Generation of a genetically engineered total PPARg null mouse line, a new model to study the function of the receptor  Page description

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Details of project

  
Identifier
100196
Type K
Principal investigator Nagy, László
Title in Hungarian Egy uj PPARg null egértörzs létrehozása, a receptor működésének tanulmányozására
Title in English Generation of a genetically engineered total PPARg null mouse line, a new model to study the function of the receptor
Keywords in Hungarian zsírsejt differenciálódás, PPARg, transzkripció, metabolizmus, immunfunkcio
Keywords in English fat cell differentiation, PPARg, transcription, metabolic regulation, immunity
Discipline
Biological basis of diseases related to the above (Council of Medical and Biological Sciences)40 %
Organ physiology and pathophysiology (Council of Medical and Biological Sciences)40 %
General biochemistry and metabolism (Council of Medical and Biological Sciences)20 %
Ortelius classification: Molecular markers and recognition
Panel Physiology, Pathophysiology, Pharmacology and Endocrinology
Department or equivalent Department of Biochemistry and Molecular Biology (University of Debrecen)
Participants Dániel, Bence
Dezső, Balázs
Pap, Attila
Simándi, Zoltán
Varga, Tamás
Starting date 2012-01-01
Closing date 2015-12-31
Funding (in million HUF) 40.000
FTE (full time equivalent) 7.90
state closed project
Summary in Hungarian
A zsírsavak által szabályozott transzkripciós faktor, PPARg szerepe ismert a zsírsejtek differenciálódásában, a makrofágok és dendritikus sejtek funkcióiban és bizonyos betegségekhez is kapcsolódik a receptor mint a cukorbetegség, érelmeszesedés és krónikus gyulladások. Ugyanakkor a receptornak egy adott sejtben betöltött szerepe nehezen tisztázható genetikai modellek nélkül, amelyekben a receptor teljesen hiányzik. Ebben a pályázatban egy olyan egértörzset kívánunk létrehozni, amelynek minden sejtjéből hiányzik a receptor. Ezenkívül olyan kiméra egyedeket is létrehozunk, melyeknek csontvelői sejtjei PPARg -/- genotípusúak, de az állat többi sejtje vad tipusú, vagy a csontvelői sejtek vad típusúak és az állat többi sejte -/- genotípusú PPARg-ra nézve. In vitro kísérletes célokra embrionális fibroblasztokat izolálunk az így létrehozott új egértörzsekből, amelyekből aztán indukált pluripotens őssejteket készítünk transzpozon alapú génbevitel által. Az így létrehozott állat- illetve sejtes modellek lehetőséget biztosítanak, hogy a receptor funkcióját tisztázzuk in vivo illetve differenciálódási modellekben. Ezek a kísérletek segíthetnek megérteni a receptor integráló funkcióját a lipid anyagcsere és az immunrendszer vonatkozásában és in vivo es in vitro betegségmodellek kialakítását is lehetővé teszik.
Summary
The fatty acid activated transcription factor PPARg has been linked to adipocyte differentiation, macrophage and dendritic cell function and also to diseases such as diabetes, atherosclerosis and chronic inflammation. However assigning cellular functions to the receptor in various cell types has been hampered by the lack of appropriate genetic models in which PPARg is completely eliminated from a given cell type or an entire animal. In this proposal we aim at the development and characterization of a complete PPARg null mouse line and also bone marrow chimeras in which bone marrow derived cells are either lacking PPARg in the context of a wild type mouse or have a wild type bone marrow in the context of a PPARg null animal. We will also generate PPARg +/+, +/- and -/- embryonic fibroblasts and induced pluripotent stem cells from these mice to allow investigations of the cellular role of this transcription factor in order to clarify the receptors role in physiological and pathophysiological processes and lead to the development of disease models in vivo and in vitro.




 

