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Identification of PPARg target genes in macrophages and characterization of their functional roles

English title Identification of PPARg target genes in macrophages and characterization of their functional roles
Applicant Kopf Manfred
Number 182829
Funding scheme Project funding
Research institution Molekulare Gesundheitswissenschaften Departement Biologie ETH Zürich
Institution of higher education ETH Zurich - ETHZ
Main discipline Molecular Biology
Start/End 01.10.2018 - 31.12.2021
Approved amount 912'035.00
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All Disciplines (2)

Discipline
Molecular Biology
Immunology, Immunopathology

Keywords (4)

Alveolar Macrophages; Red pulp macrophages; Peroxisome Proliferator Activated Receptor (PPARg); fetal monocyte precursor

Lay Summary (German)

Lead
Identifikation und Charakterisierung der Funktion Genen, die von dem Transkriptionsfaktor PPARg in der Entwicklung von Gewebemakrophagen reguliert werden.
Lay summary

In dem Organ und Gewebe des Körpers gibt es Subpopulationen von Makrophagen, sogenannte residente Gewebemakrophagen (GeMac). Neben einer immunologischen Funktion in der Abwehr von Mikroorganismen, die alle GeMac gemeinsam besitzen, gibt es organspezifische Funktionen der jeweiligen GeMac zur Erhaltung der Homöostase. Beispielsweise sind alveolare Makrophagen (alvMac) in der Lunge verantwortlich für den Stoffwechsel von Lipoproteinen und Phospholipiden (surfacant) in den Alveolen, um deren Integrität zu bewahren und somit den Gasaustausch einwandfrei zu gewährleisten. Erst vor wenigen Jahren hat man herausgefunden, dass GeMac schon sehr früh in der Embryogenese entstehen. Der wirkliche Herkunftsort im Embryo belibt jedoch umstritten, wobei primitive Macrophagen des Dottersacks oder fötale Monocyten der Leber als Vorläuferzellen vorgeschlagen wurden. Um diese Frage zu beantworten haben wir die Vorläuferzellen aus dem Dottersack und der fötalen Leber isoliert und in Mäuse transferiert, die aufgrund eines genetischen Defektes (Csf2R-/-) keine eigenen alveolare Makrophagen haben. Wir hatten in der Vergangenheit gezeigt, dass alveolare Makrophagen in Csf2R-/- Mäuse entstehen können, wenn man ihnen nach der Geburt fötale Monocyten aus der Lunge von Wild-typ Mäusen spendet (transferiert). Wir konnten dieses Ergebnis bestätigen mit dem Transfer von Monocyten die aus der fötalen Leber präpariert wurden. Im Gegensatz dazu konnten primitive Makrophagen aus dem Dottersack die Entwicklung alvMac in Csf2R-/- Mäuse viel weniger gut rekonstituieren. Auch war deren Funktion eingeschränkt. Dieses Ergebnis deutet darauf hin, dass alveolare Makrophagen und viele andere Gewebemakrophagen aus Monocyten Stammzellen der fötalen Leber entstehen.
Desweiteren haben wir vor einigen Jahren gezeigt, dass der Faktor (Csf2) welcher von Epithelzellen der Alveoli produziert wird, in fötalen Monocyten der Lunge den Transkriptionsfaktor PPARg aktiviert und dieser essentiell ist für die Entwicklung von alvMac. Auf die Frage, ob andere Arten von Gewebemarkrophagen eventuell auch PPARg brauchen, fanden wir eine Population von Makrophagen die in der roten Pulpa der Milz lokalisiert sind, welche tatsächlich ohne PPARg nicht entsehen können. Momentan untersuchen wir die molekularen Mechanismen.

Direct link to Lay Summary Last update: 26.10.2018

Lay Summary (English)

Lead
Identification of PPARg target genes in macrophages and characterization of theirfunctional roles
Lay summary

