease (OMIM 230800) models
[114]. Recons i u ion o GM2 ca abolism a liposomal su aces
was used as a model o s udy he e ec o he accumula ion o di -
e en memb ane lipids on ganglioside me abolism and gangliosi-
dosis [115].
In glycop o einoses, liposomes loaded wi h glycolipids we e
used o demons a e he de icien deg ada ion o hese lipids by
skin ib oblas s om pa ien s o Schindle disease (OMIM
609241) [116].
3. Cellula models in LSD esea ch
Cellula models ha e been commonly used in LSD esea ch.
F om adi ional monocul u ed cells in isola ion o cu ing-edge
echnologies leading o he gene a ion o mul i-lineage cell pla -
o ms, hese echniques ha e acili a ed ou unde s anding o
pa hological molecula mechanisms and d i en he disco e y o
d ug- and cell-based he apies.
3.1. P ima y cell cul u es
P ima y cell cul u e is de ined as he ex i o cul u e o cells
eshly ob ained om a issue. The main ad an ages o p ima y
cells a e hei ai h ul ansc ip omic and p o eomic p o ile and
close p oximi y o physiological unc ion and esponse. Some
d awbacks a e hei equi emen s o speci ic subs a es and nu i-
en s and he acquisi ion o a senescen pheno ype, which leads o
an i e e sible cell cycle a es ha limi s he amoun o ma e ial
ha can be ob ained. This p e en s he use o p ima y cul u es
o la ge-scale disease modelling and d ug disco e y p ojec s.
P ima y cul u es o ib oblas s om pa ien -de i ed skin biop-
sies ha e been b oadly used in LSDs, no only o unde s and dis-
ease mechanisms bu also o diagnos ic pu poses (Fig. 4B). In
mucopolysaccha idoses, ansc ip omic s udies on ib oblas cul-
u es de i ed om pa ien s su e ing om all ypes o hese dis-
eases iden i ied dys egula ion o apop osis- ela ed genes [117].In
glycop o einoses, he use o p ima y cul u ed ib oblas s om
pa ien s su e ing om sialidosis (OMIM 256550) and galac osiali-
dosis (OMIM 256540) demons a ed impai ed elas ogenesis ha
could be e e ed by ansduc ion wi h neu aminidase-1 cDNA o
a e ea men wi h bac e ial sialidase [118]. Filipin s aining o
de ec choles e ol accumula ion in p ima y ib oblas s om
Niemann-Pick ype C (OMIM 257220; 607625) pa ien s has been
a key diagnos ic ool o decades [119].
The use o p ima y neu onal cul u es has been pa icula ly
in o ma i e o he neu ological pheno ypes in LSDs. Bo h mono-
cul u es and co-cul u es o p ima y b ain cells ob ained om
mouse models ha e been used. In sphingolipidoses, p ima y cul-
u ed neu ons om ASM knock-ou mice, which mimic
Niemann-Pick ype A (OMIM 257200) (Fig. 4A), un eiled al e -
a ions in au ophagy [72], calcium homeos asis [80] and synap ic
ea u es [120,121]. In mucopolysaccha idoses, co-cul u es o p i-
ma y b ain cells om mice lacking sul amidase (which mimic San-
ilippo synd ome (OMIM 252900)) e ealed he highes
accumula ion o hepa an sul a e in as ocy es [122].
3.2. Immo alized cell lines
Immo alized cell lines a e gene a ed by na u ally occu ing o
induced mu a ions ha a oid senescence and allow cells o keep
di iding. They can be used in esea ch ins ead o p ima y cul u es,
p o iding pu e popula ions o cells and mo e ep oducible esul s.
Among o he ad an ages, hey a e cos e ec i e, deli e an almos
unlimi ed supply o ma e ial and bypass he e hical conce ns asso-
cia ed wi h he use o animal and human issue. Howe e , due o
he gene ic manipula ion ha is equi ed o gene a e hem,
immo alized cell lines may show al e ed pheno ype when com-
pa ed o p ima y cul u es. Se ial passage can also cause geno ypic
and pheno ypic a ia ions and con amina ion wi h mycoplasma is
common [123]. Despi e hese d awbacks, immo alized cell lines
ha e been ins umen al in LSD esea ch. They may exp ess speci ic
lysosomal enzymes o be used as he apeu ic ools. Mo eo e , he
clus e ed egula ly in e spaced sho palind omic epea s
(CRISPR)/CRISPR-associa ed p o ein (Cas) sys em, i s ly disco e ed
as a de ence sys em in bac e ia and a chaea [124], acili a es gen-
ome edi ing and is a powe ul ool [125] ha is inc easingly being
used o gene a e LSD disease modelling cell lines. O pa icula el-
e ance in he LSD con ex a e immo alized cells gene a ed a e
ans o ma ion o pa ien -de i ed p ima y cells by, o example,
hTERT o SV40 la ge T an igen echnology [126]. Compa isons
among single-pa ien de i ed cell lines a e use ul o unde s and
he a iable pheno ype o LSD pa ien s, which o en depends on
he speci ic mu a ions hey ca y.
In sphingolipidoses, a Chinese hams e o a y cell line o e ex-
p essing human GM2A was gene a ed o pu i y he sec e ed
enzyme and s udy i s up ake by ib oblas s de i ed om Sandho
disease (OMIM 268800) pa ien s [127]. A human monocy ic THP-1
cell line, in which he GBA1 gene was edi ed, was used o model
Gauche disease (OMIM 230800) [128]. GLA gene knock-ou
HEK-293T cells we e gene a ed by he CRISPR/Cas9 me hodology
o s udy he cellula pha macokine ics o ecombinan human
a
-
galac osidase A [129]. The he apeu ic po en ial o gene edi ed
neu al s em cells o e exp essing galac osylce amidase was shown
in K abbe disease (OMIM 245200) [130]. An NPC1 knock-ou HeLa
cell line was gene a ed by CRISPR/Cas9 me hodology o s udy
Niemann-Pick ype C (OMIM 257220) [131].
In mucopolysaccha idoses, ca hepsin B leakage om lysosomes
o he cy oplasm was epo ed in a Hun e synd ome (OMIM
309900)-de i ed neu onal cell line [132]. NAGLU-de icien and
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
8
HGSNAT-mu a ed cell lines modelling San ilippo synd ome ypes B
(OMIM 252920) and C (OMIM 252930), espec i ely, we e gene -
a ed by CRISPR/Cas9 echnology [133,134].
In glycop o einoses, RAG and LM/TK cell lines con ibu ed o he
ch omosomal assignmen o wo genes associa ed wi h
neu aminidase-de iciency diso de s [135].
3.3. Induced plu ipo en s em cells (iPSCs)
iPSCs a e gene a ed by ep og amming o soma ic cells h ough
he manipula ion o speci ic ansc ip ion ac o s and exhibi he
mo phology, g ow h p ope ies and ma ke genes o emb yonic
s em cells [136,137]. iPSCs can be ob ained om di e en esea ch
animals, and e en om human biopsies. Human iPSCs (hiPSCs)
ha bou na u al-disease causing mu a ions and allow s udy o
he ole o gene ic backg ound in disease p og ession and se e i y.
Impo an ly, hey can be di e en ia ed o p oduce cellula lineages
ha canno be ob ained di ec ly om pa ien s, such as neu ons o
neu al s em cells. The e o e, hey a e ins umen al o s udy he
neu ological pheno ypes ha cha ac e ize many LSDs (Table 2).
Gi en hei inhe en pa ien speci ici y, hiPSC-de i ed cell ypes
may be key o de elop pe sonalized d ug ea men s [163]. E o s
a e cu en ly unde way o educe he a iabili y o di ec ed hiPSC
di e en ia ion, which can lead o popula ions o mul iple cell ypes
ha a ise spon aneously [164].
Use o human iPSCs in LSD esea ch has been ex ensi ely
e iewed in [165]. He e, we p o ide only some examples ha high-
ligh hei u ili y. In sphingolipidoses, oxida i e s ess and p esy-
nap ic dys unc ion we e desc ibed in neu ons c ea ed om iPSCs
ob ained om skin-de i ed ib oblas o pa ien s wi h Tay-Sachs
disease (OMIM 272800) [142]. Pa ien -de i ed iPSCs and gene-
edi ing echnology we e used o s udy he ca diac- ela ed molecu-
Table 2
Lis o iPSCs and o ganoids gene a ed o LSD esea ch. hiPSCs: Human-induced plu ipo en s em cells. iPSCs: Induced plu ipo en s em cells. ND: No desc ibed.
