Project Description

TP18: Combinatorial knock-down/knock-out strategies to reconstitute antiviral im-munity and eliminate persisting hepatitis B virus cccDNA

We aim to overcome the immune tolerance against HBV by controlling HBV antigen expression. This will be achieved by inhibiting HBV on the RNA and DNA level via RNAi and CRISPR, respectively. We expect to find that direct targeting of the HBV cccDNA and suppression of the viral transcripts, and hence of viral antigen expression, will restore HBV immunity either on its own or at least after therapeutic vaccination, and that restored B- and T-cell immunity will ultimately clear HBV in vivo. Specific aims include AAV vector fine-tuning in increasingly complex HBV infection models, targeting of checkpoints by RNAi, characterization of potential adverse effects and improving CRISPR safety.
A. Wisskirchen K*, Kah J*, Malo A, Asen T, Volz T, Allweiss L, Wettengel JM, Lütgehetmann M, Urban S, Bauer T, Dandri M*, Protzer U*. 2019. T-Cell Receptor Grafting allows Virological Control of Hepatitis B Virus Infection. J Clin Invest. 129(7):2932-2945.
B. Hoffmann M, 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.
C. Kosinska AD, Moeed A, Kallin N, Festag J, Su J, Steiger K, Michel ML, Protzer U, Knolle PA. 2019. Synergy of therapeutic heterologous prime-boost hepatitis B vaccination with CpG-application to im-prove immune control of persistent HBV infection. Sci Rep. 9(1):10808.
D. Bubeck F, Hoffmann M, Harteveld Z, Aschenbrenner S, Bietz A, Waldhauer M, Börner K, Fakhiri J, Schmelas C, Dietz L, Grimm D, Correia B, Eils R, Niopek D. 2018. Engineered anti-CRISPR proteins for optogenetic control of CRISPR/Cas9. Nat. Methods 15(11):924-927.
E. Schmelas C, Grimm D. 2018. Split Cas9, not hairs – advancing the therapeutic index of CRISPR tech-nology. Biotechnol. J. 13:e1700432.
F. Velkov S, Ott J, Protzer U, Michler T. 2018. The global Hepatitis B Virus genotype distribution estimat-ed from available genotyping data. Genes 9(10):pii: E495.
G. 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.
H. Burwitz B*, Wettengel J*, Mück-Häusl M*, Ringelhan M, Ko C, Festag MM, Hammond KB, Northrup M, Bimber BN, Jacob T, Reed JS, Reed N, Park B, Moller-Tank S, Esser K, Greene JM, Wu HL, Ab-dulhaqq S, Webb G, Sutton WF, Klug A, Swanson T, Legasse AW, Vu TQ, Asokan A, Haigwood NL, Protzer U*, Sacha J*. 2017. Hepatocytic Expression of Human Sodium-Taurocholate Cotransporting Polypeptide Enables Hepatitis B Virus Infection of Macaques. Nat. Commun. 8(1):2146.
I. Lempp FA, Wiedtke E, Qu B, Roques P, Chemin I, Vondran FWR, Grand RL, 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 66(3):703-716.
J. Michler T, Grosse S, Mockenhaupt S, Röder N, Stückler F, Knapp B, Ko C, Heikenwälder 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):1082-1098.