Multinodular and Vacuolating Neuronal Tumour

doi:10.1111/bpa.12555 between this versionandtheVersion ofRecord.Pleasecite this articleasan‘Accepted Article’, the copyediting,typesetting, paginationandproofreadingprocess,which mayleadtodifferences This articlehasbeenaccepted forpublicationandundergonefullpeerreview buthasnotbeenthrough Tel:++44 20 43884233 Email:[email protected] UK WC1N3BG London SquareQueen UCL Instituteof Neurology Departmentof Neuropathology MariaThom Correspondingauthor: 9 8 7 6 Neurosurgery,Queen Square, London 3BG, UK WC1N 5 Institute ofNeurology, QueenSquare 3BG, LondonWC1N 4 3 2 London. UK. WCN1BG, Neurologyand National the Hospital Neurology for Neurosurgery, and Queen Square, 1 Jacques Miserocchi Jäger Maria Thom DYSPLASIANEOPLASIA?OR MULTINODULARANDVACUOLATING NEURONAL TUMOURSEPILEPSYIN Stichting EpilepsieStichting InstellingenNederland (SEIN), TheNetherlands NeuropathologyDepartment, GreatOrmond Street Hospital Departmentof Anatomic Pathology, Hospital Pedro Hispano,Matosinhos, Portugal NeurosurgeryDepartment, Brighton and Sussex UniversityHospitals,Brighton, UK HorsleyVictor Department of Neurosurgery, National Hospital Neurology for and NeuroradiologicalAcademic Unit,Department Repairof Brain and Rehabilitation, UCL PathologyUniversityUnit, Padua of Medical School Departmentsof Clinical and Experimental Epilepsy Departmentof (Neuro)Pathology, Academic Medical Center, Amsterdam, The Netherlands

4 , Cheryl Reeves 8 , Eleonora Aronica 5 , Lewis, Thorne 1

Accepted Article Liu ,Joan 1 , Alyma, Somani 1 This article isprotected by copyright. All rights reserved. This article isprotected by copyright. All rights reserved. , Anika Bongaarts, 5 2 , Fay Newman ,9 . 1 , Shu An , Shu 6 , Sorin Bucur, Sorin 2 , R oy 1 , Derek Marsdon .J. Reinten .J. Neuropathology,and UCL Instituteof , Padova,IT. 6 , Mrinalini Honavar 2 , , Beatrice Paradiso London 1 , Andrew McEvoy , Andrew , UK , 7 7 , To 1 ,3 : , Hans Rolf m 5 , Anna , Anna neoplasmwith origin afrom geneticdata could indicate detectedin oligodendrogliallineage markers (OLIG2,SMI94) developmenta (n surgeryof was incases seriesthis identify mutationscommon to epilepsy l changes differentiation,interneuronal populations, mTOR pathwayactivatio and a grade ep epilepsyasso ten and fifteen includedin the Multinodularand vacuolating Abstract eurofilament/SMI32,

casesand compare a neoplastic a

broad i lack of reportsin literature the . ilepsyassociated tumours(LE

Next generationsequenci 45

the the mmunohistochemistry ciatedmalformations including focalcortical dysplasia type(FCDII)II and low any years. lly recently mTOR

regulated proteins (OTX1, versus malformative biologyhas

Accepted pr Article

presentedwith seizures ogressive change The vacuolatedcells of lesion the expressed mature neuronalmarkers

thatMVN progenitor

pathologicaland genetic features betterto characterised

neuronal tumour todate. Theyare

ngwas performed panel T s ,

align AT neuro BRAF, esionsincluding FCDII and LEAT on serialon pre

). was

, classification TBR1, s Clin ) and werelocatedin - more withmalformative a lesion athan tr . P glial

(MVNT) F a typicallyassociated with onset late epilepsy icaland neuroradiology data werereviewed GF pplied exploreto neuronal and g rominentlabelling of

was observed been celltype showing SOX2,MAP1b, CD34, GFAP R1 orMYB - operative

fortarget

isa new pattern

questioned

of tumoursof the CNS . MRIand ed multi Clinical, pathological .

the temporallobe No n andneurodegenerative . presenta We series of

aberrantmaturation. mutations thelesion

- of a mean ageat geneanalysis .

Allth of neuronaltumour alcells were . . There are δ lial ) esurgical . . There and

and

ue to to

for for

CASE SELECTION CASE Methods tumour better maturation, MVN In studythis CNSneoplasms that characterizationfurther i commentedthat lesthese pattern entitythis isnow included the in have beentermed considered (FCD matterby Pathology inpatient a with onset late epilepsy 20 In masseffect, assuch tuberous sclerosis. characterizedby ab or ha gangliogliom progressive developmentalanomalies I Introduction ndolent ma those those T 07 ) r

,mild malformation tomatousor

explore

(includingthe case our in initial report , of gangliocytoma in the contextof long

cortical w assoc nodules ofvacuolat e reporteda unique case ofa examination ofan MRI visible lesion

in addition

geneticabnormalities, , neurologyor MRI changes

aand dysembryoplastic neuroepithelialtumours(DNT) we review the

itsdelineation between corticalmalformation iatedwith mutationsin the

lesions ( 33

Accepted Article

‘ d Multinodularand normalcorticalarchitecture with ) evelopmental .

to a neuronal tumour

provoking focal

ions‘may even malformativebe andnature’ in ofcortical development, nodular heterotopia were and utwith ‘uncertain class clinical - s neededs to fullyunderstand their nosological placeamong term focalepilepsy. ed ganglioncells

lowgrade tumour current

‘overgrowth’

and and which posed vacuolating neuronaltumour

potential

arecharacteristic of pathological refra

2016 cl WHO ‘ diffuse gangliocytoma PI3K .

ctory

( Recently 45 – ;

disclosed AKT malformationsas lesions s epileptogenic mechanisms )

) .

such a such . e We assignment epilepsyoftenbridge a

excessive cellsize andtumour findings s long assificationof CNS tumou ignallingpathway

reportso , particularly , the mTORopathies

diagnosticconundrum diffuse plorethe history

oran WHO grade WHO in aseries ’ ( ’ 34 f involving involvemento

histologically s epilepsy f ofseizures and ) ocalcortic

. ( ce ( M I 57 ndeed,i llulardifferentiation and VNT) ) . often ( I 21

itisacknowledged

the gapbetween tumours - Ho associated ’ ) ( inaim an , diagnose 7 aldysplasia

t w are temporallobe

, has been cases ever similarlesions f whitef the 17

rsas a ( 45

, a ,such 24 , - ) lackof

like . many of ) to s

and

focal focal

(

)

instructions. TissueDNA Isolation (MOkit Carlsbad,BIO, CA, USA)according tothe manufacturer’s dissectedmicro 10from μm tissue sections.DNA was extractedusing the BiOsticFFPE mortem samples Nextgeneration s SEQUENCING GENERATION NEXT ou carried perf (pS6 neurode (c cellmarkers corticallayer specific markers material common glioma mutations (IDH1,ATRX and BRAF V600E) oligodendrogliallineage and myelination neurofilament paraffinfixed, block anda A RoutineH IMMUNOHISTOCHEMISTRY for 32 with glioneuronal tumour Devel research forthis study ( MedicalCentre TissueBank at and malformations Sevenc London utostainer albindin )

im ) ormed( , and , mild malformationof cortical development typeII ,

munohistochemistry opmentalBiology Resource HDBR Ser240/244 andpS6 ases ge , case4

available .

(case7 notsequenced UK , FCD & t t manuallyusing standard protocols We n calretinin,parvalbumin, NPY)

Eand BondMax Auto erative mar (Leica,Milto panelof ) and ) NGS (CD34,Reelin and of - s

embeddedbrain sections also ,synaptophysin, MAP2)

type

failedsequencing due lowto quality DNAof UCL ( M Amsterdam

equencing allsurgical patients VNT LFB

was performedusing a Accepted Article HospitalPedro Hispano,

a more extensive immunohistochemistry

inthe include II casesII

. (WHO grade I)

(Supplemental Table 1 fordetails) or genetics d wereperformed on

, epartment mated Immunostainer ,

,Netherlands from epilepsyfrom surgical patients (p62,AT8, APP, mitochondria) PAX6, f Ser235/236) (NGS)of MVN etalde

(TBR1, TBR2,OTX1 three

due lackto of availability of frozen tissue

, SOX2,Nestin, DCX

velopmental controls(from casesof MVNT temporallobe cortex ned fro consented for of , .

astroglialdifferentiation Fromall , toexplore N , (Olig2, chloride ), customised UCL . europathology

Either a T ( Great Ormond StreetHospital children, for mpathology review all of glio markers Matosinhos wasout carried in eightof thenine

), sectionsof as previouslydetailed S conventional case

(Leica, MI94/ co utomat : , N200, , MAP1B)

neuronaldifferentiation useof

- wereincluded (

transporters Table 1)was s (increasedwhite matterneurones Ion AmpliSeq™ Ion Neurology Panel

, PDGFRβ , myelin basicprotein w ,

and EpilepsySoci e MiltonKeynes, Portugal adjacent hippocampalto scle all ed immunohistochemistred

tissue then then

) , . and panelwas appliedincluding

as comparative control groups cases In Tumourtissue was manually g

angliogliomas sevencases with sufficient MRC

(GFAPand GFAPδ) selected a mTORpathway , , clinicaland MRI data

) ) (NKCC1,NKCC2) . ,interneuronal subsets

from applied using theLeica ,

The - ( devel Wellcome TrustHumanWellcome 20

) reise theAcademic UK)

stud -

(Table 1) neuronaltum and opmental/

representative e on

(NeuN, (MBP) tyBrain and ) or it) was /lowgrade ies.

thepost 5 µm formalin thicalapproval

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surgical y ) .

