A18

A18: Summary

As part of non-structural protein 3 (nsp3) SARS-CoV-2 harbours an ADPR-binding macrodomain with deADP-ribosylating activity that contributes to the suppression of the antiviral interferon response of the host. Aim of the project is to develop a structure-activity relationship for the viral macrodomain using ADPR analogues and to identify non-nucleotide inhibitors using a virtual screening approach. In addition we will analyse the role of the macrodomain in infection experiments using a reverse genetics approach and determine the effect of naturally occurring mutations in the macrodomain of nsp3 on enzymatic activity and viral replication.

Dr. Ralf Fliegert

Selected publications (*equal contribution)

 

1. Sandmann M, Tajdar S, Sander S, CarrilloD, Ganter B, Ocenas M, Etzold S, Pekarek N, Berger J, Meister TL, Watt J, Baszczyňski O, Potter BVL, Alai MG, Tidow H, Pfefferle S, Meier C, Fliegert R (2025) Targeting a viral macrodomain: Design, Structure-Based Optimization and Antiviral Evaluation of Nanomolar Inhibitors for Mac1 of SARS-CoV-2. Preprint at Research Square https://doi.org/10.21203/rs.3.rs-7063434/v1
2. Uebler L, Sandmann M, Pekarek N, Berger J, Lütgehetmann M, von Possel R, Emmerich P, Fliegert R, Pfefferle S (2025) Generation and Characterization of a Tagged Recombinant SARS‐CoV‐2 for Functional Replicase Studies. Preprint at Preprints.org DOI:10.20944/preprints202510.0092.v1
3. Fliegert R, Riekehr WM, Guse AH (2020) Does Cyclic ADP-Ribose (cADPR) Activate the Non-selective Cation Channel TRPM2?. Front Immunol 11:2018
4. 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
5. 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.
6. 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.
7. 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.
8. 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.
9. Guse AH, Ernst IM, Fliegert R (2013) NAADP signaling revisited. Curr Top Med Chem 13:29782990.
10. 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.

Prof. Dr. Chris Meier

Selected publications

 

1. Winterberg KJ, Schwentner V, Gu F, Möckl F, Li G, Bauche A, Etzold S, Rosche A, Weiß M, Thuille N, Förster F, Woelk L, Werner R, Kovacevic D, Fehse B, Kurelic R, Nawrocki M, Huber S, Mittrücker HW, Meier C, Müller CE, Baier G, Skålhegg BS, de Deken X, Wahl-Schott C, Mair T, Siebels B, Cugota Canals R, Odoardi F, Lodygin D, Flügel A, Nikolaev VO, Diercks BP, Guse AH. Multiple signaling events are required for NAADP synthesis by DUOX2 and formation of Ca²⁺ microdomains to initiate T cell activation. Sci Signal. 2026 Jan 13;19(920):eadp4326.
2. Nikolova V, Linnemannstöns K, Bendel ME, Machado M, Ganter B, Budimir P, Vogts M, Fischer C, Ganter M, Meier C, Dobbelstein M. Membrane-Permeable 5-Fluorodeoxyuridine Triphosphate Derivatives Inhibit the Proliferation of Plasmodium falciparum. ACS Infect Dis. 2025 Nov 14;11(11):3138-3151.
3. Knaack JIH, List E, Stalling D, Sterrenberg VT, Meier C, Wagenknecht HA, Bosse JB. Live-Cell RNA Imaging via Clickable TriPPPro Nucleotide Reporters. Angew Chem Int Ed Engl. 2026 Jan 9;65(2):e16613.
4. Vogts M, Witt J, Riedner M, Meier C. Development of a HILIC-MS/MS method for simultaneous quantification of lipophilic antitumor TriPPPro-prodrugs and their polar metabolites in HT29 cell extracts. J Chromatogr B Analyt Technol Biomed Life Sci. 2025 Sep 1;1263:124673.
5. Krukenberg S, Möckl F, Weiß M, Dekiert P, Hofmann M, Gerlach F, Winterberg KJ, Kovacevic D, Khansahib I, Troost B, Hinrichs M, Granato V, Nawrocki M, Hub T, Tsvilovskyy V, Medert R, Woelk LM, Förster F, Li H, Werner R, Altfeld M, Huber S, Clarke OB, Freichel M, Diercks BP, Meier C*, Guse AH*. MASTER-NAADP: a membrane permeable precursor of the Ca²⁺mobilizing second messenger NAADP. Nat Commun. 2024 Sep 13;15(1):8008.
6. Sterrenberg VT, Stalling D, Knaack JIH, Soh TK, Bosse JB, Meier C. A TriPPPro-Nucleotide Reporter with Optimized Cell-Permeable Dyes for Metabolic Labeling of Cellular and Viral DNA in Living Cells. Angew Chem Int Ed Engl. 2023 Sep 18;62(38):e202308271. 
7. Nawrocki M, Lory N, Bedke T, Stumme F, Diercks BP, Guse AH, Meier C, Gagliani N, Mittrücker HW, Huber S. Trans-Ned 19-Mediated Antagonism of Nicotinic Acid Adenine Nucleotide-Mediated Calcium Signaling Regulates Th17 Cell Plasticity in Mice. Cells. 2021 Nov 5;10(11):3039.
8. 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. Dual NADPH oxidases DUOX1 and DUOX2 synthesize NAADP and are necessary for Ca²⁺ signaling during T cell activation. Sci Signal. 2021 Nov 16;14(709):eabe3800. 
9. 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
10. Jia X, Schols D, Meier C (2020) Lipophilic Triphosphate Prodrugs of Various Nucleoside Analogues. J Med Chem 63:6991-7007.

PD Dr. Susanne Pfefferle

Selected publications (*equal contribution)

 

1. Pfefferle S, Nörz D, Giersch K, Grunwald M, Tang HT, Pflüger LS, et al. Refined fully automated RT-qPCR assay for simultaneous detection of SARS-CoV-2, Influenza A/B, and RSV with target optimization for improved variant resilience. Journal of Clinical Virology 2026;183:105924
2. Sandmann M, Tajdar S, Sander S, CarrilloD, Ganter B, Ocenas M, Etzold S, Pekarek N, Berger J, Meister TL, Watt J, Baszczyňski O, Potter BVL, Alai MG, Tidow H, Pfefferle S, Meier C, Fliegert R (2025) Targeting a viral macrodomain: Design, Structure-Based Optimization and Antiviral Evaluation of Nanomolar Inhibitors for Mac1 of SARS-CoV-2. Preprint at Research Square https://doi.org/10.21203/rs.3.rs-7063434/v1
3. Uebler L, Sandmann M, Pekarek N, Berger J, Lütgehetmann M, von Possel R, Emmerich P, Fliegert R, Pfefferle S (2025) Generation and Characterization of a Tagged Recombinant SARS‐CoV‐2 for Functional Replicase Studies. Preprint at Preprints.org DOI:10.20944/preprints202510.0092.v1
4. 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.
5. 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.
6. 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).
7. 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).
8. 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.
9. 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.
10. 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

Our Team