Almost every tissue and organ of the body contains a subset of tissue-resident macrophages. They are responsible for clearance of invading microorganisms and cellular debris, thereby maintaining tissue homeostasis. Moreover, they are involved in tissue repair and exert tissue specific functions such as catabolism of lung surfactant by alveolar macrophages (AM). It is now well-accepted that the majority of tissue macrophage subsets originate in the embryo and are thereafter maintained by local self-renewal. We reported that GM-CSF produced by lung epithelial cells instructs AM development by induction of the transcription factor PPARg in fetal lung monocyte around the time of birth in mice. In contrast, resident macrophages in almost every other tissue develop in a PPARg-independent manner. We found that development of both splenic red pulp macrophages (RPM) and bone erythroid island macrophages (BMEIM) also require PPARg. Otherwise, RPM and BMEIM require the transcription factor SpiC and its repressor BACH1. Surprisingly, we found that PPARg and SpiC/BACH1 appear to be separate pathways that do not interact directly. We are currently studying the genes regulated by PPARg in AM, RPM, and BMEIM by applying ChipSeq and by comparing the transcriptome of WT and Pparg-deficient AM precursors. We identified a few candidate genes, which will be validate by generation of conditional knockout mice. Moreover, we have established a method allowing long-term in vitro culture of fetal liver monocytes (fliMo), which can differentiate to AM upon intranasal installation into newborns. We suggested to use Crispr/Cas9 for high-throughput gene editing in fliMo and transfer manipulated cells to identify genes required for development and function of AM. Unfortunately, this method turned out to be very inefficient because of low transfection efficiency of fliMo. We are currently working on an alternative approach using bone marrow stem cells instead of fliMo for Cripr/Cas9 gene editing.
In a related project, we have shown that the large majority of AM are ablated during influenza virus infection and are replaced by invading bone marrow derived blood monocytes, which can differentiate into AM.  We are addressing the question whether fetal liver derived AM or blood monocyte derived AM are qualitatively different in their function in respiratory infections.