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and was stem , , ours rosis

in ( 6

, - informationavailable (Table 2) Six the ten of CLINICAL FEATURES ( diagnosed condition t O epilepsy theOf Results ( cases ( d updatedpost Thecli CLINICAL DATAANDMRI compar genes genewere developed identifyto most mutations SeqNext coverage of1500X per amplicon was established werebarcodedusing theIonXpress Barcode Adapterseach for 318 Ion V2 Chip.A mean was 20to up ng, which was 318 V2Chip andPersonal Genome Machinewere usedas sequencing platform foremulsion PCR and template preparation. the AmpliSeq Ion Library Kit2.0. PGMThe Ion Hi PTCH1,PTEN, SMARCA4, SMARCB1, SMO,SUFU, TP53.Libraries were preparedusing IDH1, KDM6A,IDH2, mTOR, MYB, MYBL1, NPRL2, NPRL3, PIK3CA, PIK3R1, PIK3R2, CTNNB1,CIC, DDX3X, amplification (ThermoFisherScientific, Massachusetts, Waltham, USA) for targeted multi wo all H i fferent ne RJ) inRJ) additionto

reported since2014) further

surgicalcase . . G nical notes ten Case

edto 22 samples , MR

ene (JSIMedisys, Ettenheim, Germany). Of and a further case (Case

cases

assubcortical mattergrey analysed surgicalcases I . panel This

- 1,2

sequence duplications cases operative informatio s ,5,6,8

with 2 and3

Accepted f Article was rom

po , were females. thusthere is M st and

eachcase were

T, followinghistopathological assessment reports

measured bymeasured the was

nine protocols.All

FCDtype II

recogniz with

notdetec initial s identified

n of the followinggenes;AKT1, AKT3, ATRX, BRAF, CDK6, r weresurgical resections

in theremainder) eport .MRI The The mean ageat seizure onset

arisk of undetected mutationsin tumor suppressor a mean a ly

in heterotopia

ed as M ed

diagnosedas findingsof the reviewedforpresenting neurological edas an unusual

ted had been carriedout in

and8 glioneuronaltumours atpost available pre genes , T

ge atge surgery of 45 andt Qubit3.0 Fluorometer. Up tospecimens 20 he PGM Ion Hi VNT

note,the selected sequencing panel was , - (Case4) -

QKitand Ion Chef Instrument were used mortem studied he and data the anydisease .

were reviewed

a sequence

gangliocytomaand a demyelinating - operative .However, all not exons each of

. The remaining six gangliocyt asincidentalan finding carriedout for treatmentof - Q sequencing Q with Kit theIon progression/recurrence was analyzedusing JSI

analysisMVNof

years different ;t MRI was

bya he oma .

(range6 67 to

PMcase scans ofscans 29.7 neuroradiologist

centres (Case1) -

gene years symptoms

surgical casessurgical

s T were T was surgical . DNA . in

using

(Table 2

based ( 45

years) ) .

and

put on ) . .

component, reduced on extentof the white mattercomponent diffuse subcorticalwhite ma corticallayers. nodules His subcorticalwhite matte (Figure 2 lo translucency Ma NEUROPATHOLOGYFINDINGS showedprogressive changes diagnosisor, less likelycorticaldysplasia, based o periodsof up to period,in onecase with (Figure 1a,b). There were investigated associatedwith these lesions extentof th contourwas apparentin butsome without clea involvement of the FLAIRsequenc abnormality S NEUROIMAGINGFEATURES progression becomedid not seizure fourcases were occipitallobe (hippocampus,amygdal In five The alientMRI features be whitebe matter

croscopic myelin s tologicalexamination single

surgicalcases ,

e). predominan post eabnormality varied between tained sections (LFB/CVor SMI94/MPB

.Clear involvement I in on MRI,on even in caseswith partial resections

nthe post . abnormalities orcavitation

C thetemporal lobe, Coalescingor abutting nodular islands (Figure2i) as well as areas of more - es linical mortem eight recentlyoperated (Figure2a) Accepted Article,

adjacent in , , the lesions volving cortex the tly commonto all cases included synaptophysin

years ( outcomes - r - mortemspecimen a orparahippocampal gyrus) located free case of sixinterval MRI studies (

ofthe no progressiveno changesserialon images in the pre

the the noted tterinvolvement (Figure 2c

white matter Case post

. . Therewas enhancement no or restricted or was during the right

in the

ofthe hip with subcortical wereavailable - typically in primarilyinvolved the temporal lobe, operatively,

a 62 1 with occipitallobe , Figure , 1a) fixed, (Figure 2f) plaque atlow power subcortical

andstraddling the subcortical regionwith moreclear less than yearold

post

insome cases (Figure 1).A vague, corrugated cases pocampus was apparent ,nodules of grey matterwere noted in the hypo resected

- white matter like nodulesislands or of -

operative andcase no r nodularity r cysticor change . -

,there was no . Low grade glioma intense toup years14 post male In regionsbearing C two only white matteror ‘sitting’ in thedeeper n imaging ase

) was bestappreciated by reduced lobectomy poorlydefined regionsof

(Figure2b, c, ; years . withouta clinicaldiagnosis of epilepsy. the post the ) were )

period (Table 2). on T ,Figure 1c,d in

(Table 2)

ha the of

1 features

observed follow - or minimal with hyperi specimensincluded mortemlesio gyralcores documented

d in

. - was consideredthe likely cortical - , i operatively

) .None of the lesions up. up. Onlyone patient C ) grey discolo and and ases . and corticalfor the

diffusion The architecture

ntens masseffect

over ofthe temporal . A mesialstructure nodules, n involved 1,2, 8and 9 lesion s - lthoughthe ignal operative ityon T2and , however , review

where ur

focal ation the the

myelin the the

and s

CORTICAL LAYER DIFFERENTIATION MARKERS DIFFERENTIATION LAYER CORTICAL noted ganglioglioma showedweak corticalto the s weaker proportionof but neurofilamentcocktail and O expression in Immunolabellingusing the m DIFFERENTIATION NEURONAL a temporallobe bodies (EGBs) cytology gyrus white matterunderlyi In were M (Figure impressionin some predominantVC arrangements was minimalclustering and no with nucleoli Thenodules werecomposed of laminarcyto ynaptophysin

the itotic activity tumourwith hyrbrid features VC five VC

r n , in , present not

white matter compared VC.to the cases

were mainlynegative eurofilamentantibodies

2m in VC with

mor keepingwith previousreports eosinophilic ) for

- althoughin other ,

ety frequent architectureand myelo thesmall multipolar cells in - t

compared to

white matterborder

cytoplasmic

he component,strong alongside and

n component expression (Figure on , pical for gangliogliomawith

and Accepted Articleinvolvementof s

- maller and significantple phosphorylated

nodules was of ng the subiculum ng (Figure2d

prominentperivascu cytopl

cytoplasm

and ) nodule in labell hadthe N200 labelled

other asmic

. atureneuronal marker thenodules

cases( . MA -

orientation like Hippocampa (Table 1), ing in alling cases large cellswith

the om werenoted o . er P and r

verlyingcortical neurones. Despite an vac atlow magnification In 2 highlighted 2 highlighted VC typical glia

neurofilament - diminished cellularity synaptophysin labellingof o architecture of cortex the hippocampus C C r u phismwas lacking l

ase ase s ofs spindled astroglial cells are

o ( cellswere 7 lation or peri

nodules , (Figure 3e), only

includingphosphorylated neurofilament appearances ofMVN r of lar 24

7 8, bi

lsclerosis dysmorphic gangliodcells,eosinophilic granular ) vacuolated in o

) - infiltrates the regions of

nucleate cells occasional but

(Figure 3d) overallgangliocytic morphology ne case

), CA1), andas well as . mesialtemporal component

(

I we confirmed intermingled, SMI32 was present n ,

and NeuN Cases -

cellular impartinga

was (Figure were

diffuse cells . Rosenthal . fibres or

dendrites

(Figure ) comparedto adjacent axonstraversing

, in VCin .In

also noted was 3and 4, were notedin

vacuolation (VC)

was C atypicalganglion cells was 2 , T (Figure T

hypercellularitywere noted

f ase weak but with islands of

consistent, ); 2l

‘moth present

(Figure aproportion of VC ) within otherwise undisturbed.

allcases

. without

even though 8with the Inaddition to the four

ornegative ( inthree cases

parahippocampal - eaten’appearance

MAP2

overall

nodules but 2h

(

cases. (Figure2

Figure2j 2h, throughnodules a

sate ) strong

(Figure

focal focal ; ; suggestedthis calcification showed 3c VC labelling ,

white matter

llitosis. The reductionof prominent )

the the (SMI31) a

k Figure 3 in ) nd and )

. 3b .

. There

C the the

ells ) . was , . a ) cells werealso wereseen not VCwere la NEURODEVELOPMENTAL rims cytoplasmic reduction negative awayvesselsfrom NG2/oligodendroglialprogenitor surgical positive (Figure 3j) inset small localisingto glial bottom lesion the GFAP GLIAL overlying the cortex. 3h intense cytoplasmicl matter appeared was negative post case dysplastic labellingof while TBR Applicationcortical of layer and )

in -

1 tumour ) . of mortem

in the viciniin the LINEAGE MARKERS LINEAGE . in ;a proportion of VC in multipolar I

additionto n In keepingwith previousreports accentuatedvariable proportionsof small, multipolar as emptyvacuoles inset) . tense c other other cases casesor

NG2 andPDGFRα with

of wellas a mainly ; typically neuronswere negative

rgelynegative for reelin neuronal aproportion of the

specific component was confirmed membrano m thenodules and

case

PDGF withmarker this

In yelinated infibres lesion the with ytoplasmiclabelling ofVC for OTX1 was noted

glial ty of thetyof nodules C

labelling ofpyramidal cells of normal morphologyin l negative with PAX6 post delineating the

Accepted Article

ase (Figure 3g) in the mergingwith denser a gliosisin the as wellas , R

marker to

abellingof the VC cells cellsin VC wereweak

β AND INTERNEURONAL INTERNEURONAL AND - mortem 8 with a .

us also highlighted MBP

in all or granularlabelling four of eightfour immuno proximityto VC and sometimes enveloping them diffusecomponents low at magnification

in (SMI94)showed str

in ; distinguish smaller glialcells associatedwith the tissue

ganglioglioma

cortical in comparison neuronal

adjacenttissue

cells keepingwith , extentof the MVN (Figure4a) with TBR1. although ,

and or negativeor markers .

( which However 18 .

layers andtheir dendritic likeprocesses ( .