Disease iPSCs O ganoids
Sphingolipidoses
GM1 gangliosidosis Neu ons di e en ia ed om
pa ien -de i ed hiPSCs [138]
Ce eb al o ganoids om iPSCs [139]
Sandho disease iPSCs de i ed om mouse model
neu al s em cells [140]
Ce eb al o ganoids om pa ien -de i ed hiPSCs [141]
Tay-Sachs disease hiPSCs om pa ien -de i ed
ib oblas s [142]
Ce eb al o ganoids om pa ien -de i ed hiPSCs [141]
Fab y disease hiPSCs om pa ien -de i ed
ib oblas s [143]
Me ach oma ic leukodys ophy hiPSCs line om pa ien -de i ed
ib oblas s [144]
A i icially iPSCs-de i ed whole-b ain o ganoid [145]
K abbe disease hiPSCs line om pa ien -de i ed
ib oblas s [146]
ND
Niemann-Pick A/B disease hiPSCs line om ype A pa ien -
de i ed de mal ib oblas s [147]
hiPSCs line om ype B pa ien -
de i ed de mal ib oblas s [148]
ND
Fa be disease hiPSCs line om pa ien -de i ed
ib oblas s [149]
ND
Gauche disease hiPSCs line om pa ien -de i ed
ib oblas s [150]
ND
Mucopolysaccha idoses
Hun e synd ome hiPSCs om pa ien -de i ed
de mal ib oblas s [151]
ND
Hu le -Scheie synd ome hiPSCs om pa ien -de i ed
ib oblas s [152]
ND
San ilippo synd ome C HGSNAT-mu a ed cell lines om
heal hy hIPSCs [134]
ND
San ilippo synd ome B hiPSCs om pa ien -de i ed
pe iphe al blood mononuclea
cells [153]
ND
Sly disease hiPSCs om pa ien -de i ed
ib oblas s [154]
ND
Mo quio A synd ome hiPSCs om pa ien -de i ed
de mal ib oblas s [155]
ND
Glycop o einoses/Oligosaccha idoses
Sialidosis ype I IPSCs-de i ed neu al cells
gene a ed om pa ien -de i ed
ib oblas s [156]
ND
Sialidosis ype II IPSCs-de i ed neu al cells
gene a ed om pa ien -de i ed
ib oblas s [156]
ND
Non-enzyma ic LSDs
Niemann Pick ype C Neu ons di e en ia ed om
i
3
Neu on iPSCs [157]
hiPSCs om pa ien -de i ed
ib oblas s [158]
Ce eb al o ganoids om pa ien -de i ed hiPSCs [159]
Danon disease hiPSCs om pa ien -de i ed
pe iphe al blood mononuclea
cells [160]
ND
Ba en disease Gene co ec ion s udies in
hiPSCS om pa ien -de i ed
de mal ib oblas s [161]
Ce eb al o ganoids om pa ien -de i ed hiPSCs [162]
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
9
Tiemeije , S.W. Ha dwick, D.Y. Chi gadze, G. Mu shudo , A.R. A icescu, S.H.W.
Sche es, Single-pa icle c yo-EM a a omic esolu ion, Na u e 587 (2020)
152–156.
[95] B.L. Thu be g, Au opsy pa hology o in an ile neu o isce al ASMD (Niemann-
Pick Disease ype A): Clinicopa hologic co ela ions o a case epo , Mol.
Gene . Me ab. Rep. 24 (2020) 100626.
[96] A.R. Vanga, S.A. Sch ie Ve gano, J. Kowalewska, T.R. McCune, P e iously
uniden i ied gene a ia ion associa ed wi h ab y disease: he impac on one
amily, Case Repo s in Neph ology (2020, 2020) 8899703.
[97] A.K. Ched awi, Z.N. Al-Hassnan, M. Al-Muhaizea, D. Colak, B. Al-Younes, A.
Albakhee , S. Tulba, N. Kaya, No el V97G ASAH1 mu a ion ound in Fa be
disease pa ien s: unique appea ance o he disease wi h an in e media e
se e i y, and ma ked ea ly in ol emen o cen al and pe iphe al ne ous
sys em, B ain & De . 34 (2012) 400–404.
[98] S. Wakai, R. Minami, K. Kameda, M. Okabe, M. Nagaoka, S. Annaka, Skele al
muscle in ol emen in mucopolysaccha idosis ype IIA: se e e ype o
Hun e synd ome, Pedia . Neu ol. 4 (1988) 178–180.
[99] S. Akh a , A. Tullo, B. Ca e son, J.R. Da ies, K. Benne , K.M. Meek, Clinical and
mo phological ea u es including exp ession o be aig-h3 and ke a an
sulpha e p o eoglycans in Ma o eaux-Lamy synd ome ype B and in no mal
co nea, B . J. Oph halmology 86 (2002) 147–151.
[100] S. Zhang, N. Li, W. Zeng, N. Gao, M. Yang, C yo-EM s uc u es o he
mammalian endo-lysosomal TRPML1 channel elucida e he combined
egula ion mechanism, P o ein & Cell 8 (2017) 834–847.
[101] T. Uchiha a, K. Ohashi, M. Ki agawa, M. Ku a a, A. Nakamu a, K. Hi okawa, T.
Kasuga, T. Kobayashi, Sialidosis ype I ca ying V217M/G243R mu a ions in
lysosomal sialidase: an au opsy s udy demons a ing e minal sialic acid in
lysosomal lamella inclusions and ce ebella dysplasia, Ac a Neu opa hol.
119 (2010) 135–145.
[102] C. Fleming, A. Rennie, M. Fallow ield, P.M. McHen y, Cu aneous
mani es a ions o ucosidosis, B . J. De ma ology 136 (1997) 594–597.
[103] K. Deisse o h, Op ogene ics, Na . Me hods 8 (2011) 26–29.
[104] B.R. Ros , F. Schneide , M.K. G auel, C. Wozny, C. Ben z, A. Blessing, T.
Rosenmund, T.J. Jen sch, D. Schmi z, P. Hegemann, C. Rosenmund,
Op ogene ic acidi ica ion o synap ic esicles and lysosomes, Na . Neu osci.
18 (2015) 1845–1852.
[105] L. Duan, D. Che, K. Zhang, Q. Ong, S. Guo, B. Cui, Op ogene ic con ol o
molecula mo o s and o ganelle dis ibu ions in cells, Chem. Biol. 22 (2015)
671–682.
[106] D. Khamsing, S. Leb un, I. Fange , N. La oche e, C. Tou ain, V. de Sa s, M.
B uns ein, M. Oheim, D. Ca el, F. Da chen, C. Desnos, A ole o BDNF- and
NMDAR-induced lysosomal ec ui men o mTORC1 in he egula ion o
neu onal mTORC1 ac i i y, Mol. B ain 14 (2021) 112.
[107] A.P. Ryumina, E.O. Se eb o skaya, D.B. S a o e o , O.A. Zlobo skaya, A.S.
Shcheglo , S.A. Lukyano , K.A. Lukyano , Lysosome-associa ed miniSOG as a
pho osensi ize o mammalian cells, Bio echniques 61 (2016) 92–94.
[108] E.O. Se eb o skaya, A.P. Ryumina, M.E. Boulina, M.V. Shi mano a, E.V.
Zagayno a, E.A. Bogdano a, S.A. Lukyano , K.A. Lukyano , Pho o oxic
e ec s o lysosome-associa ed gene ically encoded pho osensi ize
Kille Red, J. Biomed. Op . 19 (2014) 071403.
[109] C.G. Sion o ou, G.P. Nikoleli, D.P. Nikolelis, S.K. Ka ape is, A i icial lipid
memb anes: pas , p esen , and u u e, Memb anes 7 (2017).
[110] A.D. Bangham, M.M. S andish, J.C. Wa kins, Di usion o uni alen ions ac oss
he lamellae o swollen phospholipids, J. Mol. Biol. 13 (1965) 238–252.
[111] Y. Huang, Z. Liu, R. Bo, J. Xing, L. Luo, S. Zhen, Y. Niu, Y. Hu, J. Liu, Y. Wu, D.
Wang, The enhanced immune esponse o PCV-2 accine using Rehmannia
glu inosa polysaccha ide liposome as an adju an , In . J. Biol. Mac omol. 86
(2016) 929–936.
[112] D.B. Viei a, L.F. Gama a, Ge ing in o he b ain: liposome-based s a egies o
e ec i e d ug deli e y ac oss he blood-b ain ba ie , In . J. Nanomed. 11
(2016) 5381–5414.
[113] M.L. Pinzon-Daza, I. Campia, J. Kopecka, R. Ga zon, D. Ghigo, C. Rigan i,
Nanopa icle- and liposome-ca ied d ugs: new s a egies o ac i e a ge ing
and d ug deli e y ac oss blood-b ain ba ie , Cu . D ug Me ab. 14 (2013)
625–640.
[114] M. Abdul-Hammed, B. B eiden, G. Schwa zmann, K. Sandho , Lipids egula e
he hyd olysis o memb ane bound glucosylce amide by lysosomal be a-
glucoce eb osidase, J. Lipid Res. 58 (2017) 563–577.
