Mechanotransductive Activation of b-catenin:

from Mesoderm Evolutionary Emergenceto Cancer Progression

I- Evolutionary implication in mechanotransduction in mesoderm origins, 600 millions years ago

II- From development to cancer: tumour growth mechanical cues in the reactivation of mechanosensitive embryonic pathways

UMR168

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Twist, Snail

Introduction: initiation of morphogenetic movements in Drosophila

embryos

Gènes -> Forces

Costa et al, Development 1993 Supatto et al, PNAS, 2005

Nos, TslBcd, Tsl

Dorsal

Anterior

Length, 500 microns

6000 cells surrounding a yolk

Forces = 10-100 nN

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D. Kiehart, M. Leptin, E. Wieschaus,

N. Perrimon groups et al.

(Adams et al - Pouille, Ahmadi et al, 2009)

Myo-II Apical Stabilisation generates Mesoderm Invagination

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Twi-: stochastic apical constriction

Sna-: neither stochastic nor

collective apical constriction

Stochastic = oscillations

Collective = constricted stabilized

M. Leptin, E. Wieschaus, N. Perrimon et al. Groups Sweeton et al, Development, 1991

A. Martin et al, Nature 2009

Myo-II

There exists two phases of apical constriction:

is the second mechanically triggered by the first?

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Indenting sna mutants to rescue the missing mechanical strains of the

first constricting stochastic phase, rescues the second coordinated

constriction phase and mesoderm invagination

Pouille, Ahmadi et al, Science Signalling, 2009

Demosthène Mitrossilis et al., submitted

Energies Mechano-

Transduction = 10kT

(Forces = pN)

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Second mechanotransductive wave: b-catenin phosphorylation of Y667, impairing

b-catenin interaction with E-cadherin, is mechanically induced by mesoderm

invagination

I- Evolutionary implication of mechanotransductive b-cat

activation in mesoderm emergence, 600 millions years ago

The b-catenin protein: a mechano-genetic dual role

-> b-catenin phosphorylation on its Y654 site leads to b-catenin cytoplasmic

and nuclear translocation:

b-catenin cytoplasmic and nuclear translocation are mechanically induced by

Y667 b-catenin mechanical phosphorylation and lead to Twist expression

maintenance in the mesoderm

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Note: 1%FA Fix procedure for Arm nuclei detection

(poor junctional resolution)

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Zebrafish: the mesoderm patterning gene notail is expressed at the onset

of epiboly in specifically deformed margin cells in a b-catenin dependent

but Wnt independent process

Dkk: inhibitor of Wnts through Lrp6

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The b-catenin dependent expression of notail in margin cells is induced

by the morphogenetic movement of epiboly onset

Uniaxial deformation:

35mm during 20min

-> Magnetic rescue of epiboly initiation in vivo

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Magnetic: 0.25 +- 0.06 mm/minEndogenous: 0.22 +- 0.04 mm/min

Bleb: not detectable (Poky: fluctuations of 1 mm/min)

Ultramagnetic

Liposomes:

Fe3O2 30% (v:v)

Fluo;PEG-tagged

200nm

C. Ménager

(UPMC)

Micromagnets:NdFeB

Embedded in Pdms

BgradB = 10-2 T2/m

at 400mm

Nora Dempsey

(I.Néel, Grenoble)

Nuclear translocation of b-catenin and notail expression are triggered by

mechanical induction of phosphorylation of the Y667- b-catenin at the onset of

epiboly

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Conclusion 1

Mechanical induction of the b-cat/mesoderm pathway at the origin of mesoderm

emergence leading to the Diploblast-Tribloblast evolutionary transition ?

This suggests that mecha-p-Y667 b-cat was a property

of the 600 MY common ancestor, and that it could have

been the primitive inductor of mesoderm emergence

Origin of the mesoderm?