7 cellswith the morphologyo

GFAP

MNVN ,

, linage MARKERS VC from 55 24 asdescribed above (Figure2c,i) theywere showednuclear labelling of a proportion . ) . ) did n , T component,a p C

theover In In VC small δ of VCwas noted

reviousreports here was n

. onsistent

PDGFRβ,which labels were markersrevealed

showedstrong showed Theo C iking a iking

ot showot any consistent showedstrong nuclearlabelling for OLIG2 ase normalinterstitial neurones T . multipolarcells

noted With With present lyingcortex verlyingcortexshowed 8 with the ganglioglioma surrounding bnormalities,correlating with overall

restrictedexpression MVNin c o labell ytoplasmic or membranous MAP

(Figure astr moreprominent GFAP

in theoverlying corte

(

in allc

oglial 20 nucle 1b for MBP for (Figure and3l) the ing of ing

) variab , andnormal white matter .OTX1

f Caj f we 3k) white matter

lesion within arpositivity

cells ases ayer (Figure 3i)

also observed both both ; VC al le labellingVC of the large VC in allcases (Figure labelling , -

and

Re nodulesin the we apart from the therefore .In addition, for

II,III pe

re OLIG2 re tzius cells (Figure

TBR1 in thewhite TBR ricytes and processes

component

(Figure3i,

(Figure 3f) andV labelling of of the - x 2 positive .

nuclear small

in any

VC 3i were T, T, -

, , top

for in ,

positive (Supplementarycase Figure 1a) In OF MVN COMPARISON BRAF V600Emutation cells l Ahigh proportion of VC showed antibodies gang cells cytoplasm minimallabelling matter and changes comparedto the cortical virtually noneIn of the MTOR NEURODEGENERATIVE, wasdis less 4 impressionan for reduced morphology ( and white matter (Figure 4f) processesin some nodules (Figure associated werepositive extensive labelling(in multipolarcells and serpiginous islands of lesion the at low magnification labe abellingwhich was C f ) ase 3) ase gangliogliomacontrols .

KCC1 lling of VClling of anti

lioglioma ( overlyingin the cortex

overall<5% neurones nodulesof the M - negative. labelling .

which showed mitochondrialantibodies VC

showed

was noted of showed within thelesion t and axonalnetworks inct

smal showed (Figure4d,e surgical

componentof in werepresent in . . . VC . were generallynegative with these

adjacent

( ofVC

M With mTOR pathwayWith antibodies SupplementaryFigure 1b)but more

Acceptedl Article ) In strong . glial VN a T TO virtuallyinabsent VC, In allIn MVN

the normal in VCsome cytoplasmic la C T s werenegative caseswere tangles noted in VC

was obs ases CONTROLS PVexpression likecells associated with thelesional nodules

cytoplasmic labell ),

cortex(Figure 4i PATHWAY PATHWAY Tbr1 numbers compared to adjacenttissue

. T (Figure4l insets) ; bothmarkers showed

Case morphologyand distribut , 7 SOX2 - Nestin

mortem in fact , T

8) nuclear allcases ervedallacross cases

;a nuclearlabel cases,

, GROUPS (Figure4c) were

(Figure4k) in contrastto 8.However, s

variable proportionalso showed cytoplasmic AND CELL CYCLE MARKERS CYCLE CELL AND 4 was seen belling , also

e inset ; werestrikingly

case .

in also

label immunohistochemistry for ,

highlighted VC ranging veryfrom occasional cell but )

with APP ing of ing VC with suggestinglate acquisition of degenerative intermingled ling ofling dysmorphic . . W

) ling withling MCM2 (Figure . comparedto adjacen in allcases noted

C and

ith the a trong cytoplasmic labelling albindin,calre ,

more prominentlabelling of neuropathologydiagnosis of

DCX thedysmorphic neurons of the intense nti mainly spared tau from a populationof and intenselabelling 4 (Figure4h ionof consistent - g , ; occasional ; pS6 (ser235/236 andser240/244)

)

and AT8phosphorylated for

.I and nes

inthe MVN markers

(Figure4b studied

nterneuronsof

in smallcells and inflammatory interneurons labellingof scatte

(Figure4 tinin,parvalbumin andNPY

neuronswas notedin one )

tin in some cases labellingwas observe but demarcati , , mutant apart , tcortical

) dot T, only bipolarcells with long

. labelling accumulation

CD34 was noted

m althoughthere was -

likepositivity in the from onefrom case ) ) in contrastto Ki67 occasionalVC in adjacent cortex normal

IDH1 for s -

for pS6 for and w ( theng nodular positive C red pyramidalred

p62 (Figure 4j with small ase 2)ase , ATRX , AD

(Figure

OTX (Figure 4l hite , KCC

the the -

cells tau was tau 1 to to d with and 2 2g ,

, ) . )

classification was ( associatedwith epilepsy 2014 In Discussion long controlsas well as published dataon immunohistochemistry MVNTand occur not in all FCD or glioneuronal ty other glioneuronaltumours and 8 glioneuronal tumours (Table 3 surgicalMVN in any Case theOf 33 genesstudied MOLECULARGENETICS SOX2(Supplementa labelling no but labelling with KCC1 (Supplementa occasionallabelling of oligodendrog SOX2( not but strong MVNTto (Sup reported calbindinpreviously as noted ganglioglioma SOX2(S 24 pe II samplesIIpe ) . S - added to added to term epilepsyterm in 8 MVNT ubsequently Tbr1

Supplementa OTX1 (SupplementalFigure 1h , (Table 3)

Huse

upplementaryFigure 1c). There was little evidenceof and ( primarilyin the p 43

90%of FCD none but of the glioneuronaltumours. label )

. . l Supplementary T tested T ( ia (

s the the 33 D Recurring plementary1f). Figure In etal.,

forOLIG2 was seen(SupplementaryFigure and ysmorphiccells in ganglioglioma ) Supplementa . l ing of intersting . listof

AcceptedNomutations in Article

, We present the pathologyWe in

five the

ryFigure 1m,n,o). ry described tencases of Supplementary ,

white with less frequent findings Figure 1i)

patterns of majorityof , more

. ;

Identical SUFUmutation wa matter

Table 2 coined the term (SupplementaryFigure 1e it ry caseshave been reported ialwhite matterneurons was noted(Supplemental in mutation

Figure 1j).pS6 expressionin

or MVN MTOR glioneuronaltumours CNSneoplasms

neurons tumours Mapb1b; ).

DEPDC5 Table 3 related SNPsin W ) T

temporallobe wh ,

s an e compared the are summarised and synonymous ,DEPDC5, FGFR1, MYB ry

drare positive neurone

Figure 1l).

anovel . SUFU

tu and small glialcells

OLIG2 was restricted to mature Of note,Of a ‘multinodularand vacuolati

D moursand malformati and s presents in oneFCD type butcaseII not in a furthe EPDC5 infrequentl

was identified SMO low inthe SNPs In In fetal , -

) r r ten , NPRL3 grade grade , while , other the common

was confirmed with Tbr2,OTX1 and (

ite matterwith mild MCDtype II ( E3SNP MVN SMO 62

7 recently ,

) 1d)but strong labellingwith 17

occurring cases,including a case and

labelledwith MCM2contrast in

TLEwhite mattershowed developmentalcontrols T to FCD T typeII samples)(22 comparedto finding

neuronal

, The and nuclearlabelling of

39 varia

( also in also Supplementa s , revised2016 W

or TP53 genetic

68 Case wereobserved with ons ntwas infound 5/8

BRAF ) with p62 occurredin allFCD insome MVN ,

lesion an ng neuronal tumour’ asso werepresent in all 2 d this new d this entity , and

wereidentified andin EZH2 - appearing , , mainly ciatedwith

ry aspreviously

Figure 1g) Figure 1k) s ins SNPs HO T only T

strong

, did

II reported hamartomaassociated with gela variationsin noted frequentlyin MNVT andFCDbut not other glioneuronal t SNPs ( abnormalities ofLEATs SOX2, TBR1, OTX1,KCC1 and GFAP∂, reportedcases undisturbed laminar cortex activity following study. includes This corticaldevelopment Aspreviously noted significanc withcases conventionalganglioglioma composition. characteristics associated tumours(LEATs) classification MVNT NEOPLASIAMALFORMATIONOR share lesionsencountered in epilepsyseries( unrecognised neoplasm remainsIt possible that mechanismsof epileptogenesis investigations into presentingwith findings previously 15 B

, include 25

histological in SMO , hasbeen ,

(i 42 confirm

in FCDII i incompleteresection s i) e. reportedin 2007as an unusual , . the novelmutations in SUFUand EZH2

lack of 44 A

SMO

We incl We ofCNS tumours , lthough

and DEPDC5 , being strikinghypo , (

late 47 39 (v) retainedexpression of immature, developmentally regulat

their grouped

( Accepted Articlesimilarities. , , areceptor in theShhpathway have beenrecently shown hypothalamic in the 5 expansiveinfiltrative or growth pattern

68 50

, onset ( (i) ude the

misd 24 56 or d indolent these lesionsare ir ) ) , frequentCD34 ,

the lack

ysplasia than ia as apattern of of Further ,

) gnosed gnosed myelinationof (iv) in ,

(

in common ep firstreport of a case with mixed

57 some .

behaviour , , (ii)no

support F align ilepsy

ofany comparable ) eatures through

NGS include t

ic seizures h

features ofMVN

istological f asde more

, overtincrease in cellularity growth on typicallyin adulthood , inthe present study ofa -

a rare,it isprobable they positivity a including

s malformative nature ,

incommon with otherlow the application the o with corticaldysplasia (FCDIIB)

s listeds in gangliocytomain the apredilection for the neoplasm,supported by evidence in thiscurrent (v and

myelinatingconditio with developmentalanomalies involved white vacuola potential i ) absence ) of any

localisations in ( predominant 22 eaturesin comm , although , , )

serial BRAF and a

a ted ted Supplementary nd T commonorigin

align diffuse DEPDC5 MRI

mixedneuronal and glial V600E

matter s

these mutations ofare immunohis

showi

with nodules ‘sitting’in invol . . hav We morewith a deepcortex/subcortical region or

featuresof both ofthe known gangliocytoma n ,

revised reportedregrowth, even

, temporallobe umours s cell have been previously

ve on betweenon MVN

MYB

which maytorelate

and ng ( or 7 - ment of ment the temporallobe grade ) Table 3 ,

lineage

conspicuous recurring orother morecommon and we identified e extended NPRL3 tochemistryand NGS and

2016 WHO .

Copynumber malformationof

than epilepsy and

FGFR1 genet ) with ) which they and

( mutations ed proteinsed as

MVN synonymous

45 NPRL3 M true

potential

RI ic )

uncertain . . Our T and T mitotic

- mutation an T and T

SNPs two

FCD

in s .