Final report

  
Results in Hungarian
A zsírsavak által szabályozott transzkripciós faktor, PPARg szerepe ismert a zsírsejtek differenciálódásában, a makrofágok és dendritikus sejtek funkcióiban, valamint olyan betegségekben is szerepet játszik, mint a cukorbetegség, érelmeszesedés és krónikus gyulladások. Ugyanakkor a receptornak egy adott sejtben betöltött szerepe nehezen tisztázható genetikai modellek nélkül, amelyekben a receptor teljesen hiányzik. Ebben a pályázatban egy olyan egértörzset hoztunk létre, amelynek minden sejtjéből hiányzik a receptor. Elvégeztük a PPARg hiányos egerek metabolikus, hisztológiai és génexpressziós karakterizálását. Csontvelő transzplantációs módszerrel olyan kiméra egyedeket is létrehozunk, melyeknek csontvelői sejtjei PPARg -/- genotípusúak, de az állat többi sejtje vad típusú. A kiméra egerekből csontvelői eredetű makrofágokat differenciáltattunk és RNA-Seq módszerrel vizsgáltuk a PPARg hiányában létrejövő globális génexpressziós változásokat. In vitro kísérletes célokra a PPARg hiányos egerekből immortalizált csontvelői eredetű makrofág sejtvonalat is létrehoztunk, valamint embrionális fibroblasztokat izoláltunk, amelyekből aztán indukált pluripotens őssejteket készítettünk és vizsgáltuk a PPARg szerepét a zsírsejt differenciálódásban. Ezek a kísérletek segíthetnek megérteni a receptor integráló funkcióját a lipid anyagcsere és az immunrendszer vonatkozásában, valamint in vivo és in vitro betegségmodellek kialakítását is lehetővé teszik.
Results in English
The fatty acid activated transcription factor PPARg has been linked to adipocyte differentiation, macrophage and dendritic cell function and also to diseases such as diabetes, atherosclerosis and chronic inflammation. However assigning cellular functions to the receptor in various cell types has been hampered by the lack of appropriate genetic models in which PPARg is completely eliminated from a given cell type or an entire animal. We generated a complete PPARg null mouse line and characterized the metabolic, histologic and gene expression features of these mice. We generated bone marrow chimeras in which bone marrow derived cells are lacking PPARg in the context of a wild type mouse. We differentiated bone marrow derived macrophages from these chimeras and investigated the global gene expression changes by RNA-Seq method. We also generated PPARg null immortalized bone marrow-derived macrophage cell line for in vitro experiments. We isolated embryonic fibroblasts from these mice and generated induced pluripotent stem cells to investigate the cellular role of this transcription factor during adipocyte differentiation. Our experiments help to clarify the role of this receptor in context of lipid metabolism and immune functions and lead to the development of disease models in vivo and in vitro.
Full text https://www.otka-palyazat.hu/download.php?type=zarobeszamolo&projektid=100196
Decision
Yes





 

