LRP1, Adipogenesis, ObesityLipolysis AssayCells were incubated in DMEM without serum in the presenceof 2% fatty acid-free bovine serum albumin (A7030, Sigma)overnight. Fresh medium containing isoproterenol was ad<strong>de</strong>d for90 min. Glycerol and NEFA contents in the medium weremeasured with free glycerol reagent (F6428 Sigma) and NEFA Ckit (Wako Chemicals, Germany).References1. Kahn SE, Hull RL, Utzschnei<strong>de</strong>r KM (2006) Mechanisms linking obesity toinsulin resistance and type 2 diab<strong>et</strong>es. Nature 444(7121): 840–846.2. He W, Barak Y, Hevener A, Olson P, Liao D, <strong>et</strong> al. (2003) Adipose-specificperoxisome proliferator-activated receptor gamma knockout causes insulinresistance in fat and liver but not in muscle. Proc Natl Acad Sci U S A 100(26):15712–15717.3. Tontonoz P, Hu E, Graves RA, Budavari AI, Spiegelman BM (1994) mPPARgamma 2: tissue-specific regu<strong>la</strong>tor of an adipocyte enhancer. Genes Dev 8(10):1224–1234.4. Gauthier A, Vassiliou G, Benoist F, McPherson R (2003) Adipocyte low <strong>de</strong>nsitylipoprotein receptor-re<strong>la</strong>ted protein gene expression and function is regu<strong>la</strong>ted byperoxisome proliferator-activated receptor gamma. J Biol Chem 278(14):11945–11953.5. Lillis AP, Van Duyn LB, Murphy-Ullrich JE, Strick<strong>la</strong>nd DK (2008) LDLreceptor-re<strong>la</strong>ted protein 1: unique tissue-specific functions revealed by selectivegene knockout studies. Physiol Rev 88(3): 887–918.6. Beisiegel U, Weber W, Ihrke G, Herz J, Stanley KK (1989) The LDL-receptorre<strong>la</strong>tedprotein, LRP, is an apolipoprotein E-binding protein. Nature 341(6238):162–164.7. Kowal RC, Herz J, Goldstein JL, Esser V, Brown MS (1989) Low <strong>de</strong>nsitylipoprotein receptor-re<strong>la</strong>ted protein mediates uptake of cholesteryl esters <strong>de</strong>rivedfrom apoprotein E-enriched lipoproteins. Proc Natl Acad Sci U S A 86(15):5810–5814.8. Beisiegel U, Weber W, Bengtsson-Olivecrona G (1991) Lipoprotein lipaseenhances the binding of chylo<strong>micro</strong>ns to low <strong>de</strong>nsity lipoprotein receptor-re<strong>la</strong>tedprotein. Proc Natl Acad Sci U S A 88(19): 8342–8346.9. Kounnas MZ, Chappell DA, Wong H, Argraves WS, Strick<strong>la</strong>nd DK (1995) Thecellu<strong>la</strong>r internalization and <strong>de</strong>gradation of hepatic lipase is mediated by low<strong>de</strong>nsity lipoprotein receptor-re<strong>la</strong>ted protein and requires cell surface proteoglycans.J Biol Chem 270(16): 9307–9312.10. Descamps O, Bilheimer D, Herz J (1993) Insulin stimu<strong>la</strong>tes receptor-mediate<strong>du</strong>ptake of apoE-enriched lipoproteins and activated alpha 2-macroglobulin inadipocytes. J Biol Chem 268(2): 974–981.11. Vassiliou G, Benoist F, Lau P, Kavas<strong>la</strong>r GN, McPherson R (2001) The low<strong>de</strong>nsity lipoprotein receptor-re<strong>la</strong>ted protein contributes to selective uptake ofhigh <strong>de</strong>nsity lipoprotein cholesteryl esters by SW872 liposarcoma cells andprimary human adipocytes. J Biol Chem 276(52): 48823–48830.12. Hofmann SM, Zhou L, Perez-Tilve D, Greer T, Grant E, <strong>et</strong> al. (2007) AdipocyteLDL receptor-re<strong>la</strong>ted protein-1 expression mo<strong>du</strong><strong>la</strong>tes postprandial lipidtransport and glucose homeostasis in mice. J Clin Invest 117(11): 3271–3282.13. Neese RA, Misell LM, Turner S, Chu A, Kim J, <strong>et</strong> al. (2002) Measurement invivo of proliferation rates of slow turnover cells by 2H2O <strong>la</strong>beling of the<strong>de</strong>oxyribose moi<strong>et</strong>y of DNA. Proc Natl Acad Sci U S A 99(24): 15345–15350.14. Spiegelman BM, Farmer SR (1982) Decreases in tubulin and actin geneexpression prior to morphological differentiation of 