GLIAL DIFFERENTIATION remnants support for their derivation radialfrom migrating progenitorcells incortex deeper compared to outer cortical layers. deepercortical layers. This VCderare observedin ganglio ( expressed inlayer Vpyramidal neurones precursors OTX1 is stronglyexpressed tem cortical layers gestation duringdevelopment e I ( SOX2and CD34, whi 68 also nodulesalpha for inareas adult the brain orregions undergoing structuralplasticity MAP1b consis expressed in typicallyweakor negative. labelling for 17 including Previousreportshave emphasised the LINEAGE progenitorcell type considered VC of deficiencies mmunohistochemistry 20 35 xtentof the MVNT ) , , , 28 poralcortical in 29 41 beenpreviously shown , is linewith ourobservations. tent , ) , which , ishighlydevelopmentally regulated 39 . . Theconsistent e 57 reminiscentof

HuC/HuD , ( lyexpressed in ourseries. W ANDMATURITY

) 16 ivedincomplete from maturationor migration of progen ( 68 . not to

some in oligodendrogliallineages

) migratingneuronal precursors w ) . .

) . Ourstudy supported Vand VI

aswe also observ thinning - reflect Accepted internexin,intermediatean neurofilamentexpressed in Article anti

, . glioma orthe (

compared to 60 In In murine cortex ch has beench has -

neurofilame , for for OTX1 balloon cellsin

12 ) Expressionof

. couldone be

particula in MVNin in the forebrain and ANDMYELINATION OF VACUOLATINGOF CELLS

)

primarilyin earlymigrating andspontaneous seizures

We also notedWe showed dysmorphicneurones ofFCD mature

ed consistently ntantibodies cellproliferation T rly

. , evidence for

Inmature human cortex ( homeobo 39

enoted however preferential in thetemporal lobe FCD (

, 46 explanation predominantfor the location DCX neuronalmarkers strikinglabelling of VC,highlighting the ( 29 68 ,

( , 51 ) 61 , as ,a microtubule 38

re butmore variable expressionof nestin , x gene )

( intense proli 55 ) .MCM2 labellingof previously particularly portedin CNStumours,

matureneuronal differentiation as wellas immature cellsin FCDand I II

butnormally retained in neurogenic Focal expressionimmat of , ,

70 but ferative layersofthe neocorti

) OTX1 appearedmore

.Also the

expression ofstemcell m cellcycle morerobust expression with

TBR1 expressionin VCis further neurones ( 3

) reported as ( . SMI32)although NeuN was 3

- In developingIn the human brain isexpressed in midto late , associatedprotein )

. NeuN OTX1has beenshown be to (

OTX1 orpossibly subplate 20 (

distinctive i 6 arrest tors )

destined for 6

and

in a ballo ) inthe lesional cells .

nullmice show

Strongexpression of destinedthe for pronouncedthan ure markers,

could suggest inpathological a MVN LEATSand FCD neuroblastshas on

MCM2 positivity cells T

deeper (

39 (MAP) subcortical

ofVC with arkers is cal of )

but MVNT (

39 t

hat

not ( , 7 ,

the the temporallobe with 18 ofthe 24 reported casesaris confirming MVNT of documented years44.6 Fromthe periods g oper C contours, and FLAIRand satellite nodules Characteristicapp NEUROIMAGING matter associated a novel vessels observation myelination types furtherhypothesis contextof oncogenicrole in glioma growth 17 both the PAX6 andnestin also arguesa for mi preferential tumours including associatedwith we parenchyma GFAP rowth ombinationsof these , aro observed 24 ativelyother from LEATs

und 30years (OPC)

in MVN with myelin loss

over18 months , 39 subty in this

( reminiscentof VC

11 24 cases24 ofMVNT so , the overall .Although

l ( with increasedOLIG2 cellsin deep the cortical layers and ) metimeswith corrugated a outer margin in MVNT , abellingof aswell 9

( in pe of of pe

distinctpopulation but in keeping with previousobservations 23 ) subventricularzone ) T highlighted T series

andconfirmed in our series

just and preferentialinvolvement of the grey MVN

)

awayafrom main lesion

with

with reduced

, maybe encounter

Acceptede Article is that

. mild malformation CLINICAL PRESENTATION

arances reportedin lessthan of half reported cases even as the smallercells in MVN However .

epilepsywas in present in thisand previous reports a l

developmentally .

imaging smaller also drawscomparison with MVNT developmentalmigration patterns (

ess GFAPδ 7

myelinationan 24

a variable reactive glialcomponent

consistentexpression reported , originate OLIG2from , gliomasand FCD )

we include

as intralesionalcells s features mayen

( is

64 of

also supported by ( normallyrestricted to 65

) intra ed ined the paediatric age ,

xedcellular composition in MVN ofcortical developmentin frontal ) however (

7 aswell as some

arrestedor partial MRIof MVNT , - nal

lesional d

17 the youngest the (Nagaishi,Yokoo et 2015),al.

. T labellingof VC for

, differentiation

24 his transcription hasfactor a recognised allour surgical samples it in MVNT T , able the . ,

cells IntervalMRI alsohas shownof lack a for for OLIG2 beenhas previously reported ( ratherthan the VC - 24 in positive -

) withaverage an age of onset of seizures ( white matter junction fol ,

longer 7 (Fukushima,Yoshida et ) reactive themean age atthe

, include has beenhas clustering VC of along as evidencea for inghere, with mesial a component

24 case malfor

OLIG2 glia future

of OPC ) maturation (

.However

19 ( o pre - 52

,operated residing group. ligodendroglialproge

) myelin basicprotein gliosis multi mations .Indeed a )

distinctionof MVNT . merging operative expression

( 63 -

nodularity MVNT

, , ( )

27 forp . Recent . description resulting

using

superficial ( lobe

at sixyears

29 stemcell niches

hasbeen

) an n occ glial hyperintensityT2on . Uniform observational

, Furthermore,

S6, PDGFRβ,S6, timeof surgery is

predominate in

36 isnoted in adjacent

epilepsy lowinggyral isoformGFAPδ -

cell al. 2015).al. ) asionally (

7 in and ,

68 component white defective labellin . nitorcell

One pre astrocytic ) , , gliotic

the the - g of g

( of 7

indolentnature of MNVT Sincesubmission of this study, furthercases and serieshave been reported,attesting to the Noteadded inProof indicatean a mutationsidentified lobeolder in conclusion,In MVNT operatein MVNT astroglia with cell evidencesome for investigated explorWe activitydrive seeding include AD,sparing ofVC and immunohistochemistry. therewas striking a absence of tangles or phosphorylated with tau AT8 increasedor effectanbe of ganglio formation andp62 VCseveral for rea or could An EPILEPTOGENSISAND NEURODEGENERATION asso cas formation andsubiculum have been eleve mixedcellular composition and expression of relative o es which could potentiallyenhance hippocampal lder ci n

be explainedbe by ated hippocampalated sclerosisis

( glioma

a functional (

immaturity ageat ( , 31 48 ed functional inactivity. functional W 17

origin froma )

n )

in relatedpathologies patients , neuronal , otherpotential . or focalinflammation 24

(

mTOR

manifestation of indirectly ,

but , 39

Accepted Articlesons :

accumulation

cation 43

(

in LEATSor 37 , disconnection

areuncommon warrant further with seizure ,

68 over their excita 54 , nodules taufrom accumulation Indeed n 49

)

) accumulation ofhyperphosphorylated tau/neurofibrillary tangle 1 chloride cotransporter it couldit aberrant ,indicating - )

, activity , pred

pro bility 4 reduction , , 10

- even ominantsubcortical epileptogeniccellular alterations

epilepsy occursearly in morphologicallysimilar malformations.Ourstudies support eexplored acquisition of neurodegenerative changesin the ( , s. ( refl 14 - 67 40 like’extensions of the tumour along the hippocampal ( ( progenitor celltype of 2 43 The

) previously variable. , ) in ectred

) butdistinct lesions in the .By contrastin MVNT an as pro . s 53 )

- , , thatwe demonstratedin MNVTwith pS6labelling,

depth yappear ofinhibitory interneurons , enhance

in 58

oldest MVNT uced excitabilityo

- , from the adjacent cortex epileptogeniccellular alterations

investigation 59 described,

a range of

dysmorphicneurone ) (KCC1/KCC2)

indolent d ( Supplementary post

compared otherto vul

locationwith

epileptogen MVNT nerability degeneration.to This

- was noted.Possible explanations duringdevelopment mortem

mainlypresenting the in temporal and lackany . ( immaturemarkers which could

68 ,although p62 was prominent, ) f

VCif tau phosphorylation is andwas in striking five ofour im casewit i

city,although presence of in MVNT limited within lesions balance table3) s ofs FCDand epileptogenic preventingtau

of of the h high Braakh high stage cortical s . . We

as previously

. in

, common

VCin that

, identified

immature connec could keeping lesions

could tivity

consistentwith those guidelines. Journal’sposition on issu Avanita Prabowoskilful for technical assistance. confirm We thatwe read have the Foundation, and BankTissue at UCL is funded Health’sNIHR Biomedical Research Centres scheme. funding TheEpilepsy Society Brain wasat undertaken UCLH/UCL whoreceived a proportion of funding thefrom Department of work This is supported by Medic the Acknowledgments

