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Virotherapy research involves the development, exploration, and application of oncolytic viruses that combine direct killing of cancer cells by viral infection, replication, and spread (oncolysis) with indirect killing by induction of anti-tumor immune responses. Oncolytic viruses can also be engineered to genetically deliver therapeutic proteins for direct or indirect cancer cell killing. In this review—as part of the special edition on “State-of-the-Art Viral Vector Gene Therapy in Germany”—the German community of virotherapists provides an overview of their recent research activities that cover endeavors from screening and engineering viruses as oncolytic cancer therapeutics to their clinical translation in investigator-initiated and sponsored multi-center trials. Preclinical research explores multiple viral platforms, including new isolates, serotypes, or fitness mutants, and pursues unique approaches to engineer them towards increased safety, shielded or targeted delivery, selective or enhanced replication, improved immune activation, delivery of therapeutic proteins or RNA, and redirecting antiviral immunity for cancer cell killing. Moreover, several oncolytic virus-based combination therapies are under investigation. Clinical trials in Germany explore the safety and potency of virotherapeutics based on parvo-, vaccinia, herpes, measles, reo-, adeno-, vesicular stomatitis, and coxsackie viruses, including viruses encoding therapeutic proteins or combinations with immune checkpoint inhibitors. These research advances represent exciting vantage points for future endeavors of the German virotherapy community collectively aimed at the implementation of effective virotherapeutics in clinical oncology.
Dirk Nettelbeck; Mathias Leber; Jennifer Altomonte; Assia Angelova; Julia Beil; Susanne Berchtold; Maike Delic; Jürgen Eberle; Anja Ehrhardt; Christine Engeland; Henry Fechner; Karsten Geletneky; Katrin Goepfert; Per Holm; Stefan Kochanek; Florian Kreppel; Lea Krutzke; Florian Kühnel; Karl Lang; Antonio Marchini; Markus Moehler; Michael Mühlebach; Ulrike Naumann; Roman Nawroth; Jürg Nüesch; Jean Rommelaere; Ulrich Lauer; Guy Ungerechts. Virotherapy in Germany—Recent Activities in Virus Engineering, Preclinical Development, and Clinical Studies. Viruses 2021, 13, 1420 .
AMA StyleDirk Nettelbeck, Mathias Leber, Jennifer Altomonte, Assia Angelova, Julia Beil, Susanne Berchtold, Maike Delic, Jürgen Eberle, Anja Ehrhardt, Christine Engeland, Henry Fechner, Karsten Geletneky, Katrin Goepfert, Per Holm, Stefan Kochanek, Florian Kreppel, Lea Krutzke, Florian Kühnel, Karl Lang, Antonio Marchini, Markus Moehler, Michael Mühlebach, Ulrike Naumann, Roman Nawroth, Jürg Nüesch, Jean Rommelaere, Ulrich Lauer, Guy Ungerechts. Virotherapy in Germany—Recent Activities in Virus Engineering, Preclinical Development, and Clinical Studies. Viruses. 2021; 13 (8):1420.
Chicago/Turabian StyleDirk Nettelbeck; Mathias Leber; Jennifer Altomonte; Assia Angelova; Julia Beil; Susanne Berchtold; Maike Delic; Jürgen Eberle; Anja Ehrhardt; Christine Engeland; Henry Fechner; Karsten Geletneky; Katrin Goepfert; Per Holm; Stefan Kochanek; Florian Kreppel; Lea Krutzke; Florian Kühnel; Karl Lang; Antonio Marchini; Markus Moehler; Michael Mühlebach; Ulrike Naumann; Roman Nawroth; Jürg Nüesch; Jean Rommelaere; Ulrich Lauer; Guy Ungerechts. 2021. "Virotherapy in Germany—Recent Activities in Virus Engineering, Preclinical Development, and Clinical Studies." Viruses 13, no. 8: 1420.
Adenovirus-based vectors are playing an important role as efficacious genetic vaccines to fight the current COVID-19 pandemic. Furthermore, they have an enormous potential as oncolytic vectors for virotherapy and as vectors for classic gene therapy. However, numerous vector–host interactions on a cellular and noncellular level, including specific components of the immune system, must be modulated in order to generate safe and efficacious vectors for virotherapy or classic gene therapy. Importantly, the current widespread use of Ad vectors as vaccines against COVID-19 will induce antivector immunity in many humans. This requires the development of strategies and techniques to enable Ad-based vectors to evade pre-existing immunity. In this review article, we discuss the current status of genetic and chemical capsid modifications as means to modulate the vector–host interactions of Ad-based vectors.
Denice Weklak; Daniel Pembaur; Georgia Koukou; Franziska Jönsson; Claudia Hagedorn; Florian Kreppel. Genetic and Chemical Capsid Modifications of Adenovirus Vectors to Modulate Vector–Host Interactions. Viruses 2021, 13, 1300 .
AMA StyleDenice Weklak, Daniel Pembaur, Georgia Koukou, Franziska Jönsson, Claudia Hagedorn, Florian Kreppel. Genetic and Chemical Capsid Modifications of Adenovirus Vectors to Modulate Vector–Host Interactions. Viruses. 2021; 13 (7):1300.
Chicago/Turabian StyleDenice Weklak; Daniel Pembaur; Georgia Koukou; Franziska Jönsson; Claudia Hagedorn; Florian Kreppel. 2021. "Genetic and Chemical Capsid Modifications of Adenovirus Vectors to Modulate Vector–Host Interactions." Viruses 13, no. 7: 1300.
Adenovirus-based gene transfer vectors are the most frequently used vector type in gene therapy clinical trials to date, and they play an important role as genetic vaccine candidates during the ongoing SARS-CoV-2 pandemic. Immediately upon delivery, adenovirus-based vectors exhibit multiple complex vector-host interactions and induce innate and adaptive immune responses. This can severely limit their safety and efficacy, particularly after delivery through the blood stream. In this review article we summarize two strategies to modulate Ad vector-induced immune responses: extensive genomic and chemical capsid modifications. Both strategies have shown beneficial effects in a number of preclinical studies while potential synergistic effects warrant further investigations.
Florian Kreppel; Claudia Hagedorn. Capsid