AG Müller - Functional diversity of highly pathogenic Coronaviruses & comparative immune biology

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Research Focus

MERS- and SARS-Coronavirus (CoV) are highly pathogenic viruses associated with severe pneumonia and high mortality in humans. Both CoV likely originated in bats and were later transmitted to humans via other animal hosts (dromedaries, palm civets). We are exploring the phenotypic and functional diversity of MERS- and SARS-CoV using state-of-the-art cell culture models (air liquid interface epithelia, organoids) and reverse genetics. We aim to identify genetic markers that are responsible for human pathogenicity, virulence and transmissibility. Our research facilitates risk assessment of newly emerging CoV and increases our knowledge of virus evolution and virus-host adaptation.

Bats are important reservoir hosts for viruses, including CoV and seem to have developed an enhanced tolerance of viral infections. We are looking for differences between bats and humans regarding stress and immune response to understand the molecular basis of the differential outcomes of disease.

Our long-term goal is to characterize the basic mechanisms that facilitate animal-to-human virus transmission in order to pave the way for the development of new antiviral therapeutics.

Marcel A. Müller is "Highly Cited Researcher 2018" in the Cross-Field, Clarivate Analytics (https://hcr.clarivate.com/) category, making him one of the most cited researchers in his field (based on the evaluation of the Database "Web of Science")

Alumni: Dr. Daniela Niemeyer (now independent group leader), Dr. Benjamin Meyer, Dr. Susanne Biesold, Tasnim Suliman, PhD, Dipl.-Biol. Daniel Ritz, MD BSc Robert Wollny, Dr. Hanna Roth, MSc Schwiedhard Hoffstall, MSc Eva-Maria Klein, MSc Vincent Blaschke, 

Projects

  • Modell für die Verbreitung und Übertragung von MERS-Coronavirus in Endemiegebieten

    Modell für die Verbreitung und Übertragung von MERS-Coronavirus in Endemiegebieten Image /

  • Phylogenetic analysis of currently known MERS-Coronavirus clades and their distribution in Africa and the Arabian Peninsula.

    Phylogenetic analysis of currently known MERS-Coronavirus clades and their distribution in Africa and the Arabian Peninsula. Image /

  • Our research projects focus on the characterization of virus- and host-specific restriction factors that facilitate virus transmission from animals to humans.

    Our research projects focus on the characterization of virus- and host-specific restriction factors that facilitate virus transmission from animals to humans. Image /

  • In the renovated and recently re-opened Biosafety Level 3 (BSL3) facility we work with MERS- and SARS-Coronavirus.

    In the renovated and recently re-opened Biosafety Level 3 (BSL3) facility we work with MERS- and SARS-Coronavirus. Image /

  • In the renovated and recently re-opened Biosafety Level 3 (BSL3) facility we work with MERS- and SARS-Coronavirus.

    In the renovated and recently re-opened Biosafety Level 3 (BSL3) facility we work with MERS- and SARS-Coronavirus. Image /

Evolution and functional diversity of MERS coronavirus

MERS-CoV is widespread in dromedaries across Africa and the Arabian Peninsula and can cause a lethal form of pneumonia in humans. There are multiple genetic polymorphisms in circulating MERS-CoV strains. We want to explore if there are phenotypical differences due to the observed genotypic diversity of MERS-CoV in dromedaries and humans. By using in vitro cell culture models we aim at identifying genetic markers that correlate with increased pathogenicity and that can be used for risk assessment of newly emerging MERS-CoV strains. Finally, the established reverse genetics system for MERS-CoV will allow the identification of molecular mechanisms responsible for putative phenotypes in cell culture models.

Funding: RAPID (PI Drosten), National Science Foundation (PIs Müller, Drosten)

Contact: PD Dr. Marcel Müller, MSc Simon Schroeder

Immunological features of virus infections in bats

Bats carry a plethora of viruses without showing overt clinical symptoms. We hypothesize that the virus-host interplay in bats results in less immunological and proinflammatory activation compared to humans. In addition, bats may be more resistant towards external stressors. By using newly established cell culture-based in vitro assays (real time PCRs, interferon promoter activation reporter assays and interferon bioassay) for bats and humans, we aim to analyze and directly compare the basic features of the interferon and stress response of bats. A combination of virus infections and different stressors/immune stimuli will be examinend in order to understand the interplay between immune activation and virus replication and to identify differences between bat and human cells.

Funding: Volkswagen Stiftung (PI Müller)

Contact: PD Dr. Marcel Müller, MSc Jan Papies

Identification and characterization of bat-specific nucleic acid sensor proteins

Bats are less prone to the development of cancer and pathogen-related diseases, which may be a result of unique features in their innate immune response to foreign or damaged nucleic acids. We hypothesize that bats have other, still unknown, DNA sensors that are differentially activated by foreign or damaged DNA molecules. Alternatively, bats may have novel or differentially active nucleic acid sensors or second messenger molecules that can compensate for the absence of DNA sensors.