Accepted ArticleKIKA(Stichting Kinderen Kankervrij; andAB) Stichting Knip. areWe gratefulto

by Epilepsy the Society. EA is supported byAMC the alResearch Council(grantMR/JO127OX/1). work This cor is shown synaptophysin (D) MAP2, were typically (VC) ( Figure 3. M forJ, L, microns bodies. granular of appearances typical more showed lobe temporal like cells cresyl violet on between (arrowhead) border ( (arrowhead) were seen of vacuolation prominent white (SMI94) staining basic protein Myelin var cortex adjacent to compared nodules cortex adjacent w layers cortical deep the on encroaching nodules the highlights of th medialpart mm predominan although layer, in subiculum the in cells of islands nodular 1. (D)Abnormal white matter the in (arrowhead) like patterns nodular and (arrow) myelination poor of regions diffuse confirms protein basic SMI94/myelin (C) matter(arrowhead). white the into extending junction white matter Case cores and gyral the features (A) Macroscopic Tumours ( Figure 2. lobe temporal sequ on growth the lesion in on hyperintensity noted par 1 (A) Case Figure 1 Figure Legends A) NeuN staining showed labelling of normal interstitial neurons in the white matter white the in neurons interstitial ofnormal labelling showed staining A) NeuN L )

iable patterns with reduced MAP2 labelling in some nodules (arrowhead) compared to others (arrows). others to compared (arrowhead) nodules some in labelling MAP2 reduced with patterns iable tical nodules. (F) TBR1 (F) nodules. tical ahippocampal gyrus and and gyrus ahippocampal In some cases alignment of the atypical cells along vessels was noted in the nodules the in noted was vessels cells along atypical the of cases alignment someIn secti ences, highlighting a highlighting ences, 1 confirms lack of myelin in in myelin of lack confirms 1 matter regions are regions matter over8 year

in the nodule the in ons of the temporal temporal the of ons Macroscopic Neuro MVNT euroimaging

( MRI, as MRI, as reported in the original cas original the in reported as right posterior temporal posterior right .

nal nal .

s ) (G) Neuropeptide Y in Yin Neuropeptide (G)

e specimen w e specimen tain gives gives tain

Bar = 1cm for =Bar 1cm s. (B) Case 8. Abnormality was observed Abnormality 8. Case s.(B) coronal coronal , N

shown in (D) (D) in shown And Glial Marker Expression In Vacuolated Cells( Vacuolated In Expression Marker Glial And weak or weak or focal

and =50 microns for K (approximate based on original magnif original on based forK (approximate =50 microns and

Accepted Article s, a

Features Of Multinodular And Vacuolating Neuronal Tumour. Neuronal Vacuolating And Multinodular Of Features

nd Low Power Histological Features Of Features Histological Power Low nd

and cellsand compared to the adjacent white matter. adjacent the to compared

translucency of the the of translucency

populated by single by populated of the temporal lobe specimen from Case 1 show areas of areas show 1 Case from specimen lobe temporal the of hyperintense abnormality and the cortical, white matter interface in the left the in matter interface white the cortical, and abnormality hyperintense neu the FLAIR sequence. (C) Cas (C) FLAIR sequence. the impression the more often often more tly in the subcortical region subcortical the in tly

hip

macroscopic images in A and E; = 3mm for B, = 3mm E; for and A in images macroscopic pocampus on on pocampus

at with more eosinophilic cytoplasm eosinophilic more with some . (E) Overall, Overall, (E) . n overlying n 21 months 21 months

Case lobe in in (arrowheads Case Case

gyral cores (arrow) a (arrow) cores gyral negative in MVNT in Case or vacuoles surrounding the cells. the (K) surrounding vacuoles or e report

as a diffuse cortical abnormality, abnormality, cortical diffuse as a of overall reduced cellularity, particularly cellularity, reduced of overall 2 show 4 at low power shows reduction of labelling within the cortical cortical the within labelling of shows reduction low power 4 at white matter white 4 scattered

coronal coronal normal following following highlights highlights

reduced labelling with synaptophysin synaptophysin with labelling reduced

(top half of figure) and half (top (arrowheads) ( 45 e 5. Initial FLAIR MRI FLAIR Initial e 5. s ). shows shows

islands of grey tissue in the white matter of the inferiorof the matter white the in tissue of grey islands

a ganglioglioma a ) (H) MAP2 staining in in staining (H) MAP2 ) -

FLAIR showing appearing appearing

cells with a neuronal/ganglion neuronal/ganglion a with cells (arrowhead) the the abutting

. nuclear nuclear in the right mesial temporal region with with region mesial right temporal the in

(B) Luxol fast blue/cresyl violet stained section from section stained violet fastblue/cresyl Luxol (B) Case nd pale pale nd but with minimal mass effect; mass minimalwith but initial scan initial . Multinodular Multinodular

marked signal ch signalmarked Labelling of VC with (B) neurofilament, (C) neurofilament, (B) with ofVC Labelling ith reduced labelling compared to the the to compared labelling reduced ith 1,

( cortex. labelling of t VC and strong strong and VC of t labelling N ,

present on both sides of the pyramidal cell sides pyramidal the of both on present myelin were observed in MVNT MVNT in were observed ). In ). In hypomyelinated hypomyelinated , VC .

with dysplastic neurones and eosinophilic eosinophilic and neurones dysplastic with (E) Macroscopic appearance of fixed fixed of appearance Macroscopic (E)

)

sequences at sequences . the white matter (lower half of figure) figure) of half matter (lower white the

(E) Case 6 shown with T2 and with shown 6 Case (E) C (F) Synaptophysin labelling in in labelling Synaptophysin (F) ase 8 Case Case - poor nodules. ( nodules. poor

which di which And And C, D, F, G, H ; =0.5mm forI ;=0.5mm G, H F, D, C,

, Occasional 8 in the temporal cortex shows shows cortex temporal the 8 in

a focal area in the the in area focal a ange in the left temporal lobe, lefttemporal the in ange Vacuolating Vacuolating ications).

breakdown/cavitation in in breakdown/cavitation

for nodules at the cortical cortical the nodules at

d not show significant notsignificant show d presentation highlighted a a highlighted presentation ,

was observed but the the but cell appearance appearance cell

smalloligodendroglial ( Case Case

binucleated cell binucleated ) little change little change

The abnormal abnormal The

after H&E staining after H&E Neuronal Neuronal cytoplasmic 4) vacuolated cells cells vacuolated .

(M). The The (M). mesial in the the in

(F) in in and and C was was

ase 4 ase ; = 100

FLAIR s ase ase 5

(I) (I) - .

matter neurons in Mild MCD. Mild in matter neurons cells smallglial and neurons matter white single the of labelling occasional cells seen. oligodendroglial small ofthe labelling only and neurons white matter single glialcells. ofsmall labelling weakcytoplasmic and in neurons a in positivity variable nuclear cells (inset). neuronal intense tissue and adjacent with border sharp a with parenchyma and tumour feature or gangliogliomas cells of neuronal staining) cases. of proportion shown). cases (not further (A) Tbr1 Development ( Neurones Matter Increased White With Epilepsy : Comparative 1 Figure Supplemental m original positive. MCM2 nuclear left (insert overlying cortex dot from occasional apart (L) antibodies. MVNT pathology in tau forphosphorylated labelling (I)AT8 VC. of the labelling cytoplasmic ( transporter chloride (H) Cationic parvalbumin. for ofVC labelling showing strong mor normal of lesion the in interneurons (c MVNT one of nodules right). cell (inset bipolar occasional and lesion the in noted cells were mainly VCwere (E) pos doublecortin (D) Occasional VC. regions with the in evident processes were and cases between staining the in variation showed CD34 VC and nodules ofthe labelling strong showed (A) Figure 4 magnifications). J G, H, nodules SMI94 (K)PDGF whereas VC OLIG2 with show ; low power OTX (G) with noted labelling PAX6 was negative in vacu in was negative PAX6 s ) = 40 microns (I) and 140 microns (B, K) and 200 microns (E and I) (approximate, based on original original on based (approximate, I) and (E microns 200 K) (B,and 140 microns (I) and microns ) =40

for tau accumulation ( accumulation fortau lesion the around gliosis ofthe labelling more extensive with pattern opposite for myelin basic protein, basic protein, myelin for ; (inset) PDGFRβ did not label the atypical ganglion cells ganglion atypical the label not did PDGFRβ ; (inset)

. Immature Immature . (s

i of dysplastic ganglion cells which was variable between cases variable between was which cells ganglion of dysplastic n agnifications). , ee also

gangliogliomas showed strong nuclear labelling in two cases as shown, but negative labelling in two in negative labelling but shown, as cases two in labelling nuclear strong showed gangliogliomas m and dense cortical and dense cortical inset inset ild

Type VC

MCD.

Figure 2) were not positive with both pS6 markers pS6 Ser240/244 (shown here) and pS6 Ser235/236 pS6 and here) (shown Ser240/244 pS6 markers pS6 both with positive were not

(top) (top) nestin negative but occasional positive cells were seen (inset left). Small nestin expressing nestin expressing Small left). (inset were seen cells positive occasional but nestin negative II And Interneuronal M Interneuronal And (C) Accepted Development ) And Article (F) (H)

ase 4) ase multipolar GFAP multipolar SOX2 in gangliogliomas showed granular cytoplasmic (or peripheral peripheral (or cytoplasmic granular showed gangliogliomas in SOX2 Bar = Bar MCM2 labelling in gangliogliomas highlighted inflammatory cell component with with component cell inflammatory highlighted gangliogliomas in labelling MCM2

There was no labelling of OTX1 in the white matter neurons with OTX1 in OTX1 in with neurons matter white the in of OTX1 labelling wasThere no - , also , (B ) showed strong strong ) showed cytoplasm the in positivity ike and (H) MAP1b (H) and for SOX2 was note SOX2 was for ) OTX1 in ganglioglioma showed cytoplasmic positivity of dysmorphic neurons in neurons dysmorphic of positivity cytoplasmic showed OTX1 ganglioglioma in ) arrows). ( arrows).

small 50 microns (A,D,E,G,H,K,L,M) = 20 microns (B,I,F,J) (approximate based on on based (approximate (B,I,F,J) =microns 20 (A,D,E,G,H,K,L,M) 50 microns arrows cells ofsmall labelled nuclei but (VC) cells lated as well as demonstrating as well as demonstrating showed membranous labelling of the VC of the labelling membranous showed R

TBR2 in fetal cortex showed labelling of immature cells in the germinal the in cells ofimmature labelling showed cortex fetal in TBR2 beta highlighted small multipolar cells in the nodules but not the VC the not but cells nodules the in multipolar small highlighted beta

showed cytoplasmic granular s granular cytoplasmic showed proportion of the ganglion cells ganglion the of proportion Labelling In Ganglioglioma Cases, Ganglioglioma In Labelling J) VC J) showing the ed the showing al Controls Of 13 Gestational Weeks. Gestational 13 Of Controls al δ phology but the VC the but phology arkers In arkers In

cells in proximity to VC), inset (bottom) GFAP conventional isoform conventional GFAP inset VC),(bottom) to proximity cells in

scattered scattered were strongly positive with p62 and (K)anti and p62 with positive strongly were . (I) GFAP .

