Project Description

TP18: Combinatorial knock-down/knock-out strategies to reconstitute anti-HBV immune responses and to eliminate persisting HBV cccDNA

Our aim is to exploit potent technologies for gene knock-down/knock-out to target Hepatitis B virus (HBV) gene expression and persistence in immunocompetent mice, in order to study the ensuing effects on the antiviral immune response and control of viral infection. To this end, we will develop and use novel Adeno-associated viral gene transfer vectors that co-deliver anti-HBV RNAi and CRISPR expression cassettes to HBV-infected hepatocytes. These vectors hold great potential as tools for basic studies into hepatitis virus biology and persistence also by other TRR179 members as well as for translation into a clinical modality.
Hoffmann MD, Aschenbrenner S, Grosse S, Rapti K, Domenger C, Fakhiri J, Mastel M, Börner K, Eils R, Grimm D, Niopek D. 2019. Cell-specific CRISPR-Cas9 activation by microRNA-dependent expression of anti-CRISPR proteins. Nucleic Acids Res. 47(13):e75. doi: 10.1093/nar/gkz271
Bubeck F, Hoffmann MD, Harteveld Z, Aschenbrenner S, Bietz A, Waldhauer MC, Börner K, Fakhiri J, Schmelas C, Dietz L, Grimm D, Correia BE, Eils R, Niopek D. 2018. Engineered anti-CRISPR proteins for optogenetic control of CRISPR-Cas9. Nat Methods. 15(11):924-927
Schmelas C, Grimm D. 2018. Split Cas9, Not Hairs - Advancing the Therapeutic Index of CRISPR Technology. Biotechnol J. 13(9):e1700432. doi: 10.1002/biot.201700432. Review.
Herrmann AK, Grimm D. 2018. High-Throughput Dissection of AAV-Host Interactions: The Fast and the Curious. J Mol Biol. 430(17):2626-2640. doi: 10.1016/j.jmb.2018.05.022. Review.
Senís E, Mosteiro L, Wilkening S, Wiedtke E, Nowrouzi A, Afzal S, Fronza R, Landerer H, Abad M, Niopek D, Schmidt M, Serrano M, Grimm D. 2018. AAVvector-mediated in vivo reprogramming into pluripotency. Nat Commun. 9(1):2651. doi: 10.1038/s41467-018-05059-x.
Schiwon M, Ehrke-Schulz E, Oswald A, Bergmann T, Michler T, Protzer U, Ehrhardt A. 2018. One-Vector System for Multiplexed CRISPR/Cas9 against Hepatitis B Virus cccDNA Utilizing High-Capacity Adenoviral Vectors. Mol Ther Nucleic Acids. 12:242-253. doi: 10.1016/j.omtn.2018.05.006.
Schmelas C, Grimm D. 2018. Split Cas9, Not Hairs - Advancing the Therapeutic Index of CRISPR Technology. Biotechnol J. 2018 Jan 5. doi: 10.1002/biot.201700432. (Epub ahead of print) Review.
Lempp FA, Wiedtke E, Qu B, Roques P, Chemin I, Vondran FW, Le Grand R, Grimm D, Urban S. 2017. Sodium taurocholate cotransporting polypeptide is the limiting host factor of Hepatitis B Virus infection in macaque and pig hepatocytes. Hepatology. doi: 10.1002/hep.29112.
Senís E, Mockenhaupt S, Rupp D, Bauer T, Paramasivam N, Knapp B, Gronych J, Grosse S, Windisch MP, Schmidt F, Theis FJ, Eils R, Lichter P, Schlesner M, Bartenschlager R, Grimm D. 2017. TALEN/CRISPR-mediated engineering of a promoterless anti-viral RNAi hairpin into an endogenous miRNA locus. Nucleic Acids Res. 45(1):e3. doi: 10.1093/nar/gkw805.
Michler T, Große S, Mockenhaupt S, Röder N, Stückler F, Knapp B, Ko C, Heikenwalder M, Protzer U, Grimm D. 2016. Blocking sense-strand activity improves potency, safety and specificity of anti-hepatitis B virus short hairpin RNA. EMBO Mol Med. 8(9):1082-98. doi: 10.15252/emmm.201506172.
Mockenhaupt S, Grosse S, Rupp D, Bartenschlager R, Grimm D. 2015. Alleviation of off-target effects from vector-encoded shRNAs via codelivered RNA decoys. Proc Natl Acad Sci U S A 112:E4007-16.
Senís E, Fatouros C, Große S, Wiedtke E, Niopek D, Mueller AK, Börner K, Grimm D. 2014. CRISPR/Cas9-mediated genome engineering: An adenoassociated viral (AAV) vector toolbox. Biotechnol J. 9:1402-12.
Hösel M, Lucifora J, Michler T, Holz G, Gruffaz M, Stahnke S, Zoulim F, Durantel D, Heikenwalder M, Nierhoff D, Millet R, Salvetti A, Protzer U, Büning H.. 2014. Hepatitis B Virus enhances efficacy of Adeno- Associated Viral vectors in vitro and in vivo. Hepatology. 59:2110-20.
Börner K, Niopek D, Cotugno G, Kaldenbach M, Pankert T, Willemsen J, Zhang X, Schürmann N, Mockenhaupt S, Serva A, Hiet MS, Wiedtke E, Castoldi M, Starkuviene V, Erfle H, Gilbert DF, Barten- schlager R, Boutros M, Binder M, Streetz K, Kräusslich HG, Grimm D. 2013. Robust RNAi enhancement via human Argonaute-2 over-expression from plasmids, viral vectors and cell lines. Nucleic Acids Re- search 41:e199.
Huang LR, Wohlleber D, Reisinger F, Jenne CN, Cheng RL, Abdullah Z, Schildberg FA, Odenthal M, Dienes HP, van Rooijen N, Schmitt E, Garbi N, Croft M, Kurts C, Kubes P, Protzer U, Heikenwalder M, Knolle PA. 2013. Intrahepatic myeloid cell-aggregates enable local CD8+ T cell expansion and successful immunotherapy against chronic viral liver infection. Nat Immunol. 14: 574-83.
Buchmann P, Dembek C, Kuklick L, Jäger C, Tedjokusumo R, von Freyend MJ, Drebber U, Janowicz Z, Melber K, Protzer U. 2013. A novel therapeutic hepatitis B vaccine induces cellular and humoral immune responses and breaks tolerance in HBV transgenic mice. Vaccine. 31:1197-1203.
Huang LR, Gäbel YA, Graf S, Arzberger S, Kurts C, Heikenwalder M, Knolle PA, Protzer U. 2012. Trans- fer of HBV genomes using low doses of adenovirus vectors leads to persistent infection in immune com- petent mice. Gastroenterology 142:1447-50.
von Freyend MJ, Untergasser A, Arzberger S, Oberwinkler H, Drebber U, Schirmacher P, Protzer U. 2011. Sequential control of hepatitis B virus in a mouse model of acute, self-resolving hepatitis B. J Viral Hepat 18:216-26.
Grimm D, Lee JS, Wang L, Desai T, Akache B, Storm TA, Kay MA. 2008. In vitro and in vivo gene thera- py vector evolution via multispecies interbreeding and re-targeting of adeno-associated viruses. J Virol 82:5887-911.
Grimm D, Streetz KL, Jopling CL, Storm TA, Pandey K, Davis CR, Marion P, Salazar F, Kay MA. 2006. Fatality in mice due to oversaturation of cellular microRNA/short hairpin RNA pathways. Nature. 441:537- 41.