A18

A18: Summary

SARS-CoV-2, a novel corona virus is responsible for the current pandemic of Covid-19, a novel form of pneumonia which can have life-threatening complications like ARDS, hypercoagulation and multi organ failure. nsp3, one of the non-structural proteins of SARS-CoV-2 contains an ADPR-binding macroD-like macrodomain. The homologous macrodomain of related viruses exhibits catalytic activity, removing ADPR from mono- or poly-ADP-ribosylated proteins (deMARylation/dePARylation enzyme activity). Post-translational modifications by ADP-ribosylation are involved in the interferon response to RNA viruses in infected cells and the subsequent induction of interferon-stimulated genes in neighbouring cells that result in an antiviral state and limit viral spread in the tissue. The deMARylation/dePARylation of proteins in these pathways might therefore contribute to the observed immune evasion by the virus, which allows for viral replication with the high virus titers promoting inflammation and disease progression. In the proposed project we intend to investigate the deMARylation/dePARylation activity of nsp3 as a potential target for a novel anti-viral approach that might prevent this suppression of the interferon response by the virus. We will develop biochemical assays for the hydrolase activity of nsp3 and investigate its structure activity relationship (SAR) using ADPR analogues. In addition, non-nucleotide inhibitors will be developed by virtual screening using a structure-based approach based on available crystal structures in the PDB and a ligandbased approach based on our SAR data. Compounds will be tested in biochemical assays and in cellular assays with regard to their activity towards the interferon response to RNA intermediates. Compounds will be tested in biochemical assays and in cellular assays with regard to their activity towards the interferon response to synthetic double-stranded or hairpin RNA. Compounds that proved to be active in the biochemical and cellular assays will be used to explore the role of the enzymatic activity of the viral macrodomain for viral replication, interferon response and interferon sensitivity in cells infected with SARS-CoV-2.

Dr. Ralf Fliegert

Selected publications (*equal contribution)

 

1. Fliegert R, Riekehr WM, Guse AH (2020) Does Cyclic ADP-Ribose (cADPR) Activate the Non-selective Cation Channel TRPM2?. Front Immunol 11:2018
2. Baszczyňski O, Watt JM, Rozewitz MD, Guse AH, Fliegert R, Potter BVL (2019) Synthesis of Terminal Ribose Analogues of Adenosine 5'-Diphosphate Ribose as Probes for the Transient Receptor Potential Cation Channel TRPM2. J Org Chem  84:6143-6157
3. Fliegert R*, Heeren J*, Koch-Nolte F*, Nikolaev VO*, Lohr C*, Meier C*, Guse AH* (2019) Adenine nucleotides as paracrine mediators and intracellular second messengers in immunity and inflammation. Biochem Soc Trans 47(1):329-337.
4. Gattkowski E, Johnsen A, Bauche A, Möckl F, Kulow F, Garcia Alai M, Rutherford TJ, Fliegert R*, Tidow H* (2019) Novel CaM-binding motif in its NudT9H domain contributes to temperature sensitivity of TRPM2. Biochim Biophys Acta Mol Cell Res 66(7):1162-1170.
5. 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 Biol13:10361044.
6. Fliegert R, Watt JM, Schöbel A, Rozewitz MD, Moreau C, Kirchberger T, Thomas MP, Sick W, Araujo AC, Harneit A, Potter BVL, Guse AH (2017) Ligand-induced activation of human TRPM2 requires the terminal ribose of ADPR and involves Arg1433 and Tyr1349. Biochem J474:2159-2175.
7. Guse AH, Ernst IM, Fliegert R (2013) NAADP signaling revisited. Curr Top Med Chem 13:29782990.
8. Schwarz N*, Fliegert R*, Adriouch S*, Seman M, Guse AH, Haag F, Koch-Nolte F (2009) Activation of the P2X7 ion channel by soluble and covalently bound ligands. Purinergic Signal 5:139-149.
9. Fliegert R, Gasser A, Guse AH (2007) Regulation of calcium signalling by adenine-based second messengers. Biochem Soc Trans 35:109-114.
10. Fliegert R, Glassmeier G, Schmid F, Cornils K, Genisyuerek S, Harneit A, Schwarz JR, Guse AH (2007) Modulation of Ca2+ entry and plasma membrane potential by human TRPM4b. FEBS J 274:704713.