Direct link to Lay Summary Last update: 26.10.2018

Responsible applicant and co-applicants

Employees

Publications

Publication
Loss of Rnf31 and Vps4b sensitizes pancreatic cancer to T cell-mediated killing
Frey Nina, Tortola Luigi, Egli David, Janjuha Sharan, Rothgangl Tanja, Marquart Kim Fabiano, Ampenberger Franziska, Kopf Manfred, Schwank Gerald (2022), Loss of Rnf31 and Vps4b sensitizes pancreatic cancer to T cell-mediated killing, in Nature Communications, 13(1), 1804-1804.
Peroxisomes Are Critical for the Development and Maintenance of B1 and Marginal Zone B Cells but Dispensable for Follicular B Cells and T Cells
Muri Jonathan, Corak Basak, Matsushita Mai, Baes Myriam, Kopf Manfred (2022), Peroxisomes Are Critical for the Development and Maintenance of B1 and Marginal Zone B Cells but Dispensable for Follicular B Cells and T Cells, in The Journal of Immunology, 208(4), 839-850.
PPARɣ drives IL-33-dependent ILC2 pro-tumoral functions
Ercolano Giuseppe, Gomez-Cadena Alejandra, Dumauthioz Nina, Vanoni Giulia, Kreutzfeldt Mario, Wyss Tania, Michalik Liliane, Loyon Romain, Ianaro Angela, Ho Ping-Chih, Borg Christophe, Kopf Manfred, Merkler Doron, Krebs Philippe, Romero Pedro, Trabanelli Sara, Jandus Camilla (2021), PPARɣ drives IL-33-dependent ILC2 pro-tumoral functions, in Nature Communications, 12(1), 2538-2538.
Differential sensitivity of inflammatory macrophages and alternatively activated macrophages to ferroptosis
Piattini Federica, Matsushita Mai, Muri Jonathan, Bretscher Peter, Feng Xiaogang, Freigang Stefan, Dalli Jesmond, Schneider Christoph, Kopf Manfred (2021), Differential sensitivity of inflammatory macrophages and alternatively activated macrophages to ferroptosis, in European Journal of Immunology, 51(10), 2417-2429.
Comparative analysis of the role of mast cells in murine asthma models using Kit‐sufficient mast cell‐deficient animals
Pohlmeier Lea, Sonar Sanchaita Sriwal, Rodewald Hans‐Reimer, Kopf Manfred, Tortola Luigi (2021), Comparative analysis of the role of mast cells in murine asthma models using Kit‐sufficient mast cell‐deficient animals, in Allergy, 76(7), 2030-2043.
Redox regulation of immunometabolism
Muri Jonathan, Kopf Manfred (2021), Redox regulation of immunometabolism, in Nature Reviews Immunology, 21(6), 363-381.
GM-CSF instigates a dendritic cell–T-cell inflammatory circuit that drives chronic asthma development
Nobs Samuel Philip, Pohlmeier Lea, Li Fengqi, Kayhan Merve, Becher Burkhard, Kopf Manfred (2021), GM-CSF instigates a dendritic cell–T-cell inflammatory circuit that drives chronic asthma development, in Journal of Allergy and Clinical Immunology, 147(6), 2118-2133.e3.
Tissue-resident macrophages: guardians of organ homeostasis
Nobs Samuel Philip, Kopf Manfred (2021), Tissue-resident macrophages: guardians of organ homeostasis, in Trends in Immunology, 42(6), 495-507.
PPARγ is essential for the development of bone marrow erythroblastic island macrophages and splenic red pulp macrophages
Okreglicka Katarzyna, Iten Irina, Pohlmeier Lea, Onder Lucas, Feng Qian, Kurrer Michael, Ludewig Burkhard, Nielsen Peter, Schneider Christoph, Kopf Manfred (2021), PPARγ is essential for the development of bone marrow erythroblastic island macrophages and splenic red pulp macrophages, in Journal of Experimental Medicine, 218(5), 1-16.
Comprehensive characterization of myeloid cells during wound healing in healthy and healing‐impaired diabetic mice
Joshi Natasha, Pohlmeier Lea, Ben‐Yehuda Greenwald Maya, Haertel Eric, Hiebert Paul, Kopf Manfred, Werner Sabine (2020), Comprehensive characterization of myeloid cells during wound healing in healthy and healing‐impaired diabetic mice, in European Journal of Immunology, 50(9), 1335-1349.
High-Dimensional T Helper Cell Profiling Reveals a Broad Diversity of Stably Committed Effector States and Uncovers Interlineage Relationships