3T3 adipocytes. Cell 29(1):53–60.15. Bour S, Daviaud D, Gres S, Lefort C, Prevot D, <strong>et</strong> al. (2007) Adipogenesisre<strong>la</strong>tedincrease of semicarbazi<strong>de</strong>-sensitive amine oxidase and monoamineoxidase in human adipocytes. Biochimie 89(8): 916–925.16. Zhang H, Links PH, Ngsee JK, Tran K, Cui Z, <strong>et</strong> al. (2004) Localization of low<strong>de</strong>nsity lipoprotein receptor-re<strong>la</strong>ted protein 1 to caveo<strong>la</strong>e in 3T3-L1 adipocytesin response to insulin treatment. J Biol Chem 279(3): 2221–2230.17. Willnow TE, Herz J (1994) Gen<strong>et</strong>ic <strong>de</strong>ficiency in low <strong>de</strong>nsity lipoproteinreceptor-re<strong>la</strong>ted protein confers cellu<strong>la</strong>r resistance to Pseudomonas exotoxin A.Evi<strong>de</strong>nce that this protein is required for uptake and <strong>de</strong>gradation of multipleligands. J Cell Sci 107 (Pt 3): 719–726.18. Zurhove K, Nakajima C, Herz J, Bock HH, May P (2008) Gamma-secr<strong>et</strong>aselimits the inf<strong>la</strong>mmatory response through the processing of LRP1. Sci Signal1(47): ra15.Statistical analysisResults are expressed as means 6 SEM. Statistical differencesb<strong>et</strong>ween two groups were evaluated using Stu<strong>de</strong>nt’s t tests. Thelevel of significance was s<strong>et</strong> at P,0.05.Author ContributionsConceived and <strong>de</strong>signed the experiments: OM ELC. Performed theexperiments: OM CD AM DD. Analyzed the data: OM CC CM PV ELC.Contributed reagents/materials/analysis tools: CC DQ CM PV. Wrote thepaper: OM ELC.19. Prawitt J, Niemeier A, Kassem M, Beisiegel U, Heeren J (2008) Characterizationof lipid m<strong>et</strong>abolism in insulin-sensitive adipocytes differentiated from immortalizedhuman mesenchymal stem cells. Exp Cell Res 314(4): 814–824.20. Chappell DA, Fry GL, Waknitz MA, Iverius PH, Williams SE, <strong>et</strong> al. (1992) Thelow <strong>de</strong>nsity lipoprotein receptor-re<strong>la</strong>ted protein/alpha 2-macroglobulin receptorbinds and mediates catabolism of bovine milk lipoprotein lipase. J Biol Chem267(36): 25764–25767.21. Willnow TE, Goldstein JL, Orth K, Brown MS, Herz J (1992) Low <strong>de</strong>nsitylipoprotein receptor-re<strong>la</strong>ted protein and gp330 bind simi<strong>la</strong>r ligands, includingp<strong>la</strong>sminogen activator-inhibitor complexes and <strong>la</strong>ctoferrin, an inhibitor ofchylo<strong>micro</strong>n remnant clearance. J Biol Chem 267(36): 26172–26180.22. Ji ZS, Brecht WJ, Miranda RD, Hussain MM, Innerarity TL, <strong>et</strong> al. (1993) Roleof heparan sulfate proteoglycans in the binding and uptake of apolipoprotein E-enriched remnant lipoproteins by cultured cells. J Biol Chem 268(14):10160–10167.23. Chappell DA, Fry GL, Waknitz MA, Muhonen LE, P<strong>la</strong>d<strong>et</strong> MW, <strong>et</strong> al. (1993)Lipoprotein lipase in<strong>du</strong>ces catabolism of normal triglyceri<strong>de</strong>-rich lipoproteins viathe low <strong>de</strong>nsity lipoprotein receptor-re<strong>la</strong>ted protein/alpha 2-macroglobulinreceptor in vitro. A process facilitated by cell-surface proteoglycans. J Biol Chem268(19): 14168–14175.24. Heeren J, Niemeier A, Merkel M, Beisiegel U (2002) Endothelial-<strong>de</strong>rivedlipoprotein lipase is bound to postprandial triglyceri<strong>de</strong>-rich lipoproteins andmediates their hepatic clearance in vivo. J Mol Med 80(9): 576–584.25. Huang ZH, Reardon CA, Mazzone T (2006) Endogenous ApoE expressionmo<strong>du</strong><strong>la</strong>tes adipocyte triglyceri<strong>de</strong> content and turnover. Diab<strong>et</strong>es 55(12):3394–3402.26. Yue L, Rasouli N, Ranganathan G, Kern PA, Mazzone T (2004) Divergenteffects of peroxisome proliferator-activated receptor gamma