Extracellular and intracellular adenine nucleotides (AN) impact on all central processes in biology and medicine. AN are essential and ubiquitous signaling molecules involved in regulating universal cellular processes, including (i) cell-cell communication and (ii) intracellular signaling.
Unresolved issues regarding the signaling function of extracellular AN in inflammation, e.g. adenosine triphosphate (ATP) or nicotinamide adenine dinucleotide (NAD), relate to the timing and location of their release, their conversion by ecto-enzymes, and their biological role within the balance of inflammatory processes. Likewise, the precise role of intracellular AN second messengers, e.g. nicotinic acid adenine dinucleotide phosphate (NAADP) or 3’,5’-cyclic adenosine monophosphate (cAMP), in the spatio-temporal control of signaling processes by forming or modulating microdomains with their metabolizing enzymes, specific binding proteins or receptors, or target ion channels remains largely unknown.
The central goal of the research consortium is to further our understanding of the regulatory roles of AN and their kinetics in the context of inflammatory diseases. Specific aims relate to (i) modulation of the balance between pro- and anti-inflammatory processes by AN converting ecto-nucleotidases and purinergic receptors, and to (ii) AN-driven intracellular calcium signaling and cAMP signaling in inflammation.
POSITIONS OFFERED FOR SECOND FUNDING PERIOD
Please apply directly on the career pages of the universities
Post Doc & PhD Student
A01 – Andreas H. Guse (UKE)
In the Department of Biochemistry and Molecular Cell Biology at the University Medical Center Hamburg-Eppendorf, one of the subprojects of A01 (with Alexander Flügel, University of Göttingen) works on the importance of adenine nucleotides for the activation of T cells. This research has been ongoing for several years (see Wolf et al., Science Signal. 2015, Diercks et al, Science Signal. 2018, Roggenkamp et al, 2021, Gu et al, Science Signal. 2021).
First experience cell culture, high-resolution fluorescence microscopy, HPLC analysis, molecular cloning techniques, protein biochemistry would be advantageous
A03 – Hans-Willi Mittrücker (UKE)
One focus of the work at the Institute of Immunology (Hans-Willi Mittrücker) and the I. Medical Clinic and Polyclinic (Samuel Huber) of the University Medical Center Hamburg-Eppendorf is the regulation and function of T lymphocytes in inflammation and infection (Brockmann et al. Nat Commun 2018, Perez et al. Nat Commun 2020, Krebs et al. Sci Immunol 2020).
First experience in cell culture, flow cytometry, infection models would be advantageous
A05 – Ralf Fliegert (UKE)
In the Department of Biochemistry and Molecular Cell Biology at the University Medical Center Hamburg-Eppendorf, one of the subprojects of A05 (with Maria Garcia Alai, EMBL) investigates the characterization of the adenine nucleotide-gated ion channel TRPM2 structurally and functionally. This work has been ongoing for several years (see Fliegert et al., Nat Chem Biol. 2017, Gattkowski et al, BBA Mol Cell Res. 2019, Riekehr et al, IJMS 2022, Sander et al, Protein Sci. 2022).
First experience cell culture, fluorescence microscopy, HPLC analysis, molecular cloning techniques, protein biochemistry would be advantageous
A07 – Christine Gee (UKE)
The collaborative project A07, headed by Dr. Christine Gee (ISP) and Prof. Christian Lohr (Uni. Hamburg) focuses on the investigation of astrocyte-neuron interactions and their control by adenine nucleotides in connection with the pathogenesis of Multiple Sclerosis and its animal model.
The position is about the role of cAMP in astrocyte-neuron interactions using state of the art optogenetic tools, electrophysiology and 2-photon laser scanning microscopy. The project focuses on understanding information processing and storage by individual synapses and the development of new optogenetic tools (see Yang et al., 2021 BMC Biol 19(1):227; Anisimova et al., 2022 Cereb Cortex doi: 10.1093/cercor/bhac050; Perez-Alvarez et al., 2020 Nat Commun 11(1):2464) .