Tortola Luigi, Jacobs Andrea, Pohlmeier Lea, Obermair Franz-Josef, Ampenberger Franziska, Bodenmiller Bernd, Kopf Manfred (2020), High-Dimensional T Helper Cell Profiling Reveals a Broad Diversity of Stably Committed Effector States and Uncovers Interlineage Relationships, in Immunity, 53(3), 597-613.e6.
Endothelial Lactate Controls Muscle Regeneration from Ischemia by Inducing M2-like Macrophage Polarization
Zhang Jing, Muri Jonathan, Fitzgerald Gillian, Gorski Tatiane, Gianni-Barrera Roberto, Masschelein Evi, D’Hulst Gommaar, Gilardoni Paola, Turiel Guillermo, Fan Zheng, Wang TongTong, Planque Mélanie, Carmeliet Peter, Pellerin Luc, Wolfrum Christian, Fendt Sarah-Maria, Banfi Andrea, Stockmann Christian, Soro-Arnáiz Inés, Kopf Manfred, De Bock Katrien (2020), Endothelial Lactate Controls Muscle Regeneration from Ischemia by Inducing M2-like Macrophage Polarization, in Cell Metabolism, 31(6), 1136-1153.e7.
Glutathione peroxidase 4 and vitamin E control reticulocyte maturation, stress erythropoiesis and iron homeostasis
Altamura Sandro, Vegi Naidu M., Hoppe Philipp S., Schroeder Timm, Aichler Michaela, Walch Axel, Okreglicka Katarzyna, Hültner Lothar, Schneider Manuela, Ladinig Camilla, Kuklik-Roos Cornelia, Mysliwietz Josef, Janik Dirk, Neff Frauke, Rathkolb Birgit, de Angelis Mar tin Hrabé, Buske Christian, Silva Ana Rita da, Muedder Katja, Conrad Marcus, Ganz Tomas, Kopf Manfred, Muckenthaler Martina U., Bornkamm Georg W. (2020), Glutathione peroxidase 4 and vitamin E control reticulocyte maturation, stress erythropoiesis and iron homeostasis, in Haematologica, 105(4), 937-950.
Cyclopentenone Prostaglandins and Structurally Related Oxidized Lipid Species Instigate and Share Distinct Pro- and Anti-inflammatory Pathways
Muri Jonathan, Feng Qian, Wolleb Helene, Shamshiev Abdijapar, Ebner Christian, Tortola Luigi, Broz Petr, Carreira Erick M., Kopf Manfred (2020), Cyclopentenone Prostaglandins and Structurally Related Oxidized Lipid Species Instigate and Share Distinct Pro- and Anti-inflammatory Pathways, in Cell Reports, 30(13), 4399-4417.e7.
Fetal monocytes possess increased metabolic capacity and replace primitive macrophages in tissue macrophage development
Li Fengqi, Okreglicka Katarzyna Maria, Pohlmeier Lea Maria, Schneider Christoph, Kopf Manfred (2020), Fetal monocytes possess increased metabolic capacity and replace primitive macrophages in tissue macrophage development, in The EMBO Journal, 39(3), 1-16.
B1 and Marginal Zone B Cells but Not Follicular B2 Cells Require Gpx4 to Prevent Lipid Peroxidation and Ferroptosis
Muri Jonathan, Thut Helen, Bornkamm Georg W., Kopf Manfred (2019), B1 and Marginal Zone B Cells but Not Follicular B2 Cells Require Gpx4 to Prevent Lipid Peroxidation and Ferroptosis, in Cell Reports, 29(9), 2731-2744.e4.
Deciphering CD4+ T cell specificity using novel MHC–TCR chimeric receptors
Kisielow Jan, Obermair Franz-Josef, Kopf Manfred (2019), Deciphering CD4+ T cell specificity using novel MHC–TCR chimeric receptors, in Nature Immunology, 20(5), 652-662.
GM-CSF intrinsically controls eosinophil accumulation in the setting of allergic airway inflammation
Nobs Samuel Philip, Kayhan Merve, Kopf Manfred (2019), GM-CSF intrinsically controls eosinophil accumulation in the setting of allergic airway inflammation, in Journal of Allergy and Clinical Immunology, 143(4), 1513-1524.e2.
A Macrophage-Pericyte Axis Directs Tissue Restoration via Amphiregulin-Induced Transforming Growth Factor Beta Activation
Minutti Carlos M., Modak Rucha V., Macdonald Felicity, Li Fengqi, Smyth Danielle J., Dorward David A., Blair Natalie, Husovsky Connor, Muir Andrew, Giampazolias Evangelos, Dobie Ross, Maizels Rick M., Kendall Timothy J., Griggs David W., Kopf Manfred, Henderson Neil C., Zaiss Dietmar M. (2019), A Macrophage-Pericyte Axis Directs Tissue Restoration via Amphiregulin-Induced Transforming Growth Factor Beta Activation, in Immunity, 50(3), 645-654.e6.