List of publications

  
Czimmerer Z, Varga T, Kiss M, Ovando Vázquez C, Doan-Xuan QM, Rückerl D, Tattikota SG, Yan X, S Nagy Z, Daniel B, Poliska S, Horvath A, Nagy G, Varallyay E, Poy MN, Allen JA, Bacso Z, Abreu-Goodger C, Nagy L: The IL4-STAT6 signaling axis establishes a conserved microRNA signature in human and mouse macrophages regulating cell survival via miR-342-3p, GENOME MED 8: (1) 63, 2016
Simandi Z, Horvath A, Nagy G P, Nagy L: Prediction and Validation of Gene Regulatory Elements Activated During Retinoic Acid Induced Embryonic Stem Cell Differentiation, JOVE-J VIS EXP : Paper e53978. - közlésre elfogadva, 2016
Tamas Varga, Rémi Mounier, Attila Horváth, Sylvain Cuvellier, Florent Dumont, Szilárd Póliska, Hamida Ardjoune, Gaëtan Juban, László Nagy, Bénédicte Chazaud: Highly dynamic transcriptional signature of distinct macrophage subsets during sterile inflammation, resolution, and tissue repair, J IMMUNOL *: (*), 2016
Ixchelt Cuaranta-Monroy, Zoltan Simandi, Laszlo Nagy: Differentiation of Adipocytes in Monolayer from Mouse Embryonic Stem Cells, METHODS MOL BIOL 1341: 407-415, 2015
Simándi Zoltán, Czipa Erik, Horváth Attila, Kőszeghy Áron, Bordás Csilla, Póliska Szilárd, Juhász István, Imre László, Szabó Gábor, Dezső Balázs, Barta Endre, Sauer Sascha, Károlyi Katalin, Kovács Ilona, Hutóczky Gábor, Bognár László, Klekner Álmos, Szűcs Péter, Bálint Bálint László, Nagy László: PRMT1 and PRMT8 regulate retinoic acid-dependent neuronal differentiation with implications to neuropathology, STEM CELLS 33: (3) 726-741, 2015
Brazda P, Krieger J, Daniel B, Jonas D, Szekeres T, Langowski J, Tóth K, Nagy L, Vámosi G: Ligand binding shifts highly mobile RXR to chromatin-bound state in a coactivator-dependent manner as revealed by single cell imaging., MOL CELL BIOL 34: (7) 1234-1245, 2014
Daniel B, Nagy G, Hah H, Horvath A, Czimmerer Z, Poliska P, Gyuris T, Keirsse J, Gysemans C, Ginderachter J, Balint B L, Evans R M, Barta E, Nagy L: The Active Enhancer Network Operated by Liganded RXR Supports Angiogenic Activity in Macrophages, GENE DEV 28: (14) 1562-1577, 2014
Dozsa A, Dezso B, Toth BI, Bacsi A, Poliska S, Camera E, Picardo M, Zouboulis CC, Biro T, Schmitz G, Liebisch G, Ruhl R, Remenyik E, Nagy L: PPARgamma-Mediated and Arachidonic Acid-Dependent Signaling is Involved in Differentiation and Lipid Production of Human Sebocytes., J INVEST DERMATOL 134: (4) 910-920, 2014
Kotlinowski J, Grochot-Przeczek A, Taha H, Kozakowska M, Pilecki B, Skrzypek K, Bartelik A, Derlacz R, Horrevoets A J G, Pap A, Nagy L, Dulak J, Jozkowicz A: PPARγ activation but not PPARγ haplodeficiency affects proangiogenic potential of endothelial cells and bone marrow-derived progenitors, CARDIOVASC DIABETOL 13: 150, 2014
Brignull LM, Czimmerer Z, Saidi H, Daniel B, Villela I, Bartlett NW, Johnston SL, Meira LB, Nagy L, Nohturfft A: Reprogramming of lysosomal gene expression by interleukin-4 and Stat6., BMC GENOMICS 14:, 2013
Gyongyosi A, Szatmari I, Pap A, Dezso B, Pos Z, Szeles L, Varga T, Nagy L: RDH10, RALDH2 and CRABP2 are required components of PPARg-directed all-trans-retinoic acid synthesis and signaling in human dendritic cells, J LIPID RES 54: (9) 2458-2474, 2013
Mesko B, Poliska S, Váncsa A, Szekanecz Z, Palatka K, Hollo Z, Horvath A, Steiner L, Zahuczky G, Podani J, Nagy L: Peripheral blood derived gene panels predict response to infliximab in rheumatoid arthritis and Crohn’s disease, GENOME MED 5:, 2013
Nagy Zs, Czimmerer Zs, Nagy L: Pro-inflammatory cytokines negatively regulate PPARg mediated gene expression in both human and murine macrophages via multiple mechanisms, IMMUNOBIOLOGY 218: (11) 1336-1344, 2013
Varga T, Mounier R, Gogolak P, Poliska S, Chazaud B, Nagy L: Tissue LyC6- macrophages are generated in the absence of circulating LyC6- monocytes and Nur77 in a model of muscle regeneration., J IMMUNOL 191: (11) 5695-5701, 2013
Czimmerer Z, Varga T, Poliska S, Nemet I, Szanto A, Nagy L: Identification of novel markers of human alternative macrophage activation including potential endogenous PPARγ ligand production mechanisms, IMMUNOBIOLOGY 217: (12) 1301-1314, 2012
Nagy, L., Szanto, A., Szatmari, I. and Szeles, L.: Decision-making by macrophages and dendritic cells using heterodimeric nuclear receptors to sense their lipid environment, Physiological Reviews 92(2) 739-789, 2012