[115] S. Anheuse , B. B eiden, K. Sandho , Memb ane lipids and hei deg ada ion
compounds con ol GM2 ca abolism a in alysosomal luminal esicles, J.
Lipid Res. 60 (2019) 1099–1111.
[116] B. As aw, D. Schindle , J. Led ino a, B. Ce ny, F. Smid, E. Conzelmann,
Deg ada ion o blood g oup A glycolipid A-6-2 by no mal and mu an human
skin ib oblas s, J. Lipid Res. 39 (1998) 1768–1780.
[117] J. B okowska, K. Pie zynowska, L. Ga ke, E. Rin z, G. Weg zyn, Exp ession o
genes in ol ed in apop osis is dys egula ed in mucopolysaccha idoses as
e ealed by pilo ansc ip omic analyses, Cell Biol. In . 45 (2021) 549–557.
[118] A. Hinek, A.V. Pshezhe sky, M. on I zs ein, B. S a che , Lysosomal sialidase
(neu aminidase-1) is a ge ed o he cell su ace in a mul ip o ein complex
ha acili a es elas ic ibe assembly, J. Biol. Chem. 281 (2006) 3698–3710.
[119] D. Si a ska, A. Lugowska, Labo a o y diagnosis o he Niemann-Pick ype C
disease: an inhe i ed neu odegene a i e diso de o choles e ol me abolism,
Me ab. B ain Dis. 34 (2019) 1253–1260.
[120] P.G. Camole o, H. Va a, L. Mo ando, E. Connell, F.P. Ma le o, M. Gius e o, M.
Sassoe-Pogne o, P.P. Van Veldho en, M.D. Ledesma, Synap ic esicle
docking: sphingosine egula es syn axin1 in e ac ion wi h Munc18, PLoS
ONE 4 (2009) e5310.
[121] A.I. A oyo, P.G. Camole o, L. Mo ando, M. Sassoe-Pogne o, M. Gius e o, P.P.
Van Veldho en, E.H. Schuchman, M.D. Ledesma, Pha macological e e sion o
sphingomyelin-induced dend i ic spine anomalies in a Niemann Pick disease
ype A mouse model, EMBO Mol. Med. 6 (2014) 398–413.
[122] C.A. Dwye , S.L. Scudde , Y. Lin, L.E. Dozie , D. Phan, N.J. Allen, G.N. Pa ick, J.
D. Esko, Neu ode elopmen al changes in exci a o y synap ic s uc u e and
unc ion in he ce eb al co ex o san ilippo synd ome IIIA mice, Sci. Rep. 7
(2017) 46576.
[123] G. Kau , J.M. Du ou , Cell lines: Valuable ools o useless a i ac s,
Spe ma ogenesis 2 (2012) 1–5.
[124] F.J. Mojica, G. Juez, F. Rod iguez-Vale a, T ansc ip ion a di e en salini ies o
Halo e ax medi e anei sequences adjacen o pa ially modi ied Ps I si es,
Mol. Mic obiol. 9 (1993) 613–621.
[125] R. Ricci, G. Colasan e, CRISPR/dCas9 as a he apeu ic app oach o
neu ode elopmen al diso de s: inno a ions and limi a ions compa ed o
adi ional s a egies, De . Neu osci. (2021) 1–9.
[126] X. Wu, S. Wang, M. Li, J. Li, J. Shen, Y. Zhao, J. Pang, Q. Wen, M. Chen, B. Wei, P.
J. Kaboli, F. Du, Q. Zhao, C.H. Cho, Y. Wang, Z. Xiao, X. Wu, Condi ional
ep og amming: nex gene a ion cell cul u e, Ac a Pha maceu ica Sinica. B 10
(2020) 1360–1381.
[127] K. Ki akaze, C. Tasaki, Y. Tajima, T. Hi okawa, D. Tsuji, H. Saku aba, K. I oh,
Combined eplacemen e ec s o human modi ied be a-hexosaminidase B
and GM2 ac i a o p o ein on GM2 gangliosidoses ib oblas s, Biochem.
Biophys. Rep. 7 (2016) 157–163.
[128] E. Pa an, M. O mazabal, P. Pe uzzo, E. Vaena, P. Rozen eld, A. Da dis, CRISPR/
Cas9 edi ing o gauche disease modelling, In . J. Mol. Sci. 21 (2020).
[129] H.Y. Song, H.C. Chiang, W.L. Tseng, P. Wu, C.S. Chien, H.B. Leu, Y.P. Yang, M.L.
Wang, Y.J. Jong, C.H. Chen, W.C. Yu, S.H. Chiou, Using CRISPR/Cas9-media ed
GLA gene knockou as an in i o d ug sc eening model o ab y disease, In .
J. Mol. Sci. 17 (2016).
[130] D.P. De e , S.G. Scha enbe g, J. Cama ena, E.J. Kildebeck, J.T. Cla k, R.M.
Ma in, R.O. Bak, Y. Tang, M. Dohse, J.A. Bi gmeie , K.A. Jagadeesh, G.
Beje ano, A. Tsukamo o, N. Gomez-Ospina, N. Uchida, M.H. Po eus, CRISPR/
Cas9 genome enginee ing in eng a able human b ain-de i ed neu al s em
cells, iScience 15 (2019) 524–535.
[131] X. Du, I. Lukman a a, H. Yang, CRISPR/Cas9-media ed gene a ion o niemann-
Pick C1 knockou cell line, Me hods Mol. Biol. 1583 (2017) 73–83.
[132] A.S. Azambuja, L.N. Pimen el-Ve a, E.A. Gonzalez, E. Pole o, C.V. Pinhei o, U.
Ma e, R. Giugliani, G. Baldo, E idence o in lammasome ac i a ion in he
b ain o mucopolysaccha idosis ype II mice, Me ab. B ain Dis. 35 (2020)
1231–1236.
[133] N. Bene o, M. Coza , L. Go , L. Pacheco, L. Vilageliu, D. G inbe g, I. Canals,
Gene a ion o wo NAGLU-mu a ed homozygous cell lines om heal hy
induced plu ipo en s em cells using CRISPR/Cas9 o model San ilippo B
synd ome, S em Cell Res. 42 (2020) 101668.
[134] N. Bene o, M. Coza , M. Ga cia-Mo an , E. C eus-Bachille , L. Vilageliu, D.
G inbe g, I. Canals, Gene a ion o wo compound he e ozygous HGSNAT-
mu a ed lines om heal hy induced plu ipo en s em cells using CRISPR/Cas9
o model San ilippo C synd ome, S em Cell Res. 41 (2019) 101616.
[135] O.T. Muelle , W.M. Hen y, L.L. Haley, M.G. Bye s, R.L. Eddy, T.B. Shows,
Sialidosis and galac osialidosis: ch omosomal assignmen o wo genes
associa ed wi h neu aminidase-de iciency diso de s, PNAS 83 (1986) 1817–
1821.
[136] K. Takahashi, S. Yamanaka, Induc ion o plu ipo en s em cells om mouse
emb yonic and adul ib oblas cul u es by de ined ac o s, Cell 126 (2006)
663–676.
[137] K. Takahashi, K. Tanabe, M. Ohnuki, M. Na i a, T. Ichisaka, K. Tomoda, S.
Yamanaka, Induc ion o plu ipo en s em cells om adul human ib oblas s
by de ined ac o s, Cell 131 (2007) 861–872.
[138] R. Kajiha a, T. Numakawa, H. Odaka, Y. Yaginuma, N. Fusaki, T. Okumiya, H.
Fu uya, S. Inui, T. E a, No el d ug candida es imp o e ganglioside
accumula ion and neu al dys unc ion in GM1 gangliosidosis models wi h
au ophagy ac i a ion, S em Cell Rep. 14 (2020) 909–923.
[139] Y.L. La ou , R. Yoon, S.E. Thomas, C. G an , C. Li, M. Sena-Es e es, M.L. Allende,
R.L. P oia, C.J. Ti , Human GLB1 knockou ce eb al o ganoids: A model
sys em o es ing AAV9-media ed GLB1 gene he apy o educing GM1
ganglioside s o age in GM1 gangliosidosis, Mol. Gene . Me ab. Rep. 21 (2019)
100513.
[140] Y. Ogawa, K. Kaizu, Y. Yanagi, S. Takada, H. Saku aba, K. Oishi, Abno mal
di e en ia ion o Sandho disease model mouse-de i ed mul ipo en s em
cells owa d a neu al lineage, PLoS ONE 12 (2017) e0178978.
[141] M.L. Allende, E.K. Cook, B.C. La man, A. Nugen , J.M. B ady, D. Golebiowski, M.
