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Thèse De Doctorat Présentée En Vue De L’Obtention Du Grade De Docteur En Interactions"Moléculaire"Et"Cellulaire"

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Thèse De Doctorat Présentée En Vue De L’Obtention Du Grade De Docteur En Interactions École doctorale sciences de la vie et de la santé Unité de recherche : CNRS UMR 7284 INSERM U 1081 UNS Thèse de doctorat Présentée en vue de l’obtention du grade de docteur en Interactions"moléculaire"et"cellulaire" de L’UNIVERSITE COTE D’AZUR par Papa Diogop Ndiaye RÔLE%ET%REGULATION%DU%VEGF0C%DANS%LES% CANCERS%DU%REIN%A%CELLULES%CLAIRES% Dirigée par Dr Gilles Pagés Soutenue le 08 Décembre 2017 Devant le jury composé de : Barbara%Garmy0Susini%%%%%CR1, Université Toulouse Midi-Pyrénées Rapporteur Fabrice Soncin DR2, Université de Lille 1 Science et technologie Rapporteur Gilles Pagés DR1, Université de Nice Sophia Antipolis Directeur de thèse Marc POCARD PuPH, Université Paris Denis-Diderot Examinateur Patrick Auberger DR1, Université de Nice Sophia Antipolis Président RESUME&DE&THESE& ! ! Introduction!:!Les!zones!hypoxiques!sont!des!caractéristiques!communes!des!tumeurs!métastatiques.! Le!carcinome!à!cellules!rénales!(RCC)!exprimant!le!facteur!inductible!de!l'hypoxie!(HIF)!en!raison!de! l'inactivation! du! gène! de! von! Hippel! Lindau! (vhl),! représente! un! modèle! d'hypoxie! chronique.! Le! devenir! des! patients! dépend! du! stade! de! dissémination! des! cellules! tumorales.! Par! conséquent,! déchiffrer! les! mécanismes! de! ! métastase! est! une! préoccupation! majeure.! Le! développement! dépendant! du! VEGFMC! (Vascular! Endothelial! Growth! Factor! C)! d'un! réseau! lymphatique! est! en! première!ligne!de!propagation!métastatique.!! ! Objectifs!:!Démontrer!que!le!VEGFMC!est!régulé!par!l’hypoxie!et!étudier!la!régulation!du!VEGFMC!dans! ce!contexte.!Ensuite!étudier!le!rôle!du!VEGFMC!dans!la!tumorigenèse!des!ccRCCs.! ! Résultats&:! Pour! étudier! le! rôle! de! VEGFC! dans! la! dissémination! du! RCC,! nous! avons! étudié! son! expression! dans! l'hypoxie! et! nous! avons! invalidé! son! gène! dans! des! lignées! cellulaires! modèles! de! souris!humaines!et!murines.!L'hypoxie!régule!négativement!l'ARNm!de!VEGFC!par!une!diminution!de! la!transcription!et!de!la!stabilité!de!l'ARNm!mais!l'expression!de!la!protéine!VEGFMC!est!induite!par! l’hypoxie.! Des! capacités! accrues! de! prolifération! et! de! migration,! une! suractivation! de! la! voie! de! signalisation! AKT! et! une! meilleure! expression! des! marqueurs! mésenchymateux! et! des! marqueurs! souches!caractérisent!les!cellules!vegfMc!M/M.!Alors!que!les!cellules!vegfc!M/M!ne!forment!pas!de!tumeurs! chez! les! souris! immunodéficientes,! elles! développent! des! tumeurs! agressives! chez! les! souris! immunocompétentes.! De! plus,! les! cellules! RCC! de! souris! génèrent! des! tumeurs! à! croissance! rapide! chez!les!souris!invalidées!pour!six1!ou!eya2,!deux!régulateurs!majeurs!de!l'expression!de!VEGFC.!La! surexpression!des!marqueurs!lymphangiogéniques,!y!compris!le!VEGFC,!est!liée!à!une!augmentation! de!la!survie!sans!progression!et!globale!chez!les!patients!atteints!de!tumeurs!non!métastatiques!alors! qu'une! diminution! de! la! survie! sans! progression! et! globale! est! observée! chez! les! patients! métastatiques.!! ! Conclusion!:!Nos!expériences!décrivent!une!régulation!subtile!du!VEGFMC!par!hypoxie!et!mettent!en! évidence!son!rôle!bénéfique!ou!péjoratif.!Par!conséquent,!le!ciblage!VEGFMC!pour!la!thérapie!doit!