d. (J) d. highlighting the regions of involvement at l at regions involvement of the highlighting

Equivalent To Equivalent Multinodular Multinodular (I)

In In I , δ n neuronal neuronal

ge of the nodule the ge of but as shown here in in here as shown but

m highlighted small glial cellsglial primari small highlighted m

ild ild the ild ild of uncertain significanc uncertain of

(K) MCD, the neuronal cells were not OLIG2 positive OLIG2 positive not cells were neuronal MCD, the MCD, occasional weak cytoplasmic labelling of the ofthe labelling weak cytoplasmic MCD, occasional were negative (arrowed were negative

diminished myelination in the white matter matter white the in myelination diminished phosphor . positivity

And Vacuolating Neuronal Tumour Neuronal Vacuolating And Mild MCD Mild (E) (E)

a marked sparing of the VC and nodules VC and ofthe sparing marked a taining Calbindin showed strong labelling of the ofthe labelling strong showed Calbindin . (G) TBR (G) . ( F . (D) . ; - ) NPY staining labelled labelled NPY staining S6 labelling in mild MCD showed MCD showed mild in S6 labelling TLE TLE . NPY

(arrow) but nuclear labelling was not a not labellingnuclear was but (M) A high proportion of VCwe of proportion high (M) A / Case

and (L) and Malformation Of Cortical Of Cortical Malformation OLI Case Case /Temporal Lobe In Temporal Lobe Temporal Lobe In Lobe /Temporal 1 showed labelling ofwhi labelling showed 1

showed

G2 was negative in the atypical atypical the was in negative G2 labelling of dysmorphic dysmorphic of labelling within the lesion the within 4 with Alzheimer 4 with e . 5 Bar = Bar (N (insert left) (insert , K

itive (DCX) (DCX) itive prominent mu prominent CC1 did not label the white white the label not did CC1 /

s. KCC1) with disti with KCC1) in insetin reduced reduced

80 microns (A, C, D, F, D, C, microns (A, 80 - (J) mitochondrial mitochondrial ly in the nodules at at nodules the in ly

ow magnification ow magnification Nuclear labelling of labelling Nuclear ). scattered scattered ; in cont ; in cytoplasmic cytoplasmic

(G) Case 3 Case (G) VC were seen were VC labelling in in labelling . (B) SOX2 (B) . ’ s disease s disease ltip

m nct nct olar cells olar ild rast the the rast te matter matter te .

(L) MCD MCD

re re on on . (C)

a ,

magnifications). G(appro on 200microns and E in microns =100 (A,B,C,D,F,H,I,J,K,L,M,N,O); SOX2 showed zone periventricular the in matrix and strong strong a strong labelling

, pr , ed

and developing white matter. matter. white developing and in the germinal germinal the in cytoplasmic cytoplasmic matrix

was labelling o labelling seen. Bar = Bar seen. (N)

OTX1 in developmental controls controls developmental in OTX1 f the germinal matrix cells. (O matrix cells. f germinal the ximate based on original original on ximate based

50 microns

) With With GROUP

Antibody

Accepted Article (s ource )

(mins, temp Antibody Pre - treatment di erature) lution ,

PATHWAY MTOR VE* DEGENERA NEURO NEURONAL* INTER MENTAL* /DEVELOP STEM CELL MYELIN GLIAL/ DENDRO OLIGO ASTROGLIAL MAR NEURONAL LAYER CORTICAL MARKERS NEURONAL MATURE

KERS* - -

CD34 SOX2 (Abcam) PDGFR beta PAX6 Reelin SMI94 OLIG2 delta GFAP GFAP MAP1B N200 OTX1 TBR2 TBR1 MAP2 NFc ( Synato SMI31 SMI32 NeuN ( pS6 Ser240/244 ( Mitochondria p62 APP AT8 KCC1 Bioclear) KCC2 (Sigma Neuropeptide Y P Calretinin Calbindin Signaling Tech. Doublecortin Nestin DAKO Cell Signaling Tech. Abcam arvalbumin

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Synapticproteinsynaptic vesicles / Neurofilament kDa protein)/axons (phosphorylated 200 axonssome pyramidal andcell bodies Neurofilament (non Neuronalnuclear nuclei and /neuronal antigen cyto phosphorylated Anti amyloid precursor protein Phosphorylated microtubule tau associated protein Na+/K+/Cl K+/Cl Neuropeptide neurones /GABAergic Calcium binding protein Calcium binding protein Calcium binding protein associated protein Developmentally microtubule regulated neuronal expressed Intermediate filament;developmentally regulated Stem cell marker sex oligodendroglial precursortypes cell Platelet receptor derived growth factor paired cells in development cortical E Myelinbasic protein Oligodendroglia factor transcription lineage astrocyt GFAP developmentalisoform, regulation GFAP fetal cortex. expressed development in Microtubuleprotein associated 1 Neurofilament 200/ subset projection layerneurones. of V/VI Orthodendicle 1/ homolog cells T derived intermediate from progenitor cells (IPC) T processes Microtubuleprotein associated Neurofilament cocktail Sequestosome xtracellularprotein matrix - - box brain protein/ box brain protein/ - determining region Y -

mitochondrial antibody - / astrocytes

- Cotransporter/GABAergicneurones box protein; es, stem cells

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Cotransporter isoform

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/ neuroblasts targets specific cargoes for autophagy cargoestargets for specific -

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projection neuronesprojection phosphorylated 200kDa protein)/ nuclei marker nuclei marker

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- interneuronal markerinterneuronal markerinterneuronal i box 2 nterneuronal markernterneuronal

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expressed in Retzius Cajal / progenitor/ cells

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MARKERS MUTATION TUMOUR AND CELL CYCLE ACTIVATION Transduction Oxford Lab., Heidelberg, Germa Sternberger, US Maryland, pH6 buffer Citrate SC Sodium H UK); Peterborough, (VectorLab, Keynes, UK); Keyn (Leica, Milton diluent and enzyme concentrate these cases in achieved reported cellsglial aspreviously examined cases remaining ;for sections of availability limited to due marker each with were studied cases 8 to 4 *between with indicated antibodies of groups the in markers For (MVNT). tumour of immunohistochemi List Table 1.

ATRX IDH1 Bioscience BRAF V600E Ki67 MCM2 ( pS6 Ser 235/236 in all cases all in Cell Signaling Tech. ER2

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- in this series was used to label pericytes as well as small multipolar NG2 multipolar small as aswell pericytes label to used was series this in cal V600E mutation Serine/threonineV600E to Nuclearprotein/proliferating cells replication Miniprotein maintenance chromosome licensed for /cells phosphorylated (is SNF2 helicaseATPases) family of /gliomas and Alpha syndromex retardation thalassemia/mental gra Isocitrate Dehydrogenase grade 1/low high secondary and Upregulatedbenign human in tumours malignant and 3300 Vector’s citrate Vector’s 3300 , 55

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(Leica, Milton Keynes, UK); H UK); Keynes, Milton (Leica, , California,US,

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; Autogen Bioclear Ltd,Wiltshire, UK

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ER2, 1:500 ER2, 1:64 ER2, 1:50 ER2, 1:200 ER2, 4ºC) H 4ºC) H all based buffer pH9.0 pH9.0 buffer based , Watford, UK Watford, , - - 3301, 3301, 1

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Epilepsy Epilepsy Seizures Seizures Speech arrest Seizures/ seizures symptoms/ Visual difficulties Breathing Seizures epilepsy poral lobe Seizures/tem Seizures Presenting Presenting symptoms

partial seizures, GS=generalised seizures Alzheimer’s’ disease,AED= anti following surg Table

2 21 47 65 NA 10 years’ many ‘epilepsy seizure 39 (years) onset : Clinical: details of casesin current series, location, salientMRI features and outcome Age Age

of s

e 41/F 54/M 6/M 48/F 67 /F 62 /M 27 /F 32 /F 59 /F 55/ ry /Gender surgery surgery Age at Age . M (PM)

Case 1 was previously reported (Rat

Accepted Article

epilepticdrugs,

temporal pole Temporal lobe and lesion Right mesial temporal hippocampus fusiform gyrus and parahippocampal gyrus, Right temporal lobe, Left temporal gyrus Left middle temporal temporal lobe Right posterior parietal Right occipital lobe hippocampus Left amygdala and amygdala hippocampus and parahippocampal gyrus, Right temporal lobe, amygdala hippocampus and parahippocampal gyrus, Left te Structures involved Localisation

mporallobe,

lesion

NA=notapplicable, RTA = road trafficaccident, PS =

ilal al., et 2007). Post mortem : whole brain resection of amygdala hippocampectomy and Left temporallobectomy, resection of amygdala hippocampectomy and Right temporal lobectomy, resection of amygdala hippocampectomy and Left temporallobectomy, Temporal lobe resection Temporal lobe resection hippocampectomy Tem hippocampectomy Left temporal neocortex, craniotomy Lesionectomy /awake Lesionectomy poral lobe, Type of surgicalType resection

PM post = mortem, AD =

Braak stage V AD lobectomy. temporal following Psychosis Migrain verbal learning. Impaired non Depression operatively. memory post Declining verbal weakness operative facial psychosis/post Post None None mesothelioma. Cause of death diagnosed at PM neurology Any other - relevant ictal

NA years][14 recurrence on MRI Seizure years][6 recurrence Seizure years][9 tumo No recurrence of seizure per week)/ (1 Continued seizures data Limited follow up Recent surgery , MRI no progression on Partial resection but [10 months] progression Seizure [17 months] progression MRI but no residual tumour on Seizure on MRI but no progression /Residual tumour Seizure years][2 stable seizurefree /lesion deficit and No neurological following surgery. following surgery. [ [Period of follow 2 years] control/tumour - 3 nocturnal

recurrence recurrence

Outcome ur Seizure

- - - - -

free/No free/ free free /No free /no up]

Gene

Exon

Position Accepted Article (hg19)/(hg38)

