A01

A01: Summary

From Diercks, B.-P. et al. (2018). ORAI1, STIM1/2, and RYR1 shape subsecond Ca2+ microdomains upon T cell activation. Sci. Signal. 11, eaat0358.

In the previous project we obtained evidence for major roles of ryanodine receptor (RyR) 1 and the 2nd messenger NAADP in T cell biology and RyR3 in astrocyte biology. Using RyR1 and RyR3 knock-out mice we found ameliorated EAE course compared to control animals. Further, we designed and applied a novel biosensor for NFAT translocation and used high resolution Ca2+ imaging to characterize local initial Ca2+ signals. Here we aim to pursue the biochemical basis of NAADP action by identifying NAADP binding proteins and the NAADP forming enzyme. Further we will analyze in detail the role of NAADP, NAADP binding proteins and RyR1 in initial local Ca2+ signals during immune synapse initiation. T cells from a conditional knock-out model for RyR 1 will be used to study local Ca2+ signals, active experimental autoimmune encephalomyelitis (EAE) and transfer EAE, and T cell activation in the lymph node and central nervous system (CNS) by 2-photon imaging. Further, mice with RyR1 deficiency in astrocytes (GFAP-Cre), neurons (Thy1-Cre) and microglia (Cx3cr1-CreER) will be generated and their phenotyping under steady state and inflammatory conditions will be studied. In addition, we will analyze global and local Ca2+ signaling in wt vs RyR3-/- murine astrocytes and perform gene expression profiling (RNA-Seq) of Ryr3-deficient astrocytes isolated ex vivo from healthy or inflamed CNS. We expect to precisely describe the signaling axis involving NAADP and RyR1 in T cells and its role in T cell activation and CNS autoimmunity. Likewise, we expect to unravel the role of RyR3 signaling in astrocytes and their role in CNS autoimmunity.

Prof. Dr. Dr. Andreas H. Guse

Selected publications

 

  1. Fliegert R, Bauche A, Wolf Pérez AM, Watt JM, Rozewitz MD, Winzer R, Janus M, Gu F, Rosche A, Harneit A, Flato M, Moreau C, Kirchberger T, Wolters V, Potter BVL, Guse AH (2017) 2'Deoxyadenosine 5' diphosphoribose is an endogenous TRPM2 superagonist. Nat Chem Biol 13:10361044.
  2. Wolf IM, Diercks BP, Gattkowski E, Czarniak F, Kempski J, Werner R, Schetelig D, Mittrücker HW, Schumacher V, von Osten M, Lodygin D, Flügel A, Fliegert R, Guse AH (2015) Frontrunners of T cell activation: Initial, localized Ca2+ signals mediated by NAADP and the type 1 ryanodine receptor. Sci Signal 8:ra102.
  3. Nebel M, Schwoerer AP, Warszta D, Siebrands CC, Limbrock AC, Swarbrick JM, Fliegert R, Weber K, Bruhn S, Hohenegger M, Geisler A, Herich L, Schlegel S, Carrier L, Eschenhagen T, Potter BV, Ehmke H, Guse AH (2013) Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP)-mediated Calcium Signaling and Arrhythmias in the Heart Evoked by β-Adrenergic Stimulation. J Biol Chem 288:1601716030.
  4. Schmid F, Fliegert R, Westphal T, Bauche A, Guse AH (2012) Nicotinic acid adenine dinucleotide phosphate (NAADP) degradation by alkaline phosphatase. J Biol Chem 287:32525-32534.
  5. Cordiglieri C, Odoardi F, Zhang B, Nebel M, Kawakami N, Klinkert WE, Lodygin D, Lühder F, Breunig E, Schild D, Ulaganathan VK, Dornmair K, Dammermann W, Potter BV*, Guse AH*, Flügel A* (2010) Nicotinic acid adenine dinucleotide phosphate-mediated calcium signalling in effector T cells regulates autoimmunity of the central nervous system. Brain 133:1930-1943. *equal contribution as last authors
  6. Dammermann W, Zhang B, Nebel M, Cordiglieri C, Odoardi F, Kirchberger T, Kawakami N, Dowden J, Schmid F, Dornmair K, Hohenegger M, Flügel* A, Guse AH*, Potter BV* (2009) NAADP-mediated Ca2+ signaling via type 1 ryanodine receptor in T cells revealed by a synthetic NAADP antagonist. Proc Natl Acad Sci USA 106:10678-10683. *equal contribution as last authors
  7. Gasser A, Bruhn S, Guse AH (2006) Second messenger function of nicotinic acid adenine dinucleotide phosphate revealed by an improved enzymatic cycling assay. J Biol Chem 281:1690616913.
  8. Bruzzone S, Kunerth S, Zocchi E, De Flora A, Guse AH (2003) Spatio-temporal propagation of Ca2+ signals by cyclic ADP-ribose in 3T3 cells stimulated via purinergic P2Y receptors. J Cell Biol 163:837845.
  9. Berg I, Potter BV, Mayr GW, Guse AH. (2000) Nicotinic acid adenine dinucleotide phosphate (NAADP+) is an essential regulator of T-lymphocyte Ca2+-signaling. J Cell Biol 150:581-588.
  10. Guse AH, da Silva CP, Berg I, Skapenko AL, Weber K, Heyer P, Hohenegger M, Ashamu GA, Schulze-Koops H, Potter BV, Mayr GW (1999) Regulation of calcium signalling in T lymphocytes by the second messenger cyclic ADP-ribose. Nature 398:70-73.