A07 – Christian Lohr (UHH)
The Division of Neurophysiology investigates the role of astrocytes in brain function and pathophysiology (neuroinflammation). We employ two-photon/confocal calcium and cAMP imaging combined with patch-clamp electrophysiology, optogenetics and immunohistology. The successful candidate will be part of the team that uses these techniques to study second messenger signaling in astrocytes in acute brain slices of the hippocampus and the olfactory bulb. Performing and analysing imaging experiments as well as antibody staining of fixed tissue will be part of the duties.
A12 – Julian Schulze zur Wiesch
A13 – Jakob Körbelin (UKE)
As part of a subproject with Tim Magnus (UKE) of the SFB 1328, the Department of Neurology has been investigating the importance of adenine nucleotides in stroke for several years. Starting with the current funding period, the ENDomics (Oncology) group will participate in this subproject with the development of targeted AAV vectors for the modulation of adenine nucleotide-converting enzymes in the mouse model of stroke.
A15 – Annette Nicke (LMU)
The “Ion channel receptor lab” in the Walther-Straub-Institute of Pharmacology and Toxicology at the Ludwig-Maximilians-Universität München is an interdisciplinary and international group and integrated in several research networks.
To identify the molecular mechanisms of P2X7 signaling (Kopp et al. Front. Mol. Neurosci.), we use a novel P2X7-EGFP BAC transgenic mouse model (Kaczmarek-Hájek, Zhang, Kopp et al., eLife, 2018) in combination with biochemical (cross-linking mass spectrometry) and biophysical (two-electrode voltage clamp, voltage clamp fluorometry) approaches. We are looking for a highly motivated person with enthusiasm for basic science and a good background in molecular biology and biochemistry. Experience with genetically modified mice and/or TEVC, VCF, or Ca2+ imaging is an advantage.
A20 - Pablo Sáez (UKE)
The Cell Communication and Migration Laboratory (CellComM Lab) is hosted in the Department of Biochemistry and Molecular Cell Biology (IBMZ). We combine live-cell imaging with microfluidics and advance imaging analysis to study how cell move and communicate during health and disease (Sáez et al 2017 Sci Signal; Bretou, Sáez et al 2017 Sci Immunol; https://youtu.be/pOO4XmKlTls?t=1752). This project aims to unveil the complex regulation of adenine nucleotides over leukocyte migration both in vitro and in vivo, in the context of adipose tissue inflammation.
Postdoc & PhD Student
A21 - Marc Freichel University of Heidelberg
In the Department of Pharmacology at the University of Heidelberg (new partproject in 2nd funding period with Christian Grimm, LMU), we study Ca2+ dependent activation of mast cells. To this end we combine generation of transgenic mouse lines using genome editing with Ca2+ imaging and electrophysiological approaches to study Ca2+ homeostasis. Ca2+-dependent transcriptional gene programmes triggered by adenine nucleotides are analyze with transcription analyses.
First experience cell culture, fluorescence microscopy, molecular cloning techniques would be advantageous, and willingness to work with laboratory animals is necessary.
PAPER SCIENCE ADVANCES
ATP is not only an essential energy-source but also an important singling molecule during inflammation. In this Publication Brock et al. show that initial Ca2+ microdomains are significantly decreased in T cells where ATP-gated cation channels are knock-out, namely P2X4 and P2X7. This effect was already quantifiable during the first second after TCR stimulation and could be verified by pharmacological inhibition of P2X4 or P2X7 blocking nanobodies as well as extracellular Apyrase and blocking of Pannexin-1. This work was part of the project A02 (Diercks/Werner) in close collaboration with projects A01, A03, A11 and Z02.
PAPER SCIENCE SIGNALING
Early steps in T cell activation are mediated by the synthesis of Ca2+-mobilizing second messenger NAADP. In this publication NAADPH-oxidizing enzymes that were critical for the early phases of T cell activation were identified. In cultured rat T cells, knockout of DUOX2 reduced local Ca2+ microdomain formation, whereas functional knockout of both DUOX1 and DUOX2 in murine T cells suppressed global intracellular Ca2+ signaling. This work was part of project A01 and A02 in close collaboration with projects A03, A04 and A10.
