INTRINSIC OR INDUCED AGGRESSIVENESS-LINKED REGULATION OF VEGFC IN MEDULLOBLASTOMA CELL LINES

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Submitted on 4 Dec 2019

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INTRINSIC OR INDUCED

AGGRESSIVENESS-LINKED REGULATION OF VEGFC IN MEDULLOBLASTOMA CELL LINES

Jerome Durivault, Yannick Comoglio, Rita Hanna, Vincent Picco, Alvaro Javier, Feliz Morel, Magalie Leloire, Amandine Morot, Marina Pagnuzzi,

Jérôme Durivault, et al.

To cite this version:

Jerome Durivault, Yannick Comoglio, Rita Hanna, Vincent Picco, Alvaro Javier, et al.. INTRINSIC OR INDUCED AGGRESSIVENESS-LINKED REGULATION OF VEGFC IN MEDULLOBLAS-TOMA CELL LINES. 8ème congrès de la Société Française d’Angiogenèse, Oct 2018, Reims, France. �hal-02382911�

INTRINSIC OR INDUCED AGGRESSIVENESS-LINKED

REGULATION OF VEGFC IN MEDULLOBLASTOMA CELL LINES

Yannick COMOGLIO

a

_{, Rita HANNA}

a

_{, Vincent PICCO}

b

_{, Alvaro Javier FELIZ MOREL}

a

_{, Magalie LELOIRE}

a

_{, Amandine MOROT}

a

_{, Marina PAGNUZZI}

b

_,

Jérôme DURIVAULT

b

_{, Gilles PAGÈS}

ab

_{, Sonia MARTIAL}

a

a : Ins'tute for Research on Cancer and Ageing of Nice (IRCAN), NICE, FRANCE b : Scien'fic Center of Monaco (CSM), MONACO Medulloblastoma (MDB) is the most common pediatric malignant brain tumor. MDB is a very heterogeneous disease that consists of four subgroups, each of them with different molecular profiles, metastasis status and clinical outcomes. Treatment of MDB includes surgery, radiotherapy and chemotherapy. It cures 70% of the pa'ents nevertheless with many side effects. Relapse is always fatal. Understanding the mechanisms of this relapse might lead to the development of new targeted treatments. VEGFC and lympha'c markers are the main actors of the metasta'c process in many tumors. We thus meant to determine their role and regula'on in MDB-derived cells. We demonstrated that low-aggressiveness MDB-derived cells, DAOY, expressed high basal amounts of lympha'c marker mRNAs and proteins (VEGFC, PROX1, NRP2). Conversely, highly aggressive cells, HD-MB03, presented low amounts of these markers. X-ray irradia'on treatment of the cells induced a rise in VEGFC, at the mRNA and protein levels. We thus ques'oned the regula'on of lympha'c markers in MDB cells. We observed that in the highly aggressive HD-MB03 cells, VEGFC mRNA amount relies upon a very ac've NF-κB-dependent promoter, but that the mRNA gets intensely degraded, hence the low level of VEGFC mRNA in these cells. Conversely, DAOY cells present low ac'vity of VEGFC promoter and high stability of the synthesized mRNA. The promoter ac'vity is independent from NF-κB. Moreover, while VEGFC is secreted in DAOY cells, it is retained inside the cytoplasm of HD-MB03 cells. Since VEGFC is so precisely regulated in MDB cells, it may play an important part in MDB aggressiveness.

RegulaOon of VEGFC in medulloblastoma

p VEGFC Renilla p VEGFC Renilla

p VEGFC nFKB mut Renilla Renilla 3’UTR VEGFC

VEGFC is involved in the EMT phenotype of MB cells

MulOple irradiaOons influence MB cell VEGFC producOon and phenotype

N Cad

ARD1

HD-MB03 (Grp3): High promoter acOvity, involvement of NF-κB site

VEGFC mRNA less stable in HD-MB03

Lack of VEGFC secreOon in HD-MB03

HD-MB03 have a higher ability to regulate their VEGFC producOon

Clone1 HD HD VC++

HSP 90

VEGFC

Claudin

DAOY Clone 1 HD-MB03 DAPI DAPI DAPI VEGFC VEGFC VEGFC Merge Merge Merge

Intrinsic characterisOcs of MB cell lines

DAOY (SHH): High expression of lymphaOc markers and VEGFC

> Low proliferaOon and high migraOon rates

HDMB-03 (Grp3): High expression of lymphaOc markers and VEGFC

> High proliferaOon and low migraOon rates

VEGFC has a impact on the proliferaOon of HD-MB03 cells

.

VEGFC DAOY VEGFC promoter VEGFC VEGFC mRNA HD-MB03

Daoy cells (SHH): CRISPR-Cas9 knock-out of VEGFC gene

> Decrease of epithelial phenotype: Claudin, N-cadherin

HD-MB03 (Grp3): TransducOon of VEGFC gene

> Increase the epithelial phenotype : Claudin, N-cadherin

VEGFC increases the mesenchymal-epithelial transiOon of MB cells

HSP90

NFκB VEGFC mRNA DegradaOon VEGFC RetenOon

VEGFC has a key role in MB aggressiveness

VEGFC

Mesenchymal genes

Epithelial genes

Prolifera'on and migra'on

Tube organiza'on

VEGFC is a negaOve regulator of medulloblastoma aggressiveness,

through the mesenchymal-epithelial transiOon

Irradia'on

MulOple irradiaOons influence VEGFC producOon in MB cells

A_er irradiaOon HD-MB03 are more epithelial

MulOple irradiaOons increase VEGFC and the mesenchymal-epithelial transiOon of MB cells

VEGFC producOon Daoy HD-MB03 0 1000 2000 3000 VEGFC (pg/ml/10 6 cells/48 hours) p<0.001 0 2 4 6 8 0 100 200 300 400

Days after seeding

Cell number (fold vs day 1)

Daoy

* *

*

*

*

HD-MB03 VEG FC VEG FD VEG FR3 _NRP2 PROX1 PD PN LYVE1 0 20 40 60 80 Relative mRNA amount Daoy HD-MB03 *** *** *** *

Contact:

yannick.comoglio@unice.fr