AA change

Nucleic Name

SNP database no

Mutation effect

Total cases MNVT

[case number]

Positive FCDII cases

Positive mixed lowgrade glioneuronal tumors NPRL3 PIK3C DEPD SUFU EZH2 TP53 SMO CIC C5 A

20 11 4 6 3 6 9 6 8

[chr10: [chr10: [chr7:g.148525904 [chr19: [chr19: [chr22: [chr3: [chr7:129206487] [chr7:128846328] [chr7:129205177] [chr7:128845018] [chr3:178922274] [chr22:32179850] [chr17:7579472] [chr16:142825] [chr16:92827] 108 (1018) 119 (215) 24 (1164) 74 (4533) 69 (553) (hg19)]

179204486]

- 102599540] 104359297]

- - - 42294897] 42799049] 31783864] 102 26 17 43

Accepted Article

A I P    Intron Intron Intron Intron D G (185) (388) I (1511) S (340) R (72) R - - > H > G

c.538 c.1164G>C c.1018G>T c.453 c.553G>C c.215C>G 102A>G c.1032 c.1060 4 17C>A c.484 3G>C - 26C>T 3C>T - - -

rs25416 rs13828 rs23024 rs34135 rs10425 rs22286 rs27030 rs26998 rs10520 762470 762469 766190 004294 756833 COSM3 COSM3 COSM3 COSM4 COSM3 067 18 27 22 17 91 96 23 6

mutation of mutation of significance si unknown unknown Missense Missense Polymor gnificance Intron Intron Intron Intron phism Silent Silent

1,2,3,5,6,8,9 1,2,3,5,6,8,9 1,2,3,5,6,8,9 [2,3,5,6,8,9, [1,3,,6,8,9,1 [2,3, [all cases [all cases [all cases [all cases tested ; tested ; tested ; tested ; ,10] ,10] ,10] 10] [8] [2] 0] 5 8 1 1 8 8 8 7 6 5,

6,9]

20/22 22/22 20/22 21/22 22/22 19/22 14/22 0/22 1/22

7/8 7/8 8/8 6/8 4/8 0/8 6/8 0/8 0/8

generation sequencing Table3. Mutations and recurring, common single nucleotidepolymorphisms [chr17: Accepted Article 7676154]

identifiednext by 1,2,3,5,6,8,9 ,10]

Neuroscienceand biobehavioral reviews.63:177 (2016)Inflammatory mediators in human epilep 14. Naturereviews Neurology.12(7):379 13. somatosensory cortexin Otx1( Acampora SimeoneD, A, Spreafico Frassoni R, C (2002) Morphological organization of 12. deltaexpression inhuman astrocytic tumor. Neuroscienceletters.463(3):182 11. pathology.41(7):1005 Vacuolating Neuronal Tumor:A Rare Seizure 10. neuropathologica.119(1):37 9. Neuros neuroscience the: official journal ofInternational the Society for Developmental development and evolution the of vertebrate brain. International journal developmentalof 8. communications.2(1):7. cases of multinodular and vacuolating neuronal tumour.neuropathologica Acta 7. the ILAE Diagnost spectrum of focal cortical dysplasias:consensus a classification proposed by anad hoc ForceTask of SN, Roper Salamon N,Schulze Dubeau F, Duncan J,GuerriniKahane R, P, Mathern G, Najm Barkovich AJ, Battaglia G, Becker A,Cepeda C, Cendes F, Colombo N, Crino CrossP, JH, DelalandeO, 6. neurology.77(4):675 F (2015)Familial focal epilepsy with focal cortical dysplasia dueto DEPDC5 mutations. Annals of Lambrecq V,Vlaicu M, Daniau Bielle M, F, Andermann AndermannE, F,Leguern E, ChassouxF, Picard 5. neuronal tumor of the cerebrum. Neurology.89(3):304 4. lac F, Acampora SimeoneD, A (2000)Synaptic properties neocorticalof neurons inepileptic mice 3. epilepsy expression patterns chlorideof transporters, Na+ 2. Neuroradiology. lesions with the imagingMR appearance of multinodular and vacuolating neu 1. References

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HuseJT,Snuderl JonesM, Brathwaite DT, CD, Altman N, Lavi Saf E, HuseJT,Edgar HallidayM, J Huang XF, H,Zhao Zheng K, Qiu M (2013) Regulation oftiming the of oligodendrocyte Hildebrand Griffin MS, NG, Damiano JA, Cops EJ, Burgess Ozt R, Hevner RF (2015) Brain overgrowth in disorders RTKof Hadjivassiliou G, Martinian SquierL, BlumckeW, I, Aronica Sisodiya E, SM, Thom (2010)M F, Guo Maeda Y, J, Ma Xu J, Horiuchi M, Miers VaccarinoL, F, PleasureD (2010) Pyramidal Garbellide R, BockF, Medici V,RoussetMC, Villani F, Boussadia B, Arango Fukushima S, Yoshida A, NaritaArita Y, H, Ohno M, Miyakita Ichimura Y, ShibuiK, S (2015) EnglundFink C, A, Lau C, Pham D, Daza RA, BulfoneA, KowalczykT, HevnerRF (2005) Pax6, Dougherty MJ, Santi M, Brose MS, C,Ma ResnickAC, SievertAJ, Storm PB, Biegel JA (2010) Larsen Larsen KB, Lutterodt MC, Mollgard K, Moller (2010)M Expression of the homeoboxgenes Lamparello P, Baybis M, PollardEM, J,Hol Eisenstat DD, Aronica E, Crino PB (2007) Komori Arai T, N (2013) Dysembryoplasticneuroepithelial tumor,pure a glial tumor? Kamphuis W, MamberC, Moeton M, Kooijman SluijsL, JA, Jansen AH, Verveer deM, Groot A, Iyer Prabowo A,Anink J, Spliet WG, van Rijen AronicaPC, E (2014) Cell injury and nce the: official journal ofSociety the for Neuroscience.30(36):12036 -

Accepted Article- 30. ial journal the of Histochemistry Society.58(7):669 vascul -

24. eveloping neocortex. The Journal neuroscience of the: official journal of the - 76.

n infocal malformations cortical of development. Brain Pathol.24(1):1 epsy. American journal of human genetics.99(2):423 28.

- , Mikolaenko I, Lavi E, Rosenblum (2013) MK Multinodular and 51. - 68.

studies. Neuropathology : official journal of the Japanese grade neuroepithelial tumor of the young (PLNTY): an

of the cerebrum. Brain tumor pathology.32(2):131 a - like components, aberrant CD34 expression, and - 43.

- PI3K on on cells. Brain : a journal of - AKT signaling: a mosaic of - - grade gliomas. Neuro urk E, Jones NC, Leventer RJ, 78. -

fery R, Sexton 27.

- associated lesion. - - 64. 49. epilepsy. Acta - - 9. Lievano M,

- - Oates A,Oates 29. -

e braine - 6. -

Pathol.26(3):319 Histopathological Study in Normal and Pathological Neocortical Resections in Focal Epilepsy. Brain AW, Eriksso 46. adult late with 45. morphology. Acta neuropathologica.131(6):833 neuroepithelial tumors: BRAF, FGFR1,and MYB mutations atoccur high frequency and align with G, Down H, Perry A, Shurtleff S, Gajjar A,Boop FA, Klimo PD,Jr., Mardis ER, Wilson RK, BakerSJ, Zhang J,Wu Brat D, PeterCollins V, DahiyaS, George D, Konomos KurianW, KM, McFadden K, Serafini LN, Nickols Punchihewa C, Easton J, MulderH, Boggs Shao K, Y, R 44. proteins and caspase Rijen PC, FerrierCH, Thom AronicaM, E (2015) Expression neur of 43. mTOR signaling activation in glioneuronal tumors. Brain Pathol.24(1):52 Rijen PC, FerrierCH, Capper D, Thom AronicaM, E (2014) BRAF V600E mu 42. dysplasias. Journal neuropathologyof and experimental neurology.69(8):850 Crino (2010)PB Early progenitor cell marker expression distinguishestype II from type 41. with a Multinodular and Vacuolating Neuronal Tumor of the Cerebrum: ANew "Leave Alone" Me Lesion Warmuth 40. Neuropathology.35(6):561 neuronal tumors. Neuropathology official: journal of the Japanese Society of (2015)Localized overexpression alphaof 39. pyramidal projection neurons. Neuroscienceresearch.55(2):105 38. neuroscience.8:27. view on the functional regulation theof neuronal K(+) 37. pathologies. Neuropathol Appl Neurobiol.35(4):394 Expression patterns glialof fibrillary acidic protein (GFAP) 36. regulation of expression and contribution to brain tumors. CNS oncology.5(3):159 35. central nervous system. Revised 4th edition. Edition. 34. Tumors theof Central Nervous System:summary. a Actaneur H, WiestlerOD, Kleihues P, Ellison DW (2016) The 2016 World Health Organization Classification of 33. development in epilepsy. Actaneuropathologica communications.2: High 32. 48. unresolved questions from experimental and human brain studies. Acta neuropathologica.131(1):27 31.

- throughput,automated quantification of white matterneurons in mild malformation of cortical

CharacteristicImaging Pattern. AJNR American journal of neuroradiology. Orlova KA, V, Tsai Baybis M, HeuerGG, SisodiyaS, Thom StraussM, K, Aronica E, Storm PB, Nunes Nagaishi Yokoo M, H,NobusawaS, Fujii SugiuraY, Molnar Z, Cheung AF (2006)Towards the classification subpopulations of layerof V Medina I,Friedel RiveraP, C, Kahle KT, Kourdougli N, Uvarov P, Pellegrino C (2014)Current Martinian L, Boer K, Middeldorp J,Hol EM, Sisodiya SM, Squier AronicaW, E, Thom M (2009) S,Mansouri Nejad R, Karabork M, Ekinci C, Solaroglu I, AldapeKD, Zadeh G (2016) Louis DNe, Ohgaki He, WiestlerODe, CaveneeWKe WHOclassification of tumours of the Louis DN, Perry A, ReifenbergerG, von Deimling A,Figarella Li Lewis J,Dickson DW(2016) Propagation of tau pathology: hypotheses,discoveries, and yet Reeves C, Tachrount M, Thomas D, MichalakZ, LiuJ, Ellis M, B,Diehl Miserocchi A, McEvoy Ratilal B, McEvoy A,Siso Qaddoumi I,Orisme W, Wen J,Santiago GuptaT, K, Dalton JD, Tang B, Haupfear K, Prabowo AS, IyerAM, Veersema TJ, AninkJJ, Schouten Prabowo AS, IyerAM, Veersema TJ, AninkJJ, Schouten ing JR, TatevossianEllison RG, DW (2016)Genetic alterations inuncommon low - u JY,Ellis M, Brooke Metz M, AlvesHC, Goncalves FG, Kleinschmidt n S, Yousry T, Thom M (2016)Combined ExVivo 9.4T MRI and Quantitative - RH, Hsu CC, daRocha AJ, Amaral do LLF, Godoy LFS, Watkins TW, Marussi VH, onset refractory epilepsy. Neuropathol Appl Neurobiol.33(6):706 - 33.