Nagy, Z., Czimmerer Z. and Nagy, L.: Nuclear receptor mediated mechanisms of macrophage cholesterol metabolism, Cellular and Molecular Endocrinology, 2012
Czimmerer, Z, Varga, T., Poliska, S., Nemet, I., Szanto, A. and Nagy, L.: Identification of novel markers of human alternative macrophage activation including potential endogenous PPARγ ligand production mechanisms, Immunobiology 217: 1301-1314, 2012
Nagy, L: Would eating carrots protect your liver? A new role involving NKT cells for retinoic acid in hepatitis, European Journal of Immunology 42:1677-1680, 2012
Czimmerer Z, Hulvely J, Simandi Z, Varallyay E, Havelda Z, Szabo E, Varga A, Dezso B, Balogh M, Horvath A, Domokos B, Torok Z, Nagy L, Balint BL: A versatile method to design stem-loop primer-based quantitative PCR assays for detecting small regulatory RNA molecules., PLOS ONE 8: (1) e55168, 2013
Dozsa A, Dezso B, Toth BI, Bacsi A, Poliska S, Camera E, Picardo M, Zouboulis CC, Biro T, Schmitz G, Liebisch G, Ruhl R, Remenyik E, Nagy L: PPARgamma-Mediated and Arachidonic Acid-Dependent Signaling is Involved in Differentiation and Lipid Production of Human Sebocytes., J INVEST DERMATOL 1: (1) 1, 2013
Gyongyosi A, Szatmari I, Pap A, Dezso B, Pos Z, Szeles L, Varga T, Nagy L: RDH10, RALDH2 and CRABP2 are required components of PPARg-directed all-trans-retinoic acid synthesis and signaling in human dendritic cells, J LIPID RES 54: (9) 2458-2474, 2013
I Cuaranta-Monroy, Nagy L: PPARy needs a helping hand to make fat, CELL DEATH DIFFER 20: 1599-1600, 2013
Kiss M, Czimmerer Zs, Nagy L: The role of lipid-activated nuclear receptors in shaping macrophage and dendritic cell function - from physiology to pathology, J ALLERGY CLIN IMMUN &: (132) 264-286, 2013
Nagy G, Daniel B, Jonas D, Nagy L, Barta E: A novel method to predict regulatory regions
based on histone mark landscapes
 in macrophages, IMMUNOBIOLOGY 218: (11) 1416-1427, 2013
Nagy Zs, Czimmerer Zs, Nagy L: Pro-inflammatory cytokines negatively regulate PPARg mediated gene expression in both human and murine macrophages via multiple mechanisms, IMMUNOBIOLOGY 218: (11) 1336-1344, 2013
Nagy Zs, Ross J, Rodriguez G, Balint L B, Szeles L, Nagy L, Kirken R A: Genome wide mapping reveals PDE4B as an IL-2 induced STAT5 target, PLOS ONE 8: (2) , 2013
Szenasi T, Kenesi E, Nagy A, Molnar A, Balint BL, Zvara A, Csabai Z, Deak F, Boros Olah B, Mates L, Nagy L, Puskas LG, Kiss I: Hmgb1 can facilitate activation of the matrilin-1 gene promoter by Sox9 and L-Sox5/Sox6 in early steps of chondrogenesis., BBA-GENE REGUL MECH 1829: (10) 1075-1091, 2013
Varga T, Mounier R, Gogolak P, Poliska S, Chazaud B, Nagy L: Tissue LyC6- macrophages are generated in the absence of circulating LyC6- monocytes and Nur77 in a model of muscle regeneration., J IMMUNOL 191: (11) 5695-5701, 2013
Czimmerer Z, Varga T, Poliska S, Nemet I, Szanto A, Nagy L: Identification of novel markers of human alternative macrophage activation including potential endogenous PPARγ ligand production mechanisms, IMMUNOBIOLOGY 217: (12) 1301-1314, 2012
Brazda P, Krieger J, Daniel B, Jonas D, Szekeres T, Langowski J, Tóth K, Nagy L, Vámosi G: Ligand binding shifts highly mobile RXR to chromatin-bound state in a coactivator-dependent manner as revealed by single cell imaging., MOL CELL BIOL 34: (7) 1234-1245, 2014
Cuaranta-Monroy I, Simandi Z, Kolostyak Z, Doan Xuan Quang Minh, Szilard Poliska, Bacso Z, Nagy L: Highly efficient differentiation of embryonic stem cells into adipocytes by ascorbic acid, STEM CELL RES 13: 88-97, 2014
Daniel B, Balint BL, Nagy Zs, Nagy L: Mapping the genomic binding sites of the activated retinoid x receptor in murine bone marrow-derived macrophages using chromatin immunoprecipitation sequencing., METHODS MOL BIOL 1204: 15-24, 2014
Daniel B, Nagy G, Hah H, Horvath A, Czimmerer Z, Poliska P, Gyuris T, Keirsse J, Gysemans C, Ginderachter J, Balint B L, Evans R M, Barta E, Nagy L: The Active Enhancer Network Operated by Liganded RXR Supports Angiogenic Activity in Macrophages, GENE DEV 28: (14) 1562-1577, 2014
Daniel B, Nagy G, Nagy L: The intriguing complexities of mammalian gene regulation: How to link enhancers to regulated genes. Are we there yet?, FEBS LETT 588: (15) 2379-2391, 2014