For further announcements see:
Hosang L, Canals RC, van der Flier FJ, Hollensteiner J, Daniel R, Flügel A, Odoardi F (2022) The lung microbiome regulates brain autoimmunity. Nature. 603(7899):138-144.
Brock VJ, Wolf IMA, Er-Lukowiak M, Lory N, Stähler T, Woelk LM, Mittrücker HW, Müller CE, Koch-Nolte F, Rissiek B, Werner R, Guse AH, Diercks BP. P2X4 and P2X7 are essential players in basal T cell activity and Ca2+ signaling milliseconds after T cell activation. Sci Adv. 2022 Feb 4;8(5):eabl9770.
Gil D, Diercks BP, Guse AH, Dupont G (2022) Three-Dimensional Model of Sub-Plasmalemmal Ca2+ Microdomains Evoked by T Cell Receptor/CD3 Complex Stimulation. Front Mol Biosci. 9:811145.
Zickler M, Stanelle-Bertram S, Ehret S, Heinrich F, Lange P, Schaumburg B, Kouassi NM, Beck S, Jaeckstein MY, Mann O, Krasemann S, Schroeder M, Jarczak D, Nierhaus A, Kluge S, Peschka M, Schlüter H, Renné T, Pueschel K, Kloetgen A, Scheja L, Ondruschka B, Heeren J, Gabriel G (2022) Replication of SARS-CoV-2 in adipose tissue determines organ and systemic lipid metabolism in hamsters and humans. Cell Metab. 34:1-2.
Di Lauro C, Bianchi C, Sebastián-Serrano Á, Soria-Tobar L, Alvarez-Castelao B, Nicke A, Díaz-Hernández M (2022) P2X7 receptor blockade reduces tau induced toxicity, therapeutic implications in tauopathies. Prog Neurobiol. 208:102173
Schneider E, Winzer R, Rissiek A, Ricklefs I, Meyer-Schwesinger C, Ricklefs FL, Bauche A, Behrends J, Reimer R, Brenna S, Wasielewski H, Lauten M, Rissiek B, Puig B, Cortesi F, Magnus T, Fliegert R, Müller CE, Gagliani N, Tolosa E (2021) CD73-mediated adenosine production by CD8 T cell-derived extracellular vesicles constitutes an intrinsic mechanism of immune suppression. Nat Commun 12, 5911.
Gu F, Krüger A, Roggenkamp HG, Alpers R, Lodygin D, Jaquet V, Möckl F, Hernandez C. LC, Winterberg k, Bauche A, Rosche A, Grasberger H, Kao JY, Schetelig D, Werner R, Schröder K, Carty M, Bowie AG, Huber S, Meier C, Mittrücker HW, Heeren J, Krause KH, Flügel A, Diercks BP, Guse AH (2021) Dual NADPH oxidases DUOX1 and DUOX2 synthesize NAADP and are necessary for Ca2+ signaling during T cell activation. Sci. Signal. 14, eabe3800.
Kurelic R, Krieg PF, Sonner JK, Bhaiyan G, Ramos GC, Frantz S, Friese MA, Nikolaev VO (2021) Upregulation of Phosphodiesterase 2A Augments T Cell Activation by Changing cGMP/cAMP Cross-Talk. Front. Pharmacol. 12:748798.
Yang S*, Constantin OM*, Sachidanandan D*, Hofmann H, Kunz TC, Kozjak-Pavlovic V, Oertner TG, Nagel G, Kittel RJ, Gee CE*, Gao S* (2021) PACmn for improved optogenetic control of intracellular cAMP. BMC Biol. 19:227.
Baum N, Eggers M, Koenigsdorf J, Menzel S, Hambach J, Staehler T, Fliegert R, Kulow F, Adam G, Haag F, Bannas P, Koch-Nolte F (2021) Mouse CD38-Specific Heavy Chain Antibodies Inhibit CD38 GDPR-Cyclase Activity and Mediate Cytotoxicity Against Tumor Cells. Front Immunol. 12:703574.
Müller CE, Namasivayam V. Recommended tool compounds and drugs for blocking P2X and P2Y receptors. Purinergic Signal. 2021 online ahead of print