être! considéré!avec!prudence.! REMERCIEMENTS! Ce"manuscrit"a"pour"but"de"résumer"les"trois"années"que"j’ai"passées"dans"l’équipe"du"Dr" Gilles"Pagès"au"sein"de"L’Institut"de"Recherche"sur"Cancer"et"le"Vieillissement"de"Nice" (IRCAN)."Le"but"du"jeu"est"d’y"mettre"ces"résultats"scientifiques"mais"si"je"devais"retenir" qu’une"chose"de"ces"trois"années"cela"serait"les"rencontres."Il"m’est"donc"impossible"de" finir"cette"thèse"sans"remercier"toutes"les"personnes"rencontrées."" " Mais"tout"d’abord"je"vais"remercier"les"membres"du"jury"qui"ont"accepté"d’évaluer"mon" travail"de"thèse"et"qui"pour"certains"sont"venus"de"très"loin."" " Gilles,"merci"tout"d’abord"de"m’avoir"pris"dans"ton"équipe."Tu"m’as"donné"la"chance"de" pouvoir" effectuer" ma" thèse" malgré" mon" parcours" atypique." L’adaptation" a" été" difficile" pour"moi"en"passant"de"Baharia"à"toi"mais"j’ai"survécu."Avec"toi"j’ai"beaucoup"appris…" de"blagues"sur"les"blondes,"à"ne"pas"manger"avant"midi"et"demi"et"plus"de"choses"qui" resteront" à" jamais" dans" ma" tête." Oui" je" retiens" mieux" les" choses" inutiles." Merci" de" m’avoir" laissé" la" liberté" nécessaire" pour" développer" mon" sujet" de" thèse" et" de" m’avoir" soutenu"durant"tout"ce"temps."Tu"as"été"un"directeur"de"thèse"super"cool"et"aussi"une" belle"personne"qui"prend"soin"des"gens"autour"de"lui."" " Yann,"si"j’ai"fait"cette"thèse"c’est"aussi"grâce"à"toi."Même"si"tu"es"loin,"je"ne"t’oublie"pas." J’ai"beaucoup"apprécié"ta"compagnie."Malgré"le"fait"que"tu"es"une"«"tête"de"con"»,"tu"es"un" vrai"pote."Tu"me"manques."Je"rigole"moins"au"foot"sans"toi."Merci"pour"tout"mec."" " Joff," toi" aussi" tu" as" été" un" acteur" important" dans" mon" recrutement." Je" ne" t’oublie" pas" capitaine."Même"si"tu"es"un"jour"de"niveau"ligue"j’ai"quand"même"joué"avec"toi"et"ça"c’est" cool." Merci" pour" ta" gentillesse" et" tes" conseils" au" labo" comme" au" foot." Dorothée," tu" es" aussi"inclue."" " Julien"Parola,"Sandy,"Maeva"vous"m’avez"intégré"dans"l’équipe"sans"problème."On"a"passé" de"bons"moments"à"la"paillasse"et"aussi"en"dehors"du"laboratoire."Ce"fût"un"plaisir"de" travailler"avec"vous."Les"conditions"de"travail"peuvent"rendre"une"thèse"très"dure"à"vivre" mais"avec"vous,"ces"trois"ans"étaient"aussi"bons"que"du"«"tataki"»."Merci"aussi"pour"votre" aide"dans"le"projet." " Je" remercie" aussi" les" autres" membres" de" l’équipe" Pagès" qui" nous" ont" rejoint":" Sonia" Banana,"Yannick,"Audrey"ma"médecin"préférée,"Jérôme"et"Marilena"qui"n’est"plus"avec" nous"au"labo."Sonia,"tes"chansons"ne"vont"pas"me"manquer"mais"ta"joie"de"vivre"et"ton" sourire"oui."" " Je"n’oublie"pas"toute"l’équipe"Pouysségur"à"commencer"par"Jacques."Je"pense"que"j’ai"de" la"chance"d’avoir"côtoyer"un"scientifique"d’une"telle"qualité."J’ai"beaucoup"appris"à"ses" cotés."Merci"Nathalie"pour"tes"conseils"qui"m’ont"beaucoup"aidé"tout"au"long"de"ma"thèse" et"ton"accessibilité."Philippe"et"Roser,"vous"m’avez"beaucoup"aidé"professionnellement"et" aussi" personnellement." Merci" pour" tout." Cercina," merci" pour" ta" gentillesse" et" tes" conseils";"cela"m’a"beaucoup"aidé."Masa"et"Lucilla,"membre"de"la"famille"Banana"merci" d’avoir"été"là."Même"sans"fenêtre,"j’avais"une"belle"vue."C’est"toujours"bon"pour"le"moral." Je" n’oublie" pas" nos" médecins" Nirvana" et" Julie." Pour" finir," Ibtissam," je" te" remercie" 