Accepted Article

- Ball H,Tisi J, de Eriksson SH, BrandnerS, Sisodiya SM, Thom (2014)M diya S,Thom M, Toma A(2007) cerebralDiffuse gangliocytoma inan - internexin within nodules in multinodular and vacuolating - 45.

- 405.

usch Becksfort M, J, Gupta WangP, S, Lee RP, - Cl(

- - - DeMastersBK, Osborn AG (2017) ) cotransporte) delta i Y, SuzukiTanaka R, Suzuki Y, K, Hyodo A opathologica.131(6):803 - - van Meeteren AY,Spliet WG, van van Meeteren AY,Spliet WG, van - n epilepsy 15. odegenerative disease -

72. Branger CaveneeD, WK, Ohgaki

- r KCC2. Frontiers incellular 66. tation is associated with - associated lesional

- 63. -

9. - 73.

- - I focal cortical 20. e15. - - grade related

Sox2:

- Neurobiol.31(6):580 (2005)Mcm2 labelling balloon of cells in focal cortical dysplasia. Neuropathol Appl 61. dysplasia typeIIB. Journal neuropathologyof and experimental neurology.66(11):1045 SisodiyaSM (2007)An investigation of the expression G1of 60. in grey matter heterotopia in patients with epilepsy. Epilepsia.45(8):916 59. microdysgenesis. Acta neuropathologica.105(6 inhibitoryinterneuronal populations and neuropeptideY expression infocal cortical dysplasia and 58. Pa 57. in the mammalian target of rapamycin regulator NPRL3. Annals of neurology.79(1):132 CrinoMB, BahloPB, Mandelstam SA, McGillivray G, MacGregor D, Kannan MaixnerL, HarveyW, AS, Amor DJ, Delatycki 56. II. type Epilepsia.54(5):898 study whiteof matter hypomyelination and oligodendroglial maturation infocal cortical d 55. tau tangles localize to 54. expression inrefractory human epilepsy. Epilepsyresearch.74(2 53. entity. Brain Pathol. development with oligodendroglial hyperplasia in frontal lobe epilepsy: A new clinico Toledano R, Feucht M, MuhlebnerA, Czech Blumcke T, I (2016) Mild malformation of cortical Bien CG, Polster Schulz T, Kalbhenn R, T, Urbach H, Becker A, Grunwald T, Huppertz 52. sclerosis complex. Brain Pathol. oligodendroglial turnover is associated with myelin pathol HainfellnerJA, Prabowo AS, Reinten RJ,Hoogendijk L, AninkJJ, AronicaE, Feucht (2016)M Impaired 51. extra tumors reveals Reifenberger G, von Deimling A(2011)Analysis BRAFof V600E mutation in 1,320 nervous system Wesseling P, MawrinHasselblatt C, M, Louis DN, Korshunov A, PfisterS, Hartmann Paulus C, W, 50. epileptogeniclesions. Neurobiology disease.95:93of Functional aspects of early brain development arepreserved in tuberous sclerosis complex(TSC) Feucht M, Krsek ZaP, 49. neuroscience.19(1):28 48. neuropatholo (2016)Germline and somatic FGFR1 abnormalities indysembryoplastic neuroepithelial tumors. Acta Zakrzewski K, Sisodiya SM, Paulus W, Albrecht S, Hasselblatt JabadoM, N,Foulkes WD, Majewski J Karamchandani J, Greenwood CM, Berghuis Ellezam B, MichalakZ, Thom M, Lockhart PJ, Leventer RJ,Ohm M, MacGregor D, Jones D, FahiminiyaS, BarekeE, SchullerU, Monoranu CM, Strater R, Kerl K, Niederstadt KurlemannT, G, 47. thol.22(3):350

- cerebellar pilocytic astrocytoma. Acta neuropathologica.121(3):397 Sen A,Thom M, Martinian HardingL, Cross B, JH, Nikolic SisodiyaM, SM (2007) Pathological Sen A,Martinian L, Ni Schurr J, CorasRossler R, K, Pieper T, Kudernatsch M, Holthausen H, Winkler P, Woermann F, Scholl T, Muhlebner A,Ricken G,Gruber V,Fabing A,Samueli S, Groppel G, Dorfer C,Czech T, SchindlerCapper G, D, Meyer RuffoloIyer G, A, Cifelli P, Roseti C, MuhlebnerA, van Scheppingen J,Scholl T, HainfellnerJA, S,Robel SontheimerH (2016)Glia as drivers abnormalof neuronal activity. Nature RiveraGayden B, CarrotT, Thom M, Martinian L, SisodiyaSM, Cross JH, W Thom M, Martinian L, Sen A, SquierHarding W, BN, Cross JH, Harkness Thom M, Martinian L, Parnavelas JG,SisodiyaSM (2004) Distribution of cortical interneurons Thom M, Harding BN,Lin WR, Martinian CrossL, H,SM Sisodiya (2003)Cajal Thom M, BlumckeI, Aronica (2012) E Long Sim JC,Scerri T, Fanjul Shepherd C,LiuJ, GocJ, Martinian Jacques L, Sisodiya TS, SM, Thom M (2013)A quantitative gica.131(6):847

high mutation frequenciesin pleomorphic xanthoastrocytoma, ganglioglioma and - 79.

Accepted Article

- M, M, LockhartPJ, Leventer RJ (2016)Familial cortical dysplasia caused by mutation

8. mecnik J,Jansen FE, Spliet WG, Limatola C, Aronica E, Palma (2016) E

focal cortical dysplasia in older patients. Epilepsia.48(8):1447 -

33.

- 908. This article isprotected by copyright. All rights reserved. - kolic WalkerM, MC, Thom M,SM Sisodiya (2007)Increased NKCC1 63. - Fernandez Riseley M, JR, Gillies G, Pope K, van Roozendaal H, Heng JI,

- Zhang J,Nadaf J,Boshari T, Faur

J, JanzarikW, Omran H, Herold AM, Bens S, Siebert R, Zakrzewska M, Liberski PP, ):561 - term epilepsy - - 9. 101.

illiams G, Stoeber HarknessK, W, Harding BN

ogy infocal cortical dysplasia and tuberous - phasecell cycle proteins infocal cortical - 3):220 - associated tumors. Brain y D,y Zeinieh M, Blanc R, Burk DL, - MendeC, Schmieder K, - 7. - 23. -

405.

W, McEvoy A,

- HJ, Gil 54. - Retzius cells, - pathological - -

55. 7.

ysplasia -

Nagel A, comparisons. Annals of neurology.79(1):42 cortical dysplasia: Exvivo 7Tmagnetic resonanceimaging abnormalitiesand histopathological D'Incerti FreriL, E, Morbin FugnanesiM, Figini V, SpreaficoM, Ga R, 70. neuropathologica.120(1):85 human cortical dysplasia and tuberous sclerosis: Thom M, AndersonHelen G, J, Cross Harkness W, HardingJacques B, TS (2010)Balloon cells in 69. internatio vacuolating neuronal tumor affecting amygdala and hippocampus: Aquasi 68. neuroscience.19(8):1085 (2016)KE Neuronal activity enhances tau propagatio H, Boonen RA, Herman M, Nahmani E, Emrani S,Figueroa YH, Diamond MI, Clelland Wray CL, S, Duff 67. again. Developmental neurobiology.7 66. subventricular neurogenic system specifically express GFAP Bergvan de WD, Huitinga I, EM Hol (2010)Lon 65. transcription factor in brain cancer. Oncotarget. 64. system. Science.351(6271):379 Fancy SP (2016) Oligodendrocyte precursors migrate along vasculaturein developing the nervous 63. theof literature. Journal of neuropathology and experimental neurology. epilepsy series immunohistochemical, with molecular genetic,and clinical correlations and a review Bran SM, 62.

ZuccaI, Milesi G, Medici V, Tassi L, DidatoCardinale G, F,Tringali G, Colombo N, Bramerio M, Yasin SA, Latak BecheriniK, F,Ganapathi A, Miller Campos K, O, Picker SR, Bier N, Smith M, Yamaguchi M, Komori T, NakataY, Yagishita A, Morino IsozakiM, E (2016) Multinodular and Wu JW,Hussaini SA, BastilleIM, Rodriguez GA, Mrejeru A,RilettK, Sanders DW, CookC, Fu Villarroel van denBerge SA, Middeldorp J,Zhang CE, Curtis MA, Leonard BW, Mastroeni D, Voorn P, Tsigelny IF, Tsai HH, Niu J, Munji R, Davalos ChangD, J,Zhang H, Tien AC,Kuo CJ, Chan JR, Daneman R, Thom M, Toma A,An S, Martinian Hadjivassiliou L, G, Ratilal B, Dean A, McEvoy A, Sisodiya dnerS (2011) One hundred anddysembryoplastic one neuroepithelial tumors: an adult nal.66(1):34 - Campos D, Gonzalez

Accepted ArticleKouznetsova VL, Lian N, Kesari S(2016) Molecular mechanisms OLIG2of - 41. - 92.

- 84.

4(10):953 - Billault(2014) C The MAP1B case: an old MAP thatis new - 58. gterm quiescent cells in the aged human

71. isolation pathological a of progenitor

n and tau pathology in vivo. Nature - delta. Aging cell.9(3):313 rbelli R (2016)Type II focal 70(10):859 - tumor? Pathology - 78. - 26. - likecell. Acta