Bat-specific nucleic acid-binding proteins will be identified by proteomics and protein pull down approaches followed by mass spectrometry (in collaboration with Prof. Andreas Pichlmair, Technical University Munich). Putative bat-specific nucleic acid sensors will be further characterized by siRNA-based knockdowns and CRISPR/Cas-based knockouts of identified target genes. The effects of target knockdowns/knockouts on the innate immune response of bat cell cultures will be assessed by bat-specific real time PCRs and promoter reporter assays as well as newly established CRISPR/Cas-based reporter cell lines. Finally, gain-of-function experiments and co-immunoprecipitations will be used to confirm protein-nucleic acid interactions.

Funding: Volkswagen Stiftung (PI Müller)

Contact: PD Dr. Marcel Müller, MSc Andrea Sieberg

Methods

  • Working under BSL2 and BSL3 conditions
  • primary and immortalized cell cultures
  • CRISPR/Cas9-based generation of knock-out or knock-in reporter cell lines
  • luciferase promoter activation assays
  • transcriptome and proteome analyses (e.g. real-time PCR, Interferon bioassays, ELISA, Immunofluorescence assays)
  • Live-cell imaging (LSM800 Airyscan),
  • FACS,
  • Immunoprecipitations
  • SDS-PAGE and Western blotting

External collaborators

  • Dr. Nils Gassen, University of Bonn Medical Centre, Institute of Psychiatry, Bonn
  • Dr. Stella Kiambi, Food and Agriculture Organization (FAO), Kenya
  • Prof. Andreas Pichlmair, Technical University Munich (TUM), Munich
  • Prof. Amira Roess, University of Washington, Washington DC, USA
  • Prof. Friedemann Weber, University of Gießen
  • Prof. Georg Kochs, University of Freiburg
  • Prof. Volker Thiel, Dr. Ronald Dijkman, Bern, Switzerland
  • Prof. Stefan Pöhlmann, Dr. Markus Hoffmann, DPZ, Göttingen
  • Prof. Susan Baker, Loyola University, Chicago, USA
  • Prof. Alexander Lukashev, Sechenov University, Moscow, Russia

Significant publications

A current list of all publications can be found in the pubmed-List or on

Google Scholar.

 

5 important publications (*shared authorships)

1. Gassen NC, Niemeyer D, Muth D, Corman VM, Martinelli S, Gassen A, Hafner K, Papies J, Mösbauer K, Zellner A, Zannas AS, Herrmann A, Holsboer F, Brack-Werner R, Boshart M, Müller-Myhsok B, Drosten C, Müller MA*, Rein T*. (2019) SKP2 attenuates autophagy through Beclin1-ubiquitination and its inhibition reduces MERS-Coronavirus infection. Nat Commun 10(1):5770.

LINK: https://www.nature.com/articles/s41467-019-13659-4

2.     Kiambi S*, Corman VM*, Sitawa R, Githinji J, Ngoci J, Ozomata AS, Gardner E, von Dobschuetz S, Morzaria S, Kimutai J, Schroeder S, Njagi O, Simpkin P, Rugalema G, Tadesse Z, Lubroth J, Makonnen Y, Drosten C, Muller MA*, Fasina FO*. (2018) Detection of distinct MERS-Coronavirus strains in dromedary camels from Kenya, 2017. Emerg Microbes Infect 7:195.

LINK: https://www.tandfonline.com/doi/full/10.1038/s41426-018-0193-z 

3.  Niemeyer D, Mosbauer K, Klein EM, Sieberg A, Mettelman RC, Mielech AM, Dijkman R, Baker SC, Drosten C*, Muller MA*. (2018) The papain-like protease determines a virulence trait that varies among members of the SARS-coronavirus species. PLoS Pathog 14:e1007296.

LINK: https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1007296&type=printable

4. Muller MA, Meyer B, Corman VM, Al-Masri M, Turkestani A, Ritz D, Sieberg A, Aldabbagh S, Bosch BJ, Lattwein E, Alhakeem RF, Assiri AM, Albarrak AM, Al-Shangiti AM, Al-Tawfiq JA, Wikramaratna P, Alrabeeah AA, Drosten C*, Memish ZA*. (2015) Presence of Middle East respiratory syndrome coronavirus antibodies in Saudi Arabia: a nationwide, cross-sectional, serological study. The Lancet infectious diseases 15(5):559-564.

LINK: https://www.sciencedirect.com/science/article/pii/S1473309915700903?via%3Dihub

5.  Reusken CB*, Haagmans BL*, Muller MA*, Gutierrez C, Godeke GJ, Meyer B, Muth D, Raj VS, Vries LS, Corman VM, Drexler JF, Smits SL, El Tahir YE, De Sousa R, van Beek J, Nowotny N, van Maanen K, Hidalgo-Hermoso E, Bosch BJ, Rottier P, Osterhaus A, Gortazar-Schmidt C, Drosten C, Koopmans MP. (2013) Middle East respiratory syndrome coronavirus neutralising serum antibodies in dromedary camels: a comparative serological study. The Lancet infectious diseases 13:859-866.

LINK: https://www.thelancet.com/action/showPdf?pii=S1473-3099%2813%2970164-6