Sena-Es e es, C.J. Ti , R.L. P oia, Ce eb al o ganoids de i ed om Sandho
disease-induced plu ipo en s em cells exhibi impai ed neu odi e en ia ion,
J. Lipid Res. 59 (2018) 550–563.
[142] K. Ma sushi a, T. Numakawa, H. Odaka, R. Kajiha a, M. Soga, S. Ozasa, K.
Nakamu a, H. Mizu a, T. E a, P esynap ic dys unc ion in neu ons de i ed
om ay-sachs iPSCs, Neu oscience 414 (2019) 128–140.
[143] M.J. Bi ke , S. Raibaud, M. Le ie i, A.D. Adamson, V. Le ang, P. Ce ello, N.
Redon, G. Re , S. Viale, B. Wang, B. Bi on, J.C. Guillemo , V. Mikol, J.P. Leona d,
N.A. Hanley, C. O sini, J.M. I ie , A human s em cell model o ab y disease
implica es LIMP-2 accumula ion in ca diomyocy e pa hology, S em Cell Rep.
13 (2019) 380–393.
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
16

[144] K.P. Kim, J. Yoon, J. Kim, A. Ropke, B. Shin, D. Wook Han, B. G ebe , H.R.
Schole , Gene a ion o a human iPSC line (MPIi007-A) om a pa ien wi h
Me ach oma ic leukodys ophy, S em Cell Res. 48 (2020) 101993.
[145] S. Esmail, W.R. Dan e , A i icially induced plu ipo en s em cell-de i ed
whole-b ain o ganoid o modelling he pa hophysiology o me ach oma ic
leukodys ophy and d ug epu posing, Biomedicines 9 (2021).
[146] W. Wang, Y.F. L , Y.J. Zhang, W.J. Dong, Y. Zhang, Gene a ion o a human
induced plu ipo en s em cell line PUMCi001-A om a pa ien wi h K abbe
disease, S em Cell Res. 48 (2020) 101937.
[147] A. Bask ield, R. Li, J. Bee s, J. Zou, C. Liu, W. Zheng, An induced plu ipo en
s em cell line (TRNDi009-C) om a Niemann-Pick disease ype A pa ien
ca ying a he e ozygous p. L302P (c.905 T>C) mu a ion in he SMPD1 gene,
S em Cell Res. 38 (2019) 101461.
[148] A. Bask ield, R. Li, J. Bee s, J. Zou, C. Liu, W. Zheng, Gene a ion o an induced
plu ipo en s em cell line (TRNDi004-I) om a Niemann-Pick disease ype B
pa ien ca ying a he e ozygous mu a ion o p. L43_A44delLA in he SMPD1
gene, S em Cell Res. 37 (2019) 101436.
[149] B.M. B ooks, C.D. Yeh, J. Bee s, C. Liu, Y.S. Cheng, K. Go shko , J. Zou, W.
Zheng, C.Z. Chen, Gene a ion o an induced plu ipo en s em cell line
(TRNDi030-A) om a pa ien wi h Fa be disease ca ying a homozygous p.
Y36C (c. 107 A>G) mu a ion in ASAH1, S em Cell Res., 53 (2021) 102387.
[150] Q. Zhao, Y. Cui, J. Wang, L. Shi, Z. Qi, J. Luan, Y. Zhang, X. Zhou, J. Han,
De elopmen o a human iPSC line (SMBCi004-A) om a pa ien wi h
Gauche disease, S em Cell Res. 48 (2020) 101989.
[151] J. Hong, M. Xu, R. Li, Y.S. Cheng, J. Kouzne so a, J. Bee s, C. Liu, J. Zou, W.
Zheng, Gene a ion o an induced plu ipo en s em cell line (TRNDi008-A)
om a Hun e synd ome pa ien ca ying a hemizygous 208insC mu a ion in
he IDS gene, S em Cell Res. 37 (2019) 101451.
[152] S. Li o, P. Bu da, M. Baumga ne , F. Sloan-Bena, Z. Tancos, J. Kobolak, A.
Dinnyes, K.H. K ause, A. Ma eyn, Gene a ion o human induced plu ipo en
s em cell line UNIGEi001-A om a 2-yea s old pa ien wi h
Mucopolysaccha idosis ype IH disease, S em Cell Res. 41 (2019) 101604.
[153] J. Guan, G. Tian, R. Dong, H. Zhang, X. Yang, Y. Li, Z. Gai, Y. Liu, Induced
plu ipo en s em cell line (SDQLCHi041-A) om a male pa ien wi h
mucopolysaccha idosis ype IIIB, S em Cell Res. 52 (2021) 102212.
[154] T.A. G i in, H.C. Ande son, J.H. Wol e, Ex i o gene he apy using pa ien
iPSC-de i ed NSCs e e ses pa hology in he b ain o a homologous mouse
model, S em Cell Rep. 4 (2015) 835–846.
[155] R. Li, A. Bask ield, J. Bee s, J. Zou, C. Liu, C.J. Almeciga-Diaz, W. Zheng,
Gene a ion o an induced plu ipo en s em cell line (TRNDi005-A) om a
Mucopolysaccha idosis Type IVA (MPS IVA) pa ien ca ying compound
he e ozygous p.R61W and p.WT405del mu a ions in he GALNS gene, S em
Cell Res., 36 (2019) 101408.
[156] H. Odaka, T. Numakawa, M. Soga, J. Kido, S. Ma sumo o, R. Kajiha a, T.
Okumiya, N. Tani, Y. Tanoue, T. Fukuda, H. Fu uya, T. Inoue, T. E a, An iPSC-
based neu al model o sialidosis unco e s glycoly ic impai men -causing
p esynap ic dys unc ion and de egula ion o Ca(2+) dynamics, Neu obiol. Dis.
152 (2021) 105279.
[157] A.V. P abhu, I. Kang, R. De Pace, C.A. Wassi , H. Fujiwa a, P. Kell, X. Jiang, D.S.
O y, J.S. Boni acino, M.E. Wa d, F.D. Po e , A human iPSC-de i ed inducible
neu onal model o Niemann-Pick disease, ype C1, BMC Biol. 19 (2021) 218.
[158] C. Volkne , M. Lied ke, J. Pe e s, J. Lukas, H.M. Escoba , G. Knuebel, J.
Bulle diek, C. Holzmann, A. He mann, M.J. F ech, Gene a ion o an iPSC line
(AKOSi004-A) om ib oblas s o a emale adul NPC1 pa ien , ca ying he
compound he e ozygous mu a ion p.Val1023Se s*15/p.Gly992A g and o an
iPSC line (AKOSi005-A) om a emale adul con ol indi idual, S em Cell Res.
50 (2020) 102127.
[159] S.E. Lee, N. Shin, M.G. Kook, D. Kong, N.G. Kim, S.W. Choi, K.S. Kang, Human
iNSC-de i ed b ain o ganoid model o lysosomal s o age diso de in
Niemann-Pick disease ype C, Cell Dea h Dis. 11 (2020) 1059.
[160] X. Zhu, J. Wang, H. Chong, J. Wang, D. Wang, Gene a ion o induced
plu ipo en s em cells (NJDTHi001-A) om a Danon disease child wi h
mu a ion o c.467 T > G in LAMP2 gene, S em Cell Res. 49 (2020) 101994.
[161] X. Zhang, D. Zhang, J.A. Thompson, S.C. Chen, Z. Huang, L. Jennings, T.L.
McLa en, T.M. Lamey, J.N. De Roach, F.K. Chen, S. McLenachan, Gene
co ec ion o he CLN3 c.175G>A a ian in pa ien -de i ed induced
plu ipo en s em cells p e en s pa hological changes in e inal o ganoids,
Mol. Gene . Genomic Med. 9 (2021) e1601.
[162] G. Gomez-Gi o, J. A ias-Fuenzalida, J. Ja azo, D. Zeuschne , M. Ali, N.
Possemis, S. Bolognin, R. Halde , C. Jage , W.F.E. Kupe , P.M. an Hassel , H.
Zaeh es, A. Del Sol, H. an de Pu en, H.R. Schole , J.C. Schwambo n, Synapse
al e a ions p ecede neu onal damage and s o age pa hology in a human
ce eb al o ganoid model o CLN3-ju enile neu onal ce oid lipo uscinosis,
Ac a Neu opa hologica Commun. 7 (2019) 222.
[163] A. Sha ma, S. Sances, M.J. Wo kman, C.N. S endsen, Mul i-lineage human
iPSC-de i ed pla o ms o disease modeling and d ug disco e y, Cell S em
Cell 26 (2020) 309–329.
[164] S. Sances, L.I. B uijn, S. Chand an, K. Eggan, R. Ho, J.R. Klim, M.R. Li esey, E.