Collaboration

Group / person Country
Types of collaboration
Philipp Henneke, Med Faculty, University Freiburg Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Exchange of personnel
Ueli Suter, Dept Biology, ETH Zürich Switzerland (Europe)
- Publication
Wolf-Dietrich Hardt, Dept Biology, ETH Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Camilla Jandus, Med Faculty, University Geneva Switzerland (Europe)
- Publication
Gerald Schwank, Med Faculty, University Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Dietmar Zaiss, University Regensburg Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Exchange of personnel
Katrin De Bock, Dept HEST, ETH Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Sabine Werner, Dept Biology, ETH Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Timo Speer, Med Faculty, University Saarland Germany (Europe)
- Publication

Communication with the public

Communication Title Media Place Year
Media relations: print media, online media Braucht es die Corona-Impfung für Kinder Blick German-speaking Switzerland 2021
Media relations: print media, online media Bundesrat muss Rückkehr zur Normalität wohl verschieben 20 Minuten German-speaking Switzerland 2021
Media relations: print media, online media Corona könnte in der Schweiz im Frühling vorbei sein 20 Minuten German-speaking Switzerland 2021
Media relations: print media, online media Coronavirus: Ist die Herdenimmunität noch möglich? NAU -News für die Schweiz German-speaking Switzerland 2021
Media relations: print media, online media Der Druck auf Ungeimpfte wird zunehmen 20 Minuten German-speaking Switzerland 2021
Media relations: radio, television Die erste präventive Corona-Therapie SRF German-speaking Switzerland 2021
Media relations: print media, online media Dritte Impfung als Chance für einen Teil der Immungeschwächten SRF German-speaking Switzerland 2021
Media relations: radio, television Eine dritte Corona-Impfung für Menschen mit Immunschwäche? SRF German-speaking Switzerland 2021
Talks/events/exhibitions Erfolge in der Behandlung von Volkskrankheiten durch Forschung anTieren German-speaking Switzerland 2021
Media relations: print media, online media Geimpfte und Genesene werden auch bei Omikron einen Vorteil haben» 20 Minuten German-speaking Switzerland 2021
Media relations: print media, online media Geopolitisch», «ideologisch» – schwere Kritik am BAG wegen Impf-Politik 20 Minuten German-speaking Switzerland 2021
Media relations: print media, online media Hoffnung auf Herdenimmunität dank Omikron ist ein Wunschtraum» 20Minuten German-speaking Switzerland 2021
Media relations: print media, online media https://www.20min.ch/story/die-corona-zahlen-koennten-in-drei-wochen-alle-rekorde-brechen-1750361795 20 Minuten German-speaking Switzerland 2021
Media relations: print media, online media Ich vertraue europäischen Impfungen mehr als der russischen 20 Minuten German-speaking Switzerland 2021
Media relations: print media, online media Masken, Booster, 3G im Job – so wollen Experten die 5. Welle stoppen 20 Minuten German-speaking Switzerland 2021
Media relations: print media, online media Ohne Impfung hätten wir in Kürze Tausende Corona-Tote mehr 20 Minuten German-speaking Switzerland 2021
Media relations: print media, online media So will das BAG Ungeimpfte noch überzeugen 20 Minuten German-speaking Switzerland 2021
Media relations: print media, online media Solltest du dich jetzt impfen lassen oder auf Omikron-Impfstoff warten? 20 Minuten German-speaking Switzerland 2021
Media relations: print media, online media Astra-Zeneca nimmt Impfstoffstudie wieder auf – so geht es jetzt weiter NZZ German-speaking Switzerland 2020
Media relations: print media, online media Ein Drittel der Personen hat Gedächtnis-Immunzellen, die das neue Virus erkennen NZZ German-speaking Switzerland 2020
Media relations: print media, online media Im April ist dank Impfstoff ein halbwegs normales Leben möglich 20 Minuten German-speaking Switzerland 2020
Media relations: print media, online media Kommt die kleine Schweiz beim Corona-Impfstoff zu kurz? 20 Minuten German-speaking Switzerland 2020
Talks/events/exhibitions Tierversuche retten Menschenleben German-speaking Switzerland 2019

Associated projects

Number Title Start Funding scheme
163443 Development of alveolar macrophages and splenic red pulp macrophages 01.10.2015 Project funding

Abstract

PPARa, PPARb/d, and PPARg are members of the Peroxisome Proliferator-Activated Receptor (PPAR) subfamily of nuclear receptors and transcription factors. For activity, the require binding of a lipid ligand and dimerization with a retinoid acid receptor (RXR) subfamily member for high-affinity DNA binding to PPAR response elements of target gene promoters. PPARs play central roles in lipid metabolism, fatty acid oxidation, adipogenesis, and energy metabolism. Both PPARb/d and PPARg also have anti-inflammatory activities. We have previously reported an essential role of GM-CSF and PPARg in development of alveolar macrophages (AM) and a requirement of PPARg in both DCs and CD4+ T cells for development allergic airway type 2 responses. Moreover, we recently unveiled a PPAR necessity for development of red pulp macrophages. Proposed future studies:-Having established that fetal monocytes are superior to primitive macrophages in their capacity to develop into AM, we are currently studying the underlying molecular mechanism. Preliminary data indicate differences in metabolism, which we want to strengthen by measurement of various metabolic parameters and metabolites. -RNA sequencing of fetal lung monocyte precursors identified a few genes that are downregulated in absence of both GM-CSF and PPARg suggesting that they are potentially involved in GM-CSF/PPARg mediated AM development. To study the definitive role of these genes, we will perform loss-of-function studies.- We are establishing a novel cell culture model for high-thoughput gene manipulation of primary monocytes/macrophages and their consequences for AM development and function. - To verify whether genes downregulated in the absence of PPARg in fetal lung monocyte precursors, we are establishing a novel genome-scale chromatin profiling strategy for high-resolution mapping of DNA binding sites in low number of primary cells such as alveolar macrophages and dendritic cells. - Finally, we are generating mice lacking PPARb/d specifically in T cells and myeloid cells to define its intrinsic roles. Taken together, the results of these studies will help to unveil the functions of PPARs in the immune system and thereby link fundamental cellular lipid metabolism and immunity.
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