Dozsa A, Dezso B, Toth BI, Bacsi A, Poliska S, Camera E, Picardo M, Zouboulis CC, Biro T, Schmitz G, Liebisch G, Ruhl R, Remenyik E, Nagy L: PPARgamma-Mediated and Arachidonic Acid-Dependent Signaling is Involved in Differentiation and Lipid Production of Human Sebocytes., J INVEST DERMATOL 134: (4) 910-920, 2014
Daniel B, Nagy G, Hah H, Horvath A, Czimmerer Z, Poliska P, Gyuris T, Keirsse J, Gysemans C, Ginderachter J, Balint B L, Evans R M, Barta E, Nagy L: The Active Enhancer Network Operated by Liganded RXR Supports Angiogenic Activity in Macrophages, GENE DEV 28: (14) 1562-1577, 2014
Gyongyosi A, Docs O, Czimmerer Z, Orosz L, Horvath A, Torok O, Mehes G, Nagy L, Balint BL: Measuring expression levels of small regulatory RNA molecules from body fluids and formalin-fixed, paraffin-embedded samples, METHODS MOL BIOL 1182: 105-119, 2014
Simándi Zoltán, Czipa Erik, Horváth Attila, Kőszeghy Áron, Bordás Csilla, Póliska Szilárd, Juhász István, Imre László, Szabó Gábor, Dezső Balázs, Barta Endre, Sauer Sascha, Károlyi Katalin, Kovács Ilona, Hutóczky Gábor, Bognár László, Klekner Álmos, Szűcs Péter, Bálint Bálint László, Nagy László: PRMT1 and PRMT8 regulate retinoic acid dependent neuronal differentiation with implication to neuropathology, STEM CELLS Epub ahead of print: &, 2014
Daniel B, Nagy G, Nagy L: The intriguing complexities of mammalian gene regulation: How to link enhancers to regulated genes. Are we there yet?, FEBS LETT 588: (15) 2379-2391, 2014
Brignull LM, Czimmerer Z, Saidi H, Daniel B, Villela I, Bartlett NW, Johnston SL, Meira LB, Nagy L, Nohturfft A: Reprogramming of lysosomal gene expression by interleukin-4 and Stat6., BMC GENOMICS 14: , 2013
Daniel B, Nagy G, Hah H, Horvath A, Czimmerer Z, Poliska P, Gyuris T, Keirsse J, Gysemans C, Ginderachter J, Balint B L, Evans R M, Barta E, Nagy L: The Active Enhancer Network Operated by Liganded RXR Supports Angiogenic Activity in Macrophages, GENE DEV 28: (14) 1562-1577, 2014
Gyongyosi A, Docs O, Czimmerer Z, Orosz L, Horvath A, Torok O, Mehes G, Nagy L, Balint BL: Measuring expression levels of small regulatory RNA molecules from body fluids and formalin-fixed, paraffin-embedded samples, METHODS MOL BIOL 1182: 105-119, 2014
Kiss M, Czimmerer Zs, Nagy L: The role of lipid-activated nuclear receptors in shaping macrophage and dendritic cell function - from physiology to pathology, J ALLERGY CLIN IMMUN &: (132) 264-286, 2013
Czimmerer Z, Hulvely J, Simandi Z, Varallyay E, Havelda Z, Szabo E, Varga A, Dezso B, Balogh M, Horvath A, Domokos B, Torok Z, Nagy L, Balint BL: A versatile method to design stem-loop primer-based quantitative PCR assays for detecting small regulatory RNA molecules., PLOS ONE 8: (1) e55168, 2013
Nagy Zs, Ross J, Rodriguez G, Balint L B, Szeles L, Nagy L, Kirken R A: Genome wide mapping reveals PDE4B as an IL-2 induced STAT5 target, PLOS ONE 8: (2) , 2013
Szenasi T, Kenesi E, Nagy A, Molnar A, Balint BL, Zvara A, Csabai Z, Deak F, Boros Olah B, Mates L, Nagy L, Puskas LG, Kiss I: Hmgb1 can facilitate activation of the matrilin-1 gene promoter by Sox9 and L-Sox5/Sox6 in early steps of chondrogenesis., BBA-GENE REGUL MECH 1829: (10) 1075-1091, 2013
Nagy G, Daniel B, Jonas D, Nagy L, Barta E: A novel method to predict regulatory regions
based on histone mark landscapes
 in macrophages, IMMUNOBIOLOGY 218: (11) 1416-1427, 2013
I Cuaranta-Monroy, Nagy L: PPARy needs a helping hand to make fat, CELL DEATH DIFFER 20: 1599-1600, 2013
Nagy, L., Szanto, A., Szatmari, I. and Szeles, L.: 27. Nuclear hormone receptors enable macrophages and dendritic cells to sense their lipid environment and shape their immune response, Physiological Reviews 92(2) 739-789, 2012
Nagy, Z., Czimmerer Z. and Nagy, L.: Nuclear receptor mediated mechanisms of macrophage cholesterol metabolism, Cellular and Molecular Endocrinology, 2012
Nagy, L: Would eating carrots protect your liver? A new role involving NKT cells for retinoic acid in hepatitis, European Journal of Immunology 42:1677-1680, 2012




 

Events of the project

  
2016-07-14 13:02:43
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2015-06-24 08:52:38
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