Prof. Dr. Chris Meier

Selected publications

 

1. Zhao C, Weber S, Schols D, Balzarini J, Meier C (2020) Prodrugs of γ-Alkyl-Modified Nucleoside Triphosphates: Improved Inhibition of HIV Reverse Transcriptase. Angew Chem Int Ed Engl 59:22063-22071
2. Jia X, Schols D, Meier C (2020) Lipophilic Triphosphate Prodrugs of Various Nucleoside Analogues. J Med Chem 63:6991-7007.
3. Jia X, Schols D, Meier C (2020) Anti-HIV-active Nucleoside Triphosphate Prodrugs. J Med Chem 63:6003-27.
4. Fehling H*, Choy SL*, Ting F*, Landschulze D, Bernin H, Lender SC, Mühlenpfordt M, Bifeld E, Eick J, Marggraff C, Kottmayr N, Groneberg M, Hoenow S, Sellau J, Clos J, Meier C, Lotter H  (2020) Antileishmanial Effects of Synthetic EhPIb Analogs Derived from the Entamoeba histolytica Lipopeptidephosphoglycan. Antimicrob Agents Chemother 64:e00161-20. 
5. Shannon A, Selisko B, Le NT, Huchting J, Touret F, Piorkowski G, Fattorini V, Ferron F, Decroly E, Meier C, Coutard B, Peersen O, Canard B (2020) Rapid incorporation of Favipiravir by the fast and permissive viral RNA polymerase complex results in SARS-CoV-2 lethal mutagenesis. Nature Commun 11:4682.
6. Huchting J, Vanderlinden E, Winkler M, Nasser H, Naesens L, Meier C (2018) Prodrugs of the Phosphoribosylated Forms of Hydroxypyrazinecarboxamide Pseudobase T-705 and its De-Fluoro-Analogue T-1105 as Potent Influenza Virus Inhibitors. J Med Chem 61:6193-210.
7. Diercks BP, Werner R, Weidemüller P, Czarniak F, Hernandez L, Lehmann C, Rosche A, Krüger A, Kaufmann U, Vaeth M, Failla AV, Zobiak B, Kandil FI, Schetelig D, Ruthenbeck A, Meier C, Lodygin D, Flügel A, Ren D, Wolf IMA, Feske S, Guse AH (2018) ORAI1, STIM1/2, and RYR1 shape subsecond Ca²⁺ microdomains upon T cell activation. Sci Signal 18;11(561).
8. Gollnest T, Dinis de Oliveira T, Rath A, Hauber I, Schols D, Balzarini J, Meier C (2016) Membrane-permeable Triphosphate Prodrugs of Nucleoside Analogues. Angew Chem Int Ed 55:5255-5258.
9. Gollnest T, Dinis de Oliveira T, Schols D, Balzarini J, Meier C (2015) The TriPPPro-Approach: Lipophilic Prodrugs of Nucleoside Triphosphates as Biochemical Probes and Potential Antivirals. Nat Commun 6:8716
10. Weinschenk L, Schols D, Balzarini J, Meier C (2015) Nucleoside Diphosphate Prodrugs: Nonsymmetric DiPPro-Nucleotides. J Med Chem 58:6114-6130.

Dr. Susanne Pfefferle

Selected publications (*equal contribution)

 