Low y, J.D. Macklis, D. Rush on, C. Sadegh, D. Sa een, H. Wich e le, S.C. Zhang,
C.N. S endsen, Modeling ALS wi h mo o neu ons de i ed om human
induced plu ipo en s em cells, Na . Neu osci. 19 (2016) 542–553.
[165] M. Luciani, A. G i i, V. Meneghini, Human iPSC-based models o he
de elopmen o he apeu ics a ge ing neu odegene a i e lysosomal s o age
diseases, F on . Mol. Biosci. 7 (2020) 224.
[166] M.A. Lancas e , J.A. Knoblich, O ganogenesis in a dish: modeling de elopmen
and disease using o ganoid echnologies, Science 345 (2014) 1247125.
[167] S.M. Cze niecki, N.M. C uz, J.L. Ha de , R. Menon, J. Annis, E.A. O o, R.E.
Gulie a, L.V. Islas, Y.K. Kim, L.M. T an, T.J. Ma ins, J.W. Pippin, H. Fu, M.
K e zle , S.J. Shankland, J. Himmel a b, R.T. Moon, N. Pa agas, B.S. F eedman,
High- h oughpu sc eening enhances kidney o ganoid di e en ia ion om
human plu ipo en s em cells and enables au oma ed mul idimensional
pheno yping, Cell S em Cell 22 (2018) 929–940 e924.
[168] S.P. Pasca, Assembling human b ain o ganoids, Science 363 (2019) 126–127.
[169] S.N. Bha ia, D.E. Ingbe , Mic o luidic o gans-on-chips, Na . Bio echnol. 32
(2014) 760–772.
[170] M. Sega a, M.R. Abu o, A. Acke -Palme , Blood-B ain Ba ie Dynamics o
Main ain B ain Homeos asis, T ends Neu osci. 44 (2021) 393–405.
[171] C. Lec ux, E. Hamel, The neu o ascula uni in b ain unc ion and disease,
Ac a Physiol. 203 (2011) 47–59.
[172] A.S. Hana y, D. Die ich, G. F icke , A. Lamp ech , Blood-b ain ba ie models:
Ra ionale o selec ion, Ad . D ug Deli . Re . 113859 (2021).
[173] T.E. Pa k, N. Mus a aoglu, A. He land, R. Hasselkus, R. Mannix, E.A. Fi zGe ald,
R. P an il-Baun, A. Wa e s, O. Hen y, M. Benz, H. Sanchez, H.J. McC ea, L.C.
Goumne o a, H.W. Song, S.P. Palecek, E. Shus a, D.E. Ingbe , Hypoxia-
enhanced Blood-B ain Ba ie Chip ecapi ula es human ba ie unc ion
and shu ling o d ugs and an ibodies, Na . Commun. 10 (2019) 2621.
[174] W.M. Pa d idge, Blood-b ain ba ie deli e y o lysosomal s o age diso de s
wi h IgG-lysosomal enzyme usion p o eins, Ad . D ug Deli . Re . 184 (2022)
114234.
[175] R.J. Boado, J.Z. Lu, E.K. Hui, H. Lin, W.M. Pa d idge, Bi- unc ional IgG-
lysosomal enzyme usion p o eins o b ain d ug deli e y, Sci. Rep. 9 (2019)
18632.
[176] A. Bockenho , S. C ame , P. Wol e, S. Knieling, C. Wohlenbe g, V. Gieselmann,
H.J. Galla, U. Ma zne , Compa ison o i e pep ide ec o s o imp o ed b ain
deli e y o he lysosomal enzyme a ylsul a ase A, The Jou nal o neu oscience
: he o icial jou nal o he Socie y o , Neu oscience 34 (2014) 3122–3129.
[177] R.L. Man he, M. Loeck, T. Bhowmick, M. Solomon, S. Mu o, In e wined
mechanisms de ine anspo o an i-ICAM nanoca ie s ac oss he
endo helium and b ain deli e y o a he apeu ic enzyme, J. Con ol Release
324 (2020) 181–193.
[178] E. Hede, C.B. Ch is iansen, C.W. Heegaa d, T. Moos, A. Bu kha , Gene he apy
o he blood-b ain ba ie wi h esul ing p o ein sec e ion as a s a egy o
ea men o Niemann Picks ype C2 disease, J. Neu ochem. 156 (2021) 290–
308.
[179] D. Wang, S.S. El-Amou i, M. Dai, C.Y. Kuan, D.Y. Hui, R.O. B ady, D. Pan,
Enginee ing a lysosomal enzyme wi h a de i a i e o ecep o -binding
domain o apoE enables deli e y ac oss he blood-b ain ba ie , PNAS 110
(2013) 2999–3004.
[180] J.H. G ubb, C. Vogle , B. Le y, N. Gal in, Y. Tan, W.S. Sly, Chemically modi ied
be a-glucu onidase c osses blood-b ain ba ie and clea s neu onal s o age in
mu ine mucopolysaccha idosis VII, PNAS 105 (2008) 2616–2621.
[181] S. Gus a sson, E. Ohlin Sjos om, A. Tje nbe g, J. Janson, U. Wes e ma k, T.
Ande sson, A. Makowe , E. A nelo , G. Ande sson, J. S a eng en, C. Ekholm, S.
S ensson Gelius, In a enous deli e y o a chemically modi ied sul amidase
e icien ly educes hepa an sul a e s o age and b ain pa hology in
mucopolysaccha idosis IIIA mice, Mol. Gene ics Me ab. Rep. 21 (2019)
100510.
[182] K. Ho inouchi, S. E lich, D.P. Pe l, K. Fe linz, C.L. Bisgaie , K. Sandho , R.J.
Desnick, C.L. S ewa , E.H. Schuchman, Acid sphingomyelinase de icien
mice: a model o ypes A and B Niemann-Pick disease, Na . Gene . 10 (1995)
288–293.
[183] H.P. Huang, P.H. Chen, W.L. Hwu, C.Y. Chuang, Y.H. Chien, L. S one, C.L. Chien,
L.T. Li, S.C. Chiang, H.F. Chen, H.N. Ho, C.H. Chen, H.C. Kuo, Human Pompe
disease-induced plu ipo en s em cells o pa hogenesis modeling, d ug
es ing and disease ma ke iden i ica ion, Hum. Mol. Gene . 20 (2011) 4851–
4864.
[184] V.M. Rimkunas, M.J. G aham, R.M. C ooke, L. Liscum, In i o an isense
oligonucleo ide educ ion o NPC1 exp ession as a no el mouse model o
Niemann Pick ype C- associa ed li e disease, Hepa ology 47 (2008) 1504–
1512.
[185] E. Imle , J.S. Pyon, S. Kindelay, M. To und, Y.Q. Zhang, S.S. Chand a, K.E.
Zinsmaie , A D osophila model o neu onal ce oid lipo uscinosis CLN4 e eals
a hype mo phic gain o unc ion mechanism, eLi e, 8 (2019).
[186] H. Kawasaki, T. Suzuki, K. I o, T. Takaha a, N. Go o-Inoue, M. Se ou, K. Saka a,
N. Ishida, Minos-inse ion mu an o he D osophila GBA gene homologue
showed abno mal pheno ypes o climbing abili y, sleep and li e span wi h
accumula ion o hyd oxy-glucoce eb oside, Gene 614 (2017) 49–55.
[187] M.E. Wessels, J.P. Holmes, M. Je ey, M. Jackson, A. Mackin osh, E.H. Kolodny,
B.J. Zeng, C.B. Wang, S.F. Scholes, GM2 gangliosidosis in B i ish Jacob sheep, J.
Comp. Pa hol. 150 (2014) 253–257.
[188] C.H. Vi e, J.H. Bagel, G.P. Swain, M. P ociuk, T.U. Siko a, V.M. S ein, P.
O’Donnell, T. Ruane, S. Wa d, A. C ooks, S. Li, E. Mauldin, S. S ella , M. De
Meulde , M.L. Kao, D.S. O y, C. Da idson, M.T. Vanie , S.U. Walkley,
In acis e nal cyclodex in p e en s ce ebella dys unc ion and Pu kinje cell
dea h in eline Niemann-Pick ype C1 disease, Sci. T ansla ional Med., 7
(2015) 276 a226.
[189] R. K eu ze , M. K eu ze , M.J. P ops ing, A.C. Sewell, T. Leeb, H.Y. Naim, W.
Baumga ne , Insigh s in o pos - ansla ional p ocessing o be a-
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
17
galac osidase in an animal model esembling la e in an ile human G-
gangliosidosis, J. Cell Mol. Med. 12 (2008) 1661–1671.
[190] M.J. Wilke son, D.C. Lewis, S.L. Ma ks, D.J. P ieu , Clinical and mo phologic
ea u es o mucopolysaccha idosis ype II in a dog: na u ally occu ing model
o Hun e synd ome, Ve . Pa hol. 35 (1998) 230–233.
