Deciphering the interac-ons between brain tumors and the surrounding nervous system

Decipheringtheinterac-onsbetween
braintumorsand
thesurroundingnervoussystem
SophiePAUTOT,Ph.D.,
ITAV,Toulouse
31Mai2016,GSO,Toulouse
Openques-ons
Resistance?
Issecretomeenoughforreshaping?
Doesitrequirecell-cellcontact?
Signalingpathwaysinvolved?
Metastasis
Didcancercellsescapethetumor?
Orsurroundingcellmodifierbytumor
S.Pautot,semiraireGSO,31Mai2016
Toimproveourfigh6ngchances
weneedadynamicpictureofthetumorinitssurrounding
=>Invitromodel
S.Pautot,semiraireGSO,31Mai2016
Bridgingthegapbetweeninvivoandinvitro
layered architecture
1014neurons
Organizedcelldistribu-on
-highdensityin3D
-differentcelltype
100um
directedconnec-ons
S.Pautot,semiraireGSO,31Mai2016
Establishinga3Dinvitroculturemodel
20µm
45µm<d
2Ddiv2
S.Pautot,semiraireGSO,31Mai2016
Neuronon3Dsupport
Green=neuron
Red=gliacell
S.Pautot,semiraireGSO,31Mai2016
1-Buildingalayeredarchitecture
S.Pautot,semiraireGSO,31Mai2016
Modular network assembly using colloids
4daysneuronculture
! day0:seededonbeads
! day2:injectDNAofinterest
! day4:beadmixing
-->imaging
S.Pautot,semiraireGSO,31Mai2016
Neuronalnetworkforma-on
2weeksinculture
In vitro model
•  neuron growth and network formation
•  spontaneous activity and plasticity
•  cell migration
Sophie Pautot, Claire Wyart, Ehud Y. Isacoff. “Colloid-guided assembly of
oriented 3D neuronal networks.” Nature Methods 8, p. 735-740 (2008).
S.Pautot,semiraireGSO,31Mai2016
3DE18ratcultureatDIV7
DAPI
SMI
SYNAPSIN
100µm
S.Pautot,semiraireGSO,31Mai2016
3Dfluorescenceimagingofimmunostainedprepara-ons
3DE18atDIV21
An-OGalC
S.Pautot,semiraireGSO,31Mai2016
a.u
.
Neuronalspontaneousac6vitymonitored
withRhod-2acalciumsensi6vedye
a.u.
t (s)
S.Pautot,semiraireGSO,31Mai2016
t (s)
Monitoringac-vityoveralargearea
S.Pautot,semiraireGSO,31Mai2016
How tumor secretome affects the surrounding
neuronal network?
Withorwithout
therapeu-ctreatment
Isolatedcancercells
cancerorganoids
Condi-onedmedium
from
Tumorcells
S.Pautot,semiraireGSO,31Mai2016
Zero
Treated
Zero
Treated
e68
T1
e55
T1
e27
T3
e16
T3
GFAPNes-n
e72
DrugCocktailthatinhibitscancercellsecre-on
e83
Cancercells-treated
CancerCells-NT
Visibledifferencesbetweentreatmentsinlessthan48h
Zero
S.Pautot,semiraireGSO,31Mai2016
Treated
In vitro model for clinical diagnostic and evaluation
of therapeutic treatments
Pa-ent1
Pa-ent2
gem
NT
CTRL
Complementpathologydata
Tosub-typetumorsandtotendtowardadiagnos-c
S.Pautot,semiraireGSO,31Mai2016
Pa-ent3
Does cell-cell contact play a role in
tumor ability to reshape its micro-environment? •  to control 3D cell distribution,
•  to control the injection of tumor cells
•  to visualize changes in tumor cell composition & in
neuronal network cell composition,
•  to assess changes in gene and protein expression
S.Pautot,semiraireGSO,31Mai2016
Goalaims
Pa-entX
-IDrespondingneuronalcellpopula-on
-evalua-onofatreatmentin48h
-previewoftumorinvasivestrategies
Pa-entY
Pa-entW
Integra-onofpa-enthistory
Pa-entZ Pa-entT
-sub-typingoftheknowntumorcategories
=> Help choose rapidly the best course
of action while the patient is still alive!
S.Pautot,semiraireGSO,31Mai2016
Acknowledgements part 2
THANKYOU!
JulieC.
S.Pautot,semiraireGSO,31Mai2016