1. Pfefferle S*, Gunther T*, Kobbe R*, Czech-Sioli M, Norz D, Santer R, Oh J, Kluge S, Oestereich L, Peldschus K, Indenbirken D, Huang J, Grundhoff A, Aepfelbacher M, Knobloch JK, Lutgehetmann M, Fischer N (2021) SARS Coronavirus-2 variant tracing within the first Coronavirus Disease 19 clusters in northern Germany. Clin Microbiol Infect 27(1): p. 130 e5-130 e8.
2. Norz D, Fischer N, Schultze A, Kluge S, Mayer-Runge U, Aepfelbacher M, Pfefferle S*, Lutgehetmann M* (2020) Clinical evaluation of a SARS-CoV-2 RT-PCR assay on a fully automated system for rapid on-demand testing in the hospital setting. J Clin Virol 128: p. 104390.
3. Heinrich F, Meissner K, Langenwalder F, Puschel K, Norz D, Hoffmann A, Lutgehetmann M, Aepfelbacher M, Bibiza-Freiwald E, Pfefferle S*, Heinemann A* (2021) Postmortem Stability of SARS-CoV-2 in Nasopharyngeal Mucosa. Emerg Infect Dis 27(1).
4. Zhao Y, Kilian C, Turner JE, Bosurgi L, Roedl K, Bartsch P, Gnirck AC, Cortesi F, Schultheiss C, Hellmig M, Enk LUB, Hausmann F, Borchers A, Wong MN, Paust HJ, Siracusa F, Scheibel N, Herrmann M, Rosati E, Bacher P, Kylies D, Jarczak D, Lutgehetmann M, Pfefferle S, Steurer S, Schulze-Zur-Wiesch J, Puelles VG, Sperhake JP, Addo MM, Lohse AW, Binder M, Huber S, Huber TB, Kluge S, Bonn S, Panzer U, Gagliani N, Krebs CF (2021) Clonal expansion and activation of tissue-resident memory-like Th17 cells expressing GM-CSF in the lungs of severe COVID-19 patients. Science Immunol 6(56).
5. Braun F*, Lutgehetmann M*, Pfefferle S*, Wong MN, Carsten A, Lindenmeyer MT, Norz D, Heinrich F, Meissner K, Wichmann D, Kluge S, Gross O, Pueschel K, Schroder AS, Edler C, Aepfelbacher M, Puelles VG, Huber TB (2020) SARS-CoV-2 renal tropism associates with acute kidney injury. Lancet 396(10251): p. 597-598.
6. Norz D, Frontzek A, Eigner U, Oestereich L, Wichmann D, Kluge S, Fischer N, Aepfelbacher M, Pfefferle S*, Lutgehetmann M* (2020) Pushing beyond specifications: Evaluation of linearity and clinical performance of the cobas 6800/8800 SARS-CoV-2 RT-PCR assay for reliable quantification in blood and other materials outside recommendations. J Clin Virol 132: p. 104650.
7. Wichmann D, Sperhake JP, Lutgehetmann M, Steurer S, Edler C, Heinemann A, Heinrich F, Mushumba H, Kniep I, Schroder AS, Burdelski C, de Heer G, Nierhaus A, Frings D, Pfefferle S, Becker H, Bredereke-Wiedling H, de Weerth A, Paschen HR, Sheikhzadeh-Eggers S, Stang A, Schmiedel S, Bokemeyer C, Addo MM, Aepfelbacher M, Puschel K, Kluge S (2020) Autopsy Findings and Venous Thromboembolism in Patients With COVID-19: A Prospective Cohort Study. Ann Intern Med 173(4):268-277
8. Puelles VG, Lutgehetmann M, Lindenmeyer MT, Sperhake JP, Wong MN, Allweiss L, Chilla S, Heinemann A, Wanner N, Liu S, Braun F, Lu S, Pfefferle S, Schroder AS, Edler C, Gross O, Glatzel M, Wichmann D, Wiech T, Kluge S, Pueschel K, Aepfelbacher M, Huber TB (2020) Multiorgan and Renal Tropism of SARS-CoV-2. N Engl J Med 383:590-592
9. Pfefferle S, SchopfJ, Kogl M, Friedel CC, Muller MA, Carbajo-Lozoya J, Stellberger T, von Dall'Armi E, Herzog P, Kallies S, Niemeyer D, Ditt V, Kuri T, Zust R, Pumpor K, Hilgenfeld R, Schwarz F, Zimmer R, Steffen I, Weber F, Thiel V, Herrler G, Thiel HJ, Schwegmann-Wessels C, Pohlmann S, Haas J, Drosten C, von Brunn A (2011) The SARS-coronavirus-host interactome: identification of cyclophilins as target for pan-coronavirus inhibitors. PLoS Pathog 7(10): p. e1002331.
10. Pfefferle S, Krahling V, Ditt V, Grywna K, Muhlberger E, Drosten C (2009) Reverse genetic characterization of the natural genomic deletion in SARS-Coronavirus strain Frankfurt-1 open reading frame 7b reveals an attenuating function of the 7b protein in-vitro and in-vivo. Virol J 6: p. 131.

Our Team