[191] L. Rigon, C. De Filippis, B. Napoli, R. Tomanin, G. O so, Exploi ing he po en ial
o d osophila models in lysosomal s o age diso de s: pa hological
mechanisms and d ug disco e y, Biomedicines 9 (2021).
[192] A. Bachmann, E. Knus , The use o P-elemen ansposons o gene a e
ansgenic lies, Me hods Mol. Biol. 420 (2008) 61–77.
[193] F. Po , H.M. Chen, T. Lee, S.L. Bullock, Op imized CRISPR/Cas ools o e icien
ge mline and soma ic genome enginee ing in D osophila, P oc. Na l. Acad.
Sci. U. S. A., 111 (2014) E2967-2976.
[194] K.G. Hales, C.A. Ko ey, A.M. La acuen e, D.M. Robe s, Gene ics on he ly: a
p ime on he d osophila model sys em, Gene ics 201 (2015) 815–842.
[195] H. B auns ein, M. Papazian, G. Mao , J. Lukas, A. Rol s, M. Ho owi z,
Mis olding o lysosomal alpha-galac osidase a in a ly model and i s
alle ia ion by he pha macological chape one migalas a , In . J. Mol. Sci. 21
(2020).
[196] J.S. Lee, K. Kanai, M. Suzuki, W.S. Kim, H.S. Yoo, Y. Fu, D.K. Kim, B.C. Jung, M.
Choi, K.W. Oh, Y. Li, M. Naka ani, T. Nakaza o, S. Sekimo o, M. Funayama, H.
Yoshino, S.I. Kubo, K. Nishioka, R. Sakai, M. Ueyama, H. Mochizuki, H.J. Lee, S.
P. Sa di, G.M. Halliday, Y. Nagai, P.H. Lee, N. Ha o i, S.J. Lee, A ylsul a ase A, a
gene ic modi ie o Pa kinson’s disease, is an alpha-synuclein chape one,
B ain : A J. Neu ology 142 (2019) 2845–2859.
[197] M.Y. Da is, K. T inh, R.E. Thomas, S. Yu, A.A. Ge manos, B.N. Whi ley, S.P.
Sa di, T.J. Mon ine, L.J. Pallanck, Glucoce eb osidase de iciency in d osophila
esul s in alpha-synuclein-independen p o ein agg ega ion and
neu odegene a ion, PLoS Gene . 12 (2016) e1005944.
[198] S. Ba , M. P asad, R. Da a, Neu omuscula degene a ion and locomo o
de ici in a D osophila model o mucopolysaccha idosis VII is a enua ed by
ea men wi h es e a ol, Disease Models & Mech. 11 (2018).
[199] L. Rigon, N. Kucha owski, F. Ecka d , R. Baue , Modeling
mucopolysaccha idosis ype ii in he ui ly by using he RNA in e e ence
app oach, Li e 10 (2020).
[200] D.L. Webbe , A. Choo, L.J. Hewson, P.J. T im, M.F. Snel, J.J. Hopwood, R.I.
Richa ds, K.M. Hemsley, L.V. O’Kee e, Neu onal-speci ic impai men o
hepa an sul a e deg ada ion in D osophila e eals pa hogenic mechanisms
o Mucopolysaccha idosis ype IIIA, Exp. Neu ol. 303 (2018) 38–47.
[201] G. de Voe , D. Pe e s, P.E. Taschne , Caeno habdi is elegans as a model o
lysosomal s o age diso de s, BBA 1782 (2008) 433–446.
[202] S. Boland, U. Schmid , V. Zago iy, J.L. Sampaio, R.F. F i sche, R. Cze wonka, T.
Lubken, J. Reimann, S. Penko , H.J. Knolke , T.V. Ku zchalia, Phospho yla ed
glycosphingolipids essen ial o choles e ol mobiliza ion in Caeno habdi is
elegans, Na . Chem. Biol. 13 (2017) 647–654.
[203] C. Galles, G.M. P ez, S. Penko , S. Boland, E.O.J. Po a, S.G. Al abe, G.R. Labadie,
U. Schmid , H.J. Knolke , T.V. Ku zchalia, D. de Mendoza, Endocannabinoids in
Caeno habdi is elegans a e essen ial o he mobiliza ion o choles e ol om
in e nal ese es, Sci. Rep. 8 (2018) 6398.
[204] K.P. Bohannon, P.I. Hanson, ESCRT pu s i s humb on he nanoscale: Fixing
iny holes in endolysosomes, Cu . Opin. Cell Biol. 65 (2020) 122–130.
[205] J.M. Huynh, H. Dang, I.A. Munoz-Tucke , M. O’Ke ch, I.T. Liu, S. Pe no, N.
Bhuyan, A. C ain, I. Bo bon, H. Fa es, ESCRT-dependen cell dea h in a
caeno habdi is elegans model o he lysosomal s o age diso de
mucolipidosis ype IV, Gene ics 202 (2016) 619–638.
[206] K. Howe, M.D. Cla k, C.F. To oja, J. To ance, C. Be helo , M. Mu a o, J.E.
Collins, S. Humph ay, K. McLa en, L. Ma hews, S. McLa en, I. Sealy, M.
Caccamo, C. Chu che , C. Sco , J.C. Ba e , R. Koch, G.J. Rauch, S. Whi e, W.
Chow, B. Kilian, L.T. Quin ais, J.A. Gue a-Assuncao, Y. Zhou, Y. Gu, J. Yen, J.H.
Vogel, T. Ey e, S. Redmond, R. Bane jee, J. Chi, B. Fu, E. Langley, S.F. Magui e, G.
K. Lai d, D. Lloyd, E. Kenyon, S. Donaldson, H. Seh a, J. Almeida-King, J.
Lo eland, S. T e anion, M. Jones, M. Quail, D. Willey, A. Hun , J. Bu on, S.
Sims, K. McLay, B. Plumb, J. Da is, C. Clee, K. Oli e , R. Cla k, C. Riddle, D.
Ellio , G. Th eadgold, G. Ha den, D. Wa e, S. Begum, B. Mo imo e, G. Ke y, P.
Hea h, B. Phillimo e, A. T acey, N. Co by, M. Dunn, C. Johnson, J. Wood, S.
Cla k, S. Pelan, G. G i i hs, M. Smi h, R. Gli he o, P. Howden, N. Ba ke , C.
Lloyd, C. S e ens, J. Ha ley, K. Hol , G. Panagio idis, J. Lo ell, H. Beasley, C.
Hende son, D. Go don, K. Auge , D. W igh , J. Collins, C. Raisen, L. Dye , K.
Leung, L. Robe son, K. Amb idge, D. Leongamo nle , S. McGui e, R.
Gilde ho p, C. G i i hs, D. Man h a adi, S. Nichol, G. Ba ke , S. Whi ehead,
M. Kay, J. B own, C. Mu nane, E. G ay, M. Humph ies, N. Sycamo e, D. Ba ke ,
D. Saunde s, J. Wallis, A. Babbage, S. Hammond, M. Mash eghi-Mohammadi,
L. Ba , S. Ma in, P. W ay, A. Elling on, N. Ma hews, M. Ellwood, R.
Woodmansey, G. Cla k, J. Coope , A. T omans, D. G a ham, C. Skuce, R.
Pandian, R. And ews, E. Ha ison, A. Kimbe ley, J. Ga ne , N. Foske , R. Hall, P.
Ga ne , D. Kelly, C. Bi d, S. Palme , I. Geh ing, A. Be ge , C.M. Dooley, Z. E san-
U un, C. Ese , H. Geige , M. Geisle , L. Ka o ki, A. Ki n, J. Konan z, M. Konan z,
M. Obe lande , S. Rudolph-Geige , M. Teucke, C. Lanz, G. Radda z, K.
Osoegawa, B. Zhu, A. Rapp, S. Widaa, C. Lang o d, F. Yang, S.C. Schus e , N.P.
Ca e , J. Ha ow, Z. Ning, J. He e o, S.M. Sea le, A. En igh , R. Geisle , R.H.
Plas e k, C. Lee, M. Wes e ield, P.J. de Jong, L.I. Zon, J.H. Pos le hwai , C.
Nusslein-Volha d, T.J. Hubba d, H. Roes C ollius, J. Roge s, D.L. S emple, The
zeb a ish e e ence genome sequence and i s ela ionship o he human
genome, Na u e 496 (2013) 498–503.
[207] Y.H. Sung, J.M. Kim, H.T. Kim, J. Lee, J. Jeon, Y. Jin, J.H. Choi, Y.H. Ban, S.J. Ha, C.
H. Kim, H.W. Lee, J.S. Kim, Highly e icien gene knockou in mice and
zeb a ish wi h RNA-guided endonucleases, Genome Res. 24 (2014) 125–131.