agonists and tumornecrosis factor alpha on adipocyte ApoE expression. J Biol Chem 279(46):47626–47632.27. Gao J, Katagiri H, Ishigaki Y, Yamada T, Ogihara T, <strong>et</strong> al. (2007) Involvementof apolipoprotein E in excess fat accumu<strong>la</strong>tion and insulin resistance. Diab<strong>et</strong>es56(1): 24–33.28. Chappell DA, Inoue I, Fry GL, P<strong>la</strong>d<strong>et</strong> MW, Bowen SL, <strong>et</strong> al. (1994) Cellu<strong>la</strong>rcatabolism of normal very low <strong>de</strong>nsity lipoproteins via the low <strong>de</strong>nsitylipoprotein receptor-re<strong>la</strong>ted protein/alpha 2-macroglobulin receptor is in<strong>du</strong>cedby the C-terminal domain of lipoprotein lipase. J Biol Chem 269(27):18001–18006.29. Nykjaer A, Bengtsson-Olivecrona G, Lookene A, Moestrup SK, P<strong>et</strong>ersen CM,<strong>et</strong> al. (1993) The alpha 2-macroglobulin receptor/low <strong>de</strong>nsity lipoproteinreceptor-re<strong>la</strong>ted protein binds lipoprotein lipase and b<strong>et</strong>a-migrating very low<strong>de</strong>nsity lipoprotein associated with the lipase. J Biol Chem 268(20):15048–15055.30. Terrand J, Bruban V, Zhou L, Gong W, El Asmar Z, <strong>et</strong> al. (2009) LRP1Controls Intracellu<strong>la</strong>r Cholesterol Storage and Fatty Acid Synthesis throughMo<strong>du</strong><strong>la</strong>tion of Wnt Signaling. J Biol Chem 284(1): 381–388.31. Daviaud D, Boucher J, Gesta S, Dray C, Guigne C, <strong>et</strong> al. (2006) TNFalpha upregu<strong>la</strong>tesapelin expression in human and mouse adipose tissue. Faseb J 20(9):1528–1530.32. Fears CY, Grammer JR, Stewart JE, Jr., Annis DS, Mosher DF, <strong>et</strong> al. (2005)Low-<strong>de</strong>nsity lipoprotein receptor-re<strong>la</strong>ted protein contributes to the antiangiogenicactivity of thrombospondin-2 in a murine glioma mo<strong>de</strong>l. <strong>Cancer</strong> Res65(20): 9338–9346.33. Ramirez-Zacarias JL, Castro-Munozledo F, Kuri-Harcuch W (1992) Quantitationof adipose conversion and triglyceri<strong>de</strong>s by staining intracytop<strong>la</strong>smic lipidswith Oil red O. Histochemistry 97(6): 493–497.34. Vignon F, Capony F, Chambon M, Freiss G, Garcia M, <strong>et</strong> al. (1986) Autocrinegrowth stimu<strong>la</strong>tion of the MCF 7 breast cancer cells by the estrogen-regu<strong>la</strong>ted 52K protein. Endocrinology 118: 1537–1545.PLoS ONE | www.plosone.org 10 October 2009 | Volume 4 | Issue 10 | e7422
D. CONCLUSION ETPERSPECTIVES67
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Université Montpellier I UFR Méd
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Un grand merci à tous mes collabor
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TitleBreast cancer and tumoral micr
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Cancer du sein et micro-environneme
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C. PRESENTATION DU TRAVAIL DE THESE
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But de la thèseLa cathepsine-D (ca
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I. Le tissu adipeux1) Généralité
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(Adenosine triphosphate). Cette pro
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Cette synthèse de novo a lieu dans
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ABFigure 6 : Schéma de la oxydati
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d. Ladipocyte : une cellule sécré
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3) Ladipogenèsea. Les différentes
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. Les facteurs de transcriptionLa d
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Lexpression de C/EBP est quant à e
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adipeux. Les analyses histologiques
