Viral tools and monosynaptic tracers for studying SARS-CoV2 neurotropism and neuronal transmission
Molecular and Cellular Neurology and Neuropathology
Final Report Abstract
Neurological deficiencies in COVID-19 patients like loss of smell and taste prompted the idea of neurotropism of SARS-CoV-2. In this one year COVID-19 Focus Funding project, rabies virus (RABV) and vesicular stomatitis virus (VSV) pseudotype particles carrying spike proteins (S) of SARS-CoV-2 and variants of concern (VOCs) were generated to study the permissivity of iPSC-derived neurons and astrocytes. S constructs with modifications of the cytoplasmic tail remained fusion active, as determined in cell-cell fusion assays, but were poorly incorporated into pseudotype virus particles encoding the reporter genes Gaussia luciferase and/or eGFP, and yielded moderate infectious titers. Highest infectivity with titers of 104-105/mLwas observed in VeroE6 and Caco2 cells, while iPSC-derived human astrocytes and neurons were even less permissive than negative controls. Parallel experiments with G proteins from novel potentially zoonotic lyssaviruses from amphibians and reptiles (nLy-19) yielded 100 fold higher titers. Due to the poor infectivity of S pseudotype viruses for human neurons the envisaged transsynaptic tracing experiments in transgenic hACE2 mice were not justified. Neutralization assays established in the project further served as controls for the integrity and antigenicity of the pseudotype viruses carrying S from Wuhan virus and VOCS. Sera from COVID-19 patients and vaccinees, or immunized mice as well as monoclonal antibodies confirmed comparable behavior of pseudotype (BSL1) and SARS-CoV-2 viruses (BSL3). Virus aliquots were provided to several other groups and for collaborations.
Publications
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Glycoproteins of Predicted Amphibian and Reptile Lyssaviruses Can Mediate Infection of Mammalian and Reptile Cells. Viruses, 13(9), 1726.
Oberhuber, Martina; Schopf, Anika; Hennrich, Alexandru Adrian; Santos-Mandujano, Rosalía; Huhn, Anna Gesine; Seitz, Stefan; Riedel, Christiane & Conzelmann, Karl-Klaus
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Optimization of whole-brain rabies virus tracing technology for small cell populations. Scientific Reports, 11(1).
Roelofs, Theresia J. M.; Menting-Henry, Shanice; Gol, Lieke M.; Speel, Annelijn M.; Wielenga, Vera H.; Garner, Keith M.; Luijendijk, Mieneke C. M.; Hennrich, Alexandru A.; Conzelmann, Karl-Klaus & Adan, Roger A. H.
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Reprogramming reactive glia into interneurons reduces chronic seizure activity in a mouse model of mesial temporal lobe epilepsy. Cell Stem Cell, 28(12), 2104-2121.e10.
Lentini, Célia; d.’Orange, Marie; Marichal, Nicolás; Trottmann, Marie-Madeleine; Vignoles, Rory; Foucault, Louis; Verrier, Charlotte; Massera, Céline; Raineteau, Olivier; Conzelmann, Karl-Klaus; Rival-Gervier, Sylvie; Depaulis, Antoine; Berninger, Benedikt & Heinrich, Christophe
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Safe and effective two-in-one replicon-and-VLP minispike vaccine for COVID-19: Protection of mice after a single immunization. PLOS Pathogens, 17(4), e1009064.
Hennrich, Alexandru A.; Sawatsky, Bevan; Santos-Mandujano, Rosalía; Banda, Dominic H.; Oberhuber, Martina; Schopf, Anika; Pfaffinger, Verena; Wittwer, Kevin; Riedel, Christiane; Pfaller, Christian K. & Conzelmann, Karl-Klaus
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Spike residue 403 affects binding of coronavirus spikes to human ACE2. Nature Communications, 12(1).
Zech, Fabian; Schniertshauer, Daniel; Jung, Christoph; Herrmann, Alexandra; Cordsmeier, Arne; Xie, Qinya; Nchioua, Rayhane; Prelli, Bozzo Caterina; Volcic, Meta; Koepke, Lennart; Müller, Janis A.; Krüger, Jana; Heller, Sandra; Stenger, Steffen; Hoffmann, Markus; Pöhlmann, Stefan; Kleger, Alexander; Jacob, Timo; Conzelmann, Karl-Klaus ... & Kirchhoff, Frank
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Systematic functional analysis of SARS-CoV-2 proteins uncovers viral innate immune antagonists and remaining vulnerabilities. Cell Reports, 35(7), 109126.
Hayn, Manuel; Hirschenberger, Maximilian; Koepke, Lennart; Nchioua, Rayhane; Straub, Jan Hendrik; Klute, Susanne; Hunszinger, Victoria; Zech, Fabian; Prelli, Bozzo Caterina; Aftab, Wasim; Christensen, Maria Hønholt; Conzelmann, Carina; Müller, Janis Alexander; Srinivasachar, Badarinarayan Smitha; Stürzel, Christina Martina; Forne, Ignasi; Stenger, Steffen; Conzelmann, Karl-Klaus; Münch, Jan ... & Sparrer, Konstantin Maria Johannes
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Excessive local host-graft connectivity in aging and amyloid-loaded brain. Science Advances, 8(23).
Thomas, Judith; Martinez-Reza, Maria Fernanda; Thorwirth, Manja; Zarb, Yvette; Conzelmann, Karl-Klaus; Hauck, Stefanie M.; Grade, Sofia & Götz, Magdalena