[208] G.J. Lieschke, P.D. Cu ie, Animal models o human disease: zeb a ish swim
in o iew, Na . Re . Gene . 8 (2007) 353–367.
[209] L.E. Kuil, A. Lopez Ma i, A. Ca e as Masca o, J.C. an den Bosch, P. an den
Be g, H.C. an de Linde, K. Schoonde woe d, G.J.G. Ruij e , T.J. an Ham, Hexb
enzyme de iciency leads o lysosomal abno mali ies in adial glia and
mic oglia in zeb a ish b ain de elopmen , Glia 67 (2019) 1705–1718.
[210] D. Zizioli, M. Gua ien i, C. Tobia, G. Ga iano, G. Bo sani, R. B esciani, R. Ronca,
E. Giacopuzzi, A. P e i, G. Gaudenzi, M. Belle i, E. Di Salle, G. Fab ias, J. Casas,
D. Riba i, E. Mon i, M. P es a, Molecula cloning and knockdown o
galac oce eb osidase in zeb a ish: new insigh s in o he pa hogenesis o
K abbe’s disease, BBA 2014 (1842) 665–675.
[211] M. A ola, C.L. Kuo, L.T. Lelie eld, R.J. Rowland, G.A. an de Ma el, J.D.C.
Codee, R.G. Boo , G.J. Da ies, J. Ae s, H.S. O e klee , Func ionalized
Cyclophelli ols A e Selec i e Glucoce eb osidase Inhibi o s and Induce a
Bona Fide Neu opa hic Gauche Model in Zeb a ish, J. Am. Chem. Soc. 141
(2019) 4214–4218.
[212] E. Mo o, R. Tomanin, A. F iso, N. Modena, N. Tiso, M. Sca pa, F. A gen on, A
no el unc ional ole o idu ona e-2-sul a ase in zeb a ish ea ly de elopmen ,
Ma ix Biology : J. In . Soc. Ma ix Biol. 29 (2010) 43–50.
[213] A.M. Douek, M. Ami i Khabooshan, J. Hen y, S.A. S ama is, F. K eude , G.
Ramm, M.L. Anko, D. Wlodkowic, J. Kaslin, An enginee ed sgsh mu an
zeb a ish ecapi ula es molecula and beha iou al pa hobiology o san ilippo
synd ome A/MPS IIIA, In . J. Mol. Sci. 22 (2021).
[214] K. Okada, R. Takase, Y. Hamaoka, A. Honda, A. Ikeda, Y. Hokazono, Y. Maeda,
O. Hayasaka, T. Ko ani, M. Koma su, K. Shiozaki, Es ablishmen and
cha ac e iza ion o Neu1-knockou zeb a ish and i s abno mal clinical
pheno ypes, Biochem. J. 477 (2020) 2841–2857.
[215] J. Ma suda, O. Suzuki, A. Oshima, A. Ogu a, Y. Noguchi, Y. Yamamo o, T.
Asano, K. Takimo o, K. Sukegawa, Y. Suzuki, M. Naiki, Be a-galac osidase-
de icien mouse as an animal model o GM1-gangliosidosis, Glycoconj. J. 14
(1997) 729–736.
[216] D. Phaneu , N. Wakama su, J.Q. Huang, A. Bo owski, A.C. Pe e son, S.R.
Fo una o, G. Ri e , S.A. Igdou a, C.R. Mo ales, G. Benoi , B.R. Ake man, D.
Lecle c, N. Hanai, J.D. Ma h, J.M. T asle , R.A. G a el, D ama ically di e en
pheno ypes in mouse models o human Tay-Sachs and Sandho diseases,
Hum. Mol. Gene . 5 (1996) 1–14.
[217] T. Ohshima, G.J. Mu ay, W.D. Swaim, G. Longenecke , J.M. Qui k, C.O.
Ca da elli, Y. Sugimo o, I. Pas an, M.M. Go esman, R.O. B ady, A.B. Kulka ni,
alpha-Galac osidase A de icien mice: a model o Fab y disease, PNAS 94
(1997) 2540–2544.
[218] J.J. Mille , K. Aoki, F. Moeh ing, C.A. Mu phy, C.L. O’Ha a, M. Tiemeye , C.L.
S ucky, N.M. Dahms, Neu opa hic pain in a Fab y disease a model, JCI
insigh 3 (2018).
[219] B. Hess, P. Sa ig, D. Ha mann, R. Coenen, R. Lullmann-Rauch, H.H. Goebel, M.
E e s, K. on Figu a, R. D’Hooge, G. Nagels, P. De Deyn, C. Pe e s, V.
Gieselmann, Pheno ype o a ylsul a ase A-de icien mice: ela ionship o
human me ach oma ic leukodys ophy, PNAS 93 (1996) 14821–14826.
[220] L.W. Duchen, E.M. Eiche , J.M. Jacobs, F. Sca a illi, F. Teixei a, He edi a y
leucodys ophy in he mouse: he new mu an wi che , B ain : A J.
Neu ology 103 (1980) 695–710.
[221] I. Jones, X. He, F. Ka ouzian, P.I. Da och, E.H. Schuchman, Cha ac e iza ion o
common SMPD1 mu a ions causing ypes A and B Niemann-Pick disease and
gene a ion o mu a ion-speci ic mouse models, Mol. Gene . Me ab. 95 (2008)
152–162.
[222] G.B. Sinclai , G. Je on, K.E. Colobong, D.R. Randall, F.Y. Choy, L.A. Cla ke,
Gene a ion o a condi ional knockou o mu ine glucoce eb osidase: u ili y
o he s udy o Gauche disease, Mol. Gene . Me ab. 90 (2007) 148–156.
[223] J. Ma shall, K.A. McEache n, J.A. Ky os, J.B. Nie upski, T. Budzinski, R.J. Ziegle ,
N.S. Yew, J. Sulli an, A. Sca ia, N. an Rooijen, J.A. Ba ange , S.H. Cheng,
Demons a ion o easibili y o in i o gene he apy o Gauche disease using
a chemically induced mouse model, Mol. The apy : The J. Am. Soc. Gene
The apy 6 (2002) 179–189.
[224] J. Muenze , J.C. Lamsa, A. Ga cia, J. Dacos a, J. Ga cia, D.A. T eco, Enzyme
eplacemen he apy in mucopolysaccha idosis ype II (Hun e synd ome): a
p elimina y epo , Ac a Paedia . 91 (2002) 98–99.
[225] L.A. Cla ke, C.S. Russell, S. Pownall, C.L. Wa ing on, A. Bo owski, J.E.
Dimmick, J. Toone, F.R. Ji ik, Mu ine mucopolysaccha idosis ype I: a ge ed
dis up ion o he mu ine alpha-L-idu onidase gene, Hum. Mol. Gene . 6
(1997) 503–511.
[226] M. Bhaumik, V.J. Mulle , T. Rozaklis, L. Johnson, K. Dob enis, R. Bha acha yya,
S. Wu zelmann, P. Finamo e, J.J. Hopwood, S.U. Walkley, P. S anley, A mouse
model o mucopolysaccha idosis ype III A, San ilippo synd ome,
Glycobiology 9 (1999) 1389–1396.
[227] H.H. Li, W.H. Yu, N. Rozengu , H.Z. Zhao, K.M. Lyons, S. Anagnos a as, M.S.
Fanselow, K. Suzuki, M.T. Vanie , E.F. Neu eld, Mouse model o San ilippo
synd ome ype B p oduced by a ge ed dis up ion o he gene encoding
alpha-N-ace ylglucosaminidase, PNAS 96 (1999) 14505–14510.
[228] E.H. Bi kenmeie , M.T. Da isson, W.G. Beame , R.E. Ganschow, C.A. Vogle , B.
Gwynn, K.A. Ly o d, L.M. Mal ais, C.J. Waw zyniak, Mu ine
mucopolysaccha idosis ype VII. Cha ac e iza ion o a mouse wi h be a-
glucu onidase de iciency, J. Clin. In es . 83 (1989) 1258–1266.
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
18
[229] S. Toma su, K.O. O ii, C. Vogle , J. Nakayama, B. Le y, J.H. G ubb, M.A.
Gu ie ez, S. Shim, S. Yamaguchi, T. Nishioka, A.M. Mon ano, A. Noguchi, T.
O ii, N. Kondo, W.S. Sly, Mouse model o N-ace ylgalac osamine-6-sul a e
sul a ase de iciency (Galns-/-) p oduced by a ge ed dis up ion o he gene
de ec i e in Mo quio A disease, Hum. Mol. Gene . 12 (2003) 3349–3358.
[230] M.J. P zybilla, L. Ou, A.F. Taba an, X. Jiang, R. Sidhu, P.J. Kell, D.S. O y, M.G.