Zebrafish: Nodal -> ntl

Drosophila: Dorsal -> Twist

T. Brunet, Adrien Bouclet et al, Nature Communications, 2013

mecha-p-Y667 b-cat ,

but not Dorsal and Nodal,

is common to the two species

in mesoderm specificationEcdysozoa Vertebrates

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II- From development to cancer: tumour growth mechanical cues in the

reactivation of mechanosensitive embryonic pathways

b-cat dependent mechanical induction of oncogenes expression

In situ ex-vivo: Joanne Whitehead et al, HFSPJ 2008

Control Compressed

c-Myc

(target of b-cat)

Ex vivo mechanical compression of the distal colon

APC1638N/+

PP1 (inhibitor

of Src familly

kinases)

P = 0.8kPa

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Magnetic loading to mimic tumour growth pressure in vivo

Oct. 30th 201515

P= 1.2kPa mean value

(tumour growth pressure: 1kPa)

Magnetic Liposomes

Mechanical induction of oncogenesis by tumour growth pressure

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Endogenous hyperproliferative Notch tumour growth pressure activates the b-cat

tumorogenic pathway in neighbouring non tumour Notch negative crypts

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Conclusion 2- Beta-cat dependent mechanical Induction of Oncogenes

Expression in healthy tissues by tumour growth pressure in vivo

In vivo: Elena Fernandez-Sanchez, Sandrine Barbier et al., Nature, 2015

In APC+/-

“Mechano-Cancer” Consortium

Emmanuel Farge’s team

(UMR168 Institut Curie)

Elena Fernandez-Sanchez

Sandrine Barbier

Anne-Christine Brunet

Adrien Bouclet

Thibaut Brnuet

Joanne Whitehead

Sylvie Robine’s team

(UMR 144 Institut Curie)

Jeanne Netter

Mickael Tanter’s team

(Institut Langevin, ESPCI)

Jean-Luc Gennisson

Heldmuth Latorre-Ossa

Plateforms: Animal

House’s team,

Bioinformatics,

Sequencing

(Institut Curie)

- Isabelle Grandjean

Virginie Dangles-Marie

Stéphanie Boissel

- Elodie Girard

Nicolas Servant

- Thomas Rio-Frio

Christine Ménager’s team

(Université Pierre et Marie Curie Physico-

Chimie, ESPCI) and Sylviane Lesieur’s

team (Université Paris12 Pharma)

Gaelle Bealle

Aude Michel

Hélène Marie

Silvia Fre’s team

(UMR 3215 Institut

Curie)

Mathilde Huygue

Chantal Housset’s team

(Faculté de Médecine, Hopital Saint-Antoine, Inserm)

Colette Rey

Laura Fouassier

Audrey Claperon

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Mechanics & Genetics of

Embryonic and Tumour Development ANR

–Joanne Whitehead, Sandrine Barbier and Elena Fernandez-Sanchez

–Tumoral Progression / Post-docs IC-Marie Curie, ANR, RTRA FPGDG

–Nicolas Desprat

–Ferromagnetic Injections and Mechano-transcription Post-doc INSERM 2005/2008

–Philippe-Alexandre Pouille, Padra Ahmadi,, Démosthène Mitrosslis, Benjamin Driquez, Willy Supatto, A. Bouclet

–Numerical simulations and experiments

–PhD 2005/2009, Microsoft European Grant

–T. Brunet, Adrien Bouclet, L. Henry, F. Serman

–Zebrafish

–Anne-Christine Brunet:

–Ingénieur de recherche

–Collaborators:

Nora Dempsey (I. Néel), C. Ménager, Paris 6) , Emmanuel Beaurepaire(Optics, Polytechnique), Glenn Edwards (Duke U), L. Solnica-Krezel (Zebra)

Silvia Fre, Sylvie Robine (Colon cancer, I. Curie), Michael Tanter (I. Langevin)

ARC, C’Nano, Fondation Pierre Gilles de Gennes, Inca, Labex CelTisPhysBio

Anne Plessis (Droso, IJM, Paris)

Dino Yanicostas (Zebra, UPMC, Paris)

Controls: Ret ligands and UMLs by themself, do not

activate Ret

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