2" "" beaucoup"pour"ta"compréhension"tes"conseils"ton"soutien"au"labo"et"en"dehors."Tu"es" une"belle"personne."" Je" remercie" aussi" les" stagiaires" qui" sont" passés" par" le" labo" durant" ces" 3" ans" comme" Lorenza," Giulia," Natacha," Alvaro," Monique," Willian" et" une" pensée" spéciale" pour" Janita" avec"qui"j’ai"travaillé"pendant"9"mois"et"qui"m’a"beaucoup"aidé"dans"mon"projet."" " Baharia,"tu"as"joué"un"grand"rôle"dans"mon"parcours"scientifique."Je"te"dois"beaucoup."Tu" as"toujours"été"présente"pour"moi"et"pour"cela"je"ne"pourrais"jamais"te"remercier"assez." Tu"m’as"toujours"soutenu"avant"et"pendant"cette"thèse."Merci"infiniment."" Je"remercie"aussi"toute"l’équipe"Hofman":"Paul"Hofman,"Patrick,"Valerie,"Titou,"Jéremie," Barnabé," Nathalie," Laetitia," Mika," Louis," Simon," Iris," MarieGAngela," Olivia" et" Jonathan" sans" oublier" ceux" qui" sont" partis" " vers" d’autres" aventures" (Amine," Josy" et" Laurie)." Je" remercie" également" les" membres" de" l’équipe" Bulavin" avec" qui" j’ai" partagé" le" labo" pendant"une"partie"de"ma"thèse":"Gwen,"Clément,"Laurent,"Caroline,"Sacha"et"Doria." " Je" remercie" aussi" tous" les" monégasques" du" CSM":" Jérôme" " pour" les" CRISPRs" et" tes" conseils,"Scott,"Vincent,"Valérie,"Milica"et"Renaud."" " Merci"à"tous"mes"partenaires"de"sport"à"commencer"par"Julien"qui"m’a"fait"découvrir"le" Kali" et" qui" est" maintenant" un" ami." Toutes" les" personnes" qui" s’entrainent" avec" nous"maintenant." Les" partenaires" du" foot" aussi," je" vous" remercie" avec" une" mention" spéciale"pour"Manu"sans"oublier"les"membres"du"botafogo."Le"sport"m’a"beaucoup"aidé" dans"cette"aventure."" " Merci" à" toutes" les" personnes" travaillant" au" CAL" ou" à" la" tour" que" j’ai" découvertes" en" travaillant" avec" Baharia" et" durant" ma" thèse" qui" m’ont" aidé." Les" chercheurs" comme" Pascal"Lopez"et"Didier"les"membres"du"personnel"comme"Wiliam,"Sami,"David"et"Michel."" " Merci" à" la" famille" Gouneaud" pour" leur" soutien" inconditionnel," à" toutes" les" belles" personnes"rencontrées"à"Brignoles,"à"mes"amis"Celia,"Amaury,"frank,"Audrey,"Amelie"et" Tous"les"membres"de"la"team"Celia"et"la"famille"Huguette." " Je" vais" finir" par" ma" famille" que" je" ne" vais" pas" remercier" car" il" m’est" impossible" de" la" remercier" pour" ce" qu’elle" a" fait" pour" moi" et" pour" que" je" sois" là" où" j’en" suis." J’ai" les" meilleurs"parents"du"monde"qui"m’ont"donné"la"meilleure"éducation"et"je"passerai"ma" vie"à"essayer"de"les"rendre"fier."Ils"m’ont"donné"trois"magnifiques"sœurs"à"qui"j’essaie"de" donner"le"bon"exemple."Je"suis"fier"de"faire"partie"de"cette"famille."" " La"dernière"mais"pas"la"moindre,"Je"remercie"Almut"Brand"qui"s’est"occupée"de"moi"et" sans"cela"tout"aurait"été"beaucoup"plus"difficile."Je"suis"content"que"tu"sois"dans"ma"vie"et" j’espère" pour" longtemps" encore." Merci" pour" ton" soutien," ton" aide," ta" bienveillance" envers"moi,"sans"oublier"les"autres"Brand"pour"leur"soutien"également."" " " " !! 3" "" REMERCIEMENTS!.............................................................................................................................!2"
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