O’Sulli an, C.B. Whi ley, Comp ehensi e beha io al and biochemical
ou comes o no el mu ine models o GM1-gangliosidosis and Mo quio
synd ome ype B, Mol. Gene . Me ab. 126 (2019) 139–150.
[231] M. E e s, P. Sa ig, P. Schmid , A. Ha ne , D.B. McLoghlin, W. Schmahl, B. Hess,
K. on Figu a, C. Pe e s, Ta ge ed dis up ion o he a ylsul a ase B gene esul s
in mice esembling he pheno ype o mucopolysaccha idosis VI, PNAS 93
(1996) 8214–8219.
[232] M. Yoshida, M. Tachibana, E. Kobayashi, H. Ikadai, T. Kunieda, The locus
esponsible o mucopolysaccha idosis VI (Ma o eaux-Lamy synd ome) is
loca ed on a ch omosome 2, Genomics 20 (1994) 145–146.
[233] R.J. Ro ie , E. Bon en, A. d’Azzo, A poin mu a ion in he neu-1 locus causes
he neu aminidase de ec in he SM/J mouse, Hum. Mol. Gene . 7 (1998) 313–
321.
[234] X.Y. Zhou, H. Mo eau, R. Ro ie , D. Da is, E. Bon en, N. Gillemans, D. Wenge ,
F.G. G os eld, P. Dohe y, K. Suzuki, G.C. G os eld, A. d’Azzo, Mouse model o
he lysosomal diso de galac osialidosis and co ec ion o he pheno ype
wi h o e exp essing e y h oid p ecu so cells, Genes De . 9 (1995) 2623–
2634.
[235] S. S inchi, R. Lullmann-Rauch, D. Ha mann, R. Coenen, T. Becca i, A.
O lacchio, K. on Figu a, P. Sa ig, Ta ge ed dis up ion o he lysosomal
alpha-mannosidase gene esul s in mice esembling a mild o m o human
alpha-mannosidosis, Hum. Mol. Gene . 8 (1999) 1365–1372.
[236] M. Zhu, K.L. Lo ell, J.S. Pa e son, T.L. Saunde s, E.D. Hughes, K.H. F ide ici,
Be a-mannosidosis mice: a model o he human lysosomal s o age disease,
Hum. Mol. Gene . 15 (2006) 493–500.
[237] H. Wol , M. Damme, S. S ooban s, R. D’Hooge, H.C. Beck, I. He mans-
Bo gmeye , R. Lullmann-Rauch, T. Die ks, T. Lubke, A mouse model o
ucosidosis ecapi ula es s o age pa hology and neu ological ea u es o he
milde o m o he human disease, Disease Models & Mech. 9 (2016) 1015–
1028.
[238] V. Kaa inen, I. Mononen, J.W. Voncken, T. No onkoski, I. Gonzalez-Gomez, N.
Heis e kamp, J. G o en, A mouse model o he human lysosomal disease
aspa ylglycosaminu ia, Na . Med. 2 (1996) 1375–1378.
[239] S.K. Lo us, J.A. Mo is, E.D. Ca s ea, J.Z. Gu, C. Cummings, A. B own, J. Ellison,
K. Ohno, M.A. Rosen eld, D.A. Tagle, P.G. Pen che , W.J. Pa an, Mu ine model
o Niemann-Pick C disease: mu a ion in a choles e ol homeos asis gene,
Science 277 (1997) 232–235.
[240] R.A. Maue, R.W. Bu gess, B. Wang, C.M. Wooley, K.L. Sebu n, M.T. Vanie , M.A.
Roge s, C.C. Chang, T.Y. Chang, B.T. Ha is, D.J. G abe , C.A. Pena i, D.M.
Po e , B.S. Szwe gold, L.P. Hende son, J.W. To enhagen, T.P. T oua d, I.A.
Bo bon, R.P. E ickson, A no el mouse model o Niemann-Pick ype C disease
ca ying a D1005G-Npc1 mu a ion compa able o commonly obse ed
human mu a ions, Hum. Mol. Gene . 21 (2012) 730–750.
[241] M. P aggas is, B. To elli, J. Zhang, H. Fujiwa a, R. Sidhu, A. Chacko, Z. Chen, C.
Chung, A.P. Liebe man, J. Siko a, C. Da idson, S.U. Walkley, N.H. Pipalia, F.R.
Max ield, J.E. Scha e , D.S. O y, A mu ine Niemann-Pick C1 I1061T knock-in
model ecapi ula es he pa hological ea u es o he mos p e alen human
disease allele, The Jou nal o neu oscience : he o icial jou nal o he Socie y
o , Neu oscience 35 (2015) 8091–8106.
[242] Y. Tanaka, G. Guhde, A. Su e , E.L. Eskelinen, D. Ha mann, R. Lullmann-Rauch,
P.M. Janssen, J. Blanz, K. on Figu a, P. Sa ig, Accumula ion o au ophagic
acuoles and ca diomyopa hy in LAMP-2-de icien mice, Na u e 406 (2000)
902–906.
[243] J.P. Mo gan, H. Magee, A. Wong, T. Nelson, B. Koch, J.D. Coope , J.M. Weime , A
mu ine model o a ian la e in an ile ce oid lipo uscinosis ecapi ula es
beha io al and pa hological pheno ypes o human disease, PLoS ONE 8
(2013) e78694.
[244] A. Ba oll, A. Toledano-Za agoza, J. Casas, M. Guzman, E.H. Schuchman, M.D.
Ledesma, Inhibi ion o a y acid amide hyd olase p e en s pa hology in
neu o isce al acid sphingomyelinase de iciency by escuing de ec i e
endocannabinoid signaling, EMBO Mol. Med., (2020) e11776.
[245] H. Lowe, N. Toyang, B. S eele, J. B yan , W. Ngwa, The endocannabinoid
sys em: a po en ial a ge o he ea men o a ious diseases, In . J. Mol.
Sci. 22 (2021).
[246] K.K. Kumme , T. Kalpachidou, M. Mi ic, M. Langeslag, M. K ess, Al e ed gene
exp ession in p e on al co ex o a ab y disease mouse model, F on . Mol.
Neu osci. 11 (2018) 201.
[247] A.R. Ga cia, J. Pan, J.C. Lamsa, J. Muenze , The cha ac e iza ion o a mu ine
model o mucopolysaccha idosis II, Hun e synd ome, J. Inhe i . Me ab. Dis.
30 (2007) 924–934.
[248] N. de Gees , E. Bon en, L. Mann, J. de Sousa-Hi zle , C. Hahn, A. d’Azzo,
Sys emic and neu ologic abno mali ies dis inguish he lysosomal diso de s
sialidosis and galac osialidosis in mice, Hum. Mol. Gene . 11 (2002) 1455–
1464.
[249] G.B. Baskin, M. Ra e ee, B.B. Da ison, K.P. Falkens ein, M.R. Cla ke, J.D.
England, M.T. Vanie , P. Luzi, M.A. Ra i, D.A. Wenge , Gene ic
galac oce eb osidase de iciency, globoid cell leukodys ophy, K abbe
disease) in hesus monkeys (Macaca mula a, Lab. Anim. Sci. 48 (1998)
476–482.
[250] D. Golebiowski, I.M.J. an de Bom, C.S. Kwon, A.D. Mille , K. Pe osky, A.M.
B adbu y, S. Mai land, A.L. Kuhn, N. Bishop, E. Cu an, N. Sil a, D. GuhaSa ka ,
S.V. Wes mo eland, D.R. Ma in, M.J. Gounis, W.F. Asaad, M. Sena-Es e es,
Di ec in ac anial injec ion o AAV h8 encoding monkey be a-n-
ace ylhexosaminidase causes neu o oxici y in he p ima e b ain, Hum.
Gene The . 28 (2017) 510–522.
[251] F. DeRosa, L. Smi h, Y. Shen, Y. Huang, J. Pan, H. Xie, B. Yahalom, M.W.
Hea lein, Imp o ed e icacy in a ab y disease model using a sys emic mRNA
li e depo sys em as compa ed o enzyme eplacemen he apy, Mol.
The apy : The J. Am. Soc. Gene The apy 27 (2019) 878–889.
[252] J.B. Rosenbe g, D. Sondhi, D.G. Rubin, S. Mone e, A. Chen, S. C am, B.P. De, S.
M. Kaminsky, C. Se in, P. Aubou g, R.G. C ys al, Compa a i e e icacy and
sa e y o mul iple ou es o di ec CNS adminis a ion o adeno-associa ed
i us gene ans e ec o se o ype h.10 exp essing he human a ylsul a ase
A cDNA o nonhuman p ima es, Human gene he apy, Clin. De . 25 (2014)
164–177.
Á. Gaudioso, T.P. Sil a and Ma í. Dolo es Ledesma Ad anced D ug Deli e y Re iews 190 (2022) 114532
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