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, Raffaella Soleti 1 , Luisa Vergori 1 , Grégory Hilairet 1 , Morgane Le Mao 2 , Naig Gueguen 2 , Salim Khiati 2 , Ludovic Clarion 4 , Norbert Bakalara 5 , Stéphane Germain 3* , Guy Lenaers 2* , Ramaroson Andriantsitohaina 1 ?* 1. INSERM UMR1063, Stress Oxydant et Pathologies Métaboliques
, INSERM U 1083, vol.6015
, SAS, p.34090
* Co-last authors ? Corresponding author: Ramaroson Andriantsitohaina, INSERM UMR1063, Stress Oxydant et Pathologies Métaboliques, p.49933 ,
Phone: +33 2 44 68 85 80. E-mail: ramaroson.andriantsitohaina@univ-angers.fr Running Title: Anti-angiogenic properties of PST 3.1a. added at 4 °C overnight to allow antibody interaction with VEGFR2 or Gal-1 protein ,
, Subsequently, the tubes containing the immunoprecipitates were centrifuged, and the supernatants were eliminated. Pellets were washed two times with 100 µL lysis buffer diluted to 1:5 with PBS
, Gel-loading bugger (15 µL) was then added to the pellets. The tubes were heated for 10 min at 70
, The gel was trans-blotted to perform Western analysis for VEGFR2 and Gal-1 proteins as described above. Data analysis Data are represented as mean ± SEM, °C, after which proteins were separated on 4-12% SDS-PAGE gel
, Statistical analyses were performed either using one-way ANOVA (parametric) for all in vitro angiogenesis experiments, or using Mann-Whitney U-tests (non-parametric) for other experiments, using Prism software package 5.00 (GraphPAD Software, CA)
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, Suppl Fig 2: PST 3.1a does not disrupt zebrafish heartbeats
dsRed) eggs were exposed or not to PST 3.1a at 50, 72 and 100 µM for 72, 96 or 120 hpf. A. Representative pictures showing Zebrafish embryos heartbeats (see Video Suppl Fig. 2 for corresponding videos) in the absence or in the presence of PST 3.1a. B. Quantitative measurement of zebrafish heartbeats was reported as number of heartbeats per min ,
, Lors de cette étude, nous avons dans un premier temps caractérisé les propriétés anti-angiogéniques de PST 3.1a, in vitro puis in vivo. Nous avons également démontré que PST 3.1a réduit la vascularisation tumorale et la progression du GBM. Dans un second temps, nous nous sommes intéressés aux perturbations métaboliques endothéliales mises en jeu. Nos résultats prouvent que PST 3.1a réduit l'expression de PFKFB3, enzyme clé pour réguler la glycolyse et la différentiation des CEs lors de l'angiogenèse. De plus, Ce travail de thèse a donc porté sur l'implication du glycomimétique PST 3.1a sur l'angiogenèse physiologique et tumorale, ainsi que sur le métabolisme endothélial associé
,
est une étape essentielle pour la croissance tumorale, l'invasivité et la dissémination métastatique. Bien qu'ils soient principalement utilisés en tant qu'adjuvants à la chimio-et radiothérapie, les traitements anti-angiogéniques représentent pourtant un enjeu majeur dans le cadre des thérapies anti-tumorales. Parmi les tumeurs, le GBM est caractérisé d'un point de vue physiopathologique par son caractère très invasif et hautement angiogénique, L'excès d'angiogenèse est dorénavant bien décrit comme étant impliqué dans la genèse L'angiogenèse ,
2017) font état d'une activité anti-tumorale in vivo de PST 3.1a et nos résultats le confirment. De façon intéressante, PST 3.1a n'induit pas de mortalité ni de perte de poids chez les souris traitées avec 15 mg/kg pendant 14 jours. La même observation est effectuée à très forte dose chez les souris (900 mg/kg pendant 28 jours, non publié), ainsi que chez le zebrafish aux doses utilisées (100 µM pendant 5 jours). Dans cette étude, nous démontrons en plus que PST 3.1a réduit fortement l'angiogenèse tumorale, de presque 50 %. Même si nos résultats ne représentent pas l'effet de PST 3.1a sur les autres modes de vascularisation du GBM, l'angiogenèse représente le principal moteur de la vascularisation du GBM et PST 3.1a la réduit de manière significative. Ces résultats sont particulièrement intéressants car le microenvironnement vasculaire tumoral est très différent du microenvironnement vasculaire physiologique ; les vaisseaux tumoraux sont décrits comme tortueux, instables, peu perméables, hétérogènes, et les CEs qui les composent possèdent de grandes et lâches jonctions inter-cellulaires, les traitements anti-angiogéniques actuels reposent principalement sur l'utilisation du bevacizumab, mais il provoque une forte toxicité chez les patients, 2011. ,
Une seconde importante limite à l'utilisation du bevacizumab concerne son efficacité nouveau paradigme anti-angiogénique, 2015. ,
Ces deux types cellulaires sont décrits comme « brothers in arms, et possèdent plusieurs similitudes métaboliques. Le blocage de ces voies métaboliques communes est stratégique pour les futures thérapies antique l'angiopoïetine like 4, 2006. ,
Comme décrit précédemment, un des principaux mécanismes de résistance tumorale vise à augmenter l'interaction VEGFR2 et Gal-1. Afin de contrer les thérapies actuelles, certaines tumeurs mettent en place un autre mécanisme de résistance, en stimulant la sécrétion d'autres facteurs pro-angiogéniques et notamment le FGF, pour favoriser la vascularisation par d'autres voies indépendantes, Une seconde perspective potentielle viserait à étudier l'impact de PST 3.1a sur la voie du FGF, 2004. ,
une dernière perspective conduirait à étudier l'implication de PST 3.1a sur d'autres pathologies caractérisées par une angiogenèse excessive (Carmeliet, 2003. ,
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