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adipocyteFigure 10 : Schéma repré
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II. La cathepsine-D1) Synthèse, ma
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les cath-B et L, en une forme matur
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Il existe deux RM6P : le RM6P/IGFII
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2) Fonctions de la cath-Da. Dans la
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. Dans les pathologiesEn plus de se
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c. Rôles et mécanismes daction da
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carcinome de prostate serait respon
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La cath-D joue donc un rôle import
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Une fois le marquage des peptides e
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Afin de réaliser ce projet, nous a
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III. Le récepteur LRP11) Organisat
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- Page 58 and 59: LRP1αLRP1Membrane associatedprotea
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- Page 62 and 63: I. Etude du rôle de la cathepsine
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- Page 66 and 67: IntroductionConsumption of meals ri
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- Page 72 and 73: DiscussionOur results demonstrate t
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- Page 86 and 87: ABcath-D mRNA(ratio RS9)PPARg mRNA(
- Page 88 and 89: A D3 D7 D14D0BaD0 D3 D7 D14D0 D3 D7
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- Page 96 and 97: 2) Article 2:LRP1 receptor controls
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- Page 100 and 101: LRP1, Adipogenesis, ObesityFigure 3
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- Page 104 and 105: LRP1, Adipogenesis, ObesitySome ins
- Page 108 and 109: CONCLUSIONLa cathepsine D (cath-D)
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- Page 116 and 117: REFERENCESAhima, R. S. (2006). Adip
- Page 118 and 119: implicates them as antigen presenti
- Page 120 and 121: Cataldo, A. M., Barnett, J. L., Ber
- Page 122 and 123: Folkman, J. (2003). Fundamental con
- Page 124 and 125: Hofmann, S. M., Zhou, L., Perez-Til
- Page 126 and 127: Langin, D. (2006a). Adipose tissue
- Page 128 and 129: Ludwig, T., Ovitt, C. E., Bauer, U.
- Page 130 and 131: Nirde, P., Derocq, D., Maynadier, M
- Page 132 and 133: Rozanov, D. V., Hahn-Dantona, E., S
- Page 134 and 135: Taleb, S., Cancello, R., Clement, K
- Page 136 and 137: Xiao, Y., Junfeng, H., Tianhong, L.
- Page 138 and 139: F. ANNEXE98
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- Page 142 and 143: INTRODUCTIONLysosomal aspartic prot
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REFERENCES1. Vignon, F., Capony, F.
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steps in vivo: proliferation, angio
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28. Laurent-Matha, V., Lucas, A., H
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42. Zurhove, K., Nakajima, C., Herz
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Figure 1. Cath-D interacts with the
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Figure 2. Cath-D binds to residues
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Figure 3. Cath-D interacts with LRP
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Figure 5. Silencing LRP1 in cath-D
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Figure 1. Cath-D interacts with the
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Figure 6. LRP1 is the receptor medi
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Beaujouin, Figure Sup. 2cath-D-/-ME
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RésuméLaspartyl protéase catheps