 

 

 

Prof. Dr. Alexander Flügel

Selected publications

 

  1. Lodygin D*, Hermann M*, Schweingruber N, Flügel-Koch C, Watanabe T, Schlosser C, Merlini A, Körner H, Chang H-F, Fischer HJ, Reichardt HM, Zagrebelsky M, Mollenhauer B, Frahm J, Stadelmann C, Kügler S, Fitzner D, Haberl M, Odoardi F+ & Flügel A+ (2019) β Synuclein reactive T cells induce autoimmune CNS grey matter degeneration. Nature 566:503-508. *+equal contribution
  2. Flach A*, Litke T*, Strauss J*, Haberl M, Cordero Gómez C, Reindl M, Saiz A, Fehling HJ, Wienands J, Odoardi F, Lühder F, Flügel A (2016) Autoantibody-boosted T-cell reactivation in the target organ triggers manifestation of autoimmune CNS disease. Proc Natl Acad Sci USA 113: 3323-3328. *equal contribution
  3. Schläger C*, Körner H*, Krueger M, Vidoli S, Haberl M, Mielke D, Brylla E, Issekutz T, Cabañas C, Nelson PJ, Ziemssen T, Rohde V, Bechmann I, Lodygin D, Odoardi F*, Flügel A*. (2016) Effector T-cell trafficking between the leptomeninges and the cerebrospinal fluid. Nature 530:349-353. *equal contribution
  4. Lodygin D, Odoardi F, Schläger C, Körner H, Kitz A, Nosov M, van den Brandt J, Reichardt HM, Haberl M, Flügel A (2013) A combination of fluorescent NFAT and H2B sensors uncovers dynamics of T cell activation in real time during CNS autoimmunity. Nat Med 19:784-790.
  5. Odoardi F, Sie C, Streyl K, Ulaganathan VK, Schläger C, Lodygin D, Heckelsmiller K, Nietfeld W, Ellwart J, Klinkert WE, Lottaz C, Nosov M, Brinkmann V, Spang R, Lehrach H, Vingron M, Wekerle H, Flügel-Koch C, Flügel A (2012) T cells become licensed in the lung to enter the central nervous system. Nature 488:675-679.
  6. Bartholomäus I*, Kawakami N*, Odoardi F, Schläger C, Miljkovic D, Ellwart JW, Klinkert WE, Flügel-Koch C, Issekutz TB, Wekerle H, Flügel A (2009) Effector T cell interactions with meningeal vascular structures in nascent autoimmune CNS lesions. Nature 462:94-98. *equal contribution
  7. Odoardi F, Kawakami N, Klinkert WEF, Wekerle H, Flügel A (2007) Blood-born soluble protein antigen intensifies T cell activation in autoimmune CNS lesions and exacerbates clinical disease. Proc Natl Acad Sci USA 104:18625-18630.
  8. Kawakami N, Nägerl UV, Odoardi F, Bonhoeffer T, Wekerle H, Flügel A (2005) Live imaging of effector cell trafficking and autoantigen recognition within the unfolding autoimmune encephalomyelitis lesion. J Exp Med 201:1805-1814.
  9. Kawakami N, Lassmann S, Li Z, Odoardi F, Ritter T, Ziemssen T, Klinkert WE, Ellwart JW, Bradl M, Krivacic K, Lassmann H, Ransohoff RM, Volk HD, Wekerle H, Linington C, Flügel A (2004) The activation status of neuroantigen-specific T cells in the target organ determines the clinical outcome of autoimmune encephalomyelitis. J Exp Med 199:185-197.
  10. Flügel A, Berkowicz T, Ritter T, Labeur M, Jenne DE, Li Z, Ellwart JW, Willem M, Lassmann H, Wekerle H (2001) Migratory activity and functional changes of green fluorescent effector cells before and during experimental autoimmune encephalomyelitis. Immunity 14:547-560.

 

 

 

Our Team

Prof. Dr. Dr. Andreas H. Guse

Department of Biochemistry and Molecular Cell Biology

University Medical Center Hamburg-Eppendorf (UKE)

M.Sc. Imrankhan Khansahib

Department of Biochemistry and Molecular Cell Biology

University Medical Center Hamburg-Eppendorf (UKE)

M.Sc. Feng Gu

Department of Biochemistry and Molecular Cell Biology

University Medical Center Hamburg-Eppendorf (UKE)

M.Sc. Yunpeng Zhang

Department of Biochemistry and Molecular Cell Biology

University Medical Center Hamburg-Eppendorf (UKE)

Prof. Dr. Alexander Flügel

Institute of Neuroimmunology and Multiple Sclerosis Research
University Medical Center Göttingen (UMG)
 

Dr. Marc-André Lécuyer

Institute of Neuroimmunology and Multiple Sclerosis Research
University Medical Center Göttingen (UMG)
 

Dr. Henrike Körner

Institute of Neuroimmunology and Multiple Sclerosis Research
University Medical Center Göttingen (UMG)
 

Dr. Dmitri Lodygin

Institute of Neuroimmunology and Multiple Sclerosis Research
University Medical Center Göttingen (UMG)
 

Contact

University Medical Center Hamburg-Eppendorf 
Department of Biochemistry and Molecular Cell Biology 
Martinistrasse 52
20246 Hamburg

Scientific Coordinator

Dr. Björn-Philipp Diercks
Fon: +49  (0) 40 7410 54338
E-Mail: b.diercks©uke.de

Administration

Laura Mitsching
Fon: +49  (0) 40 7410 50301
E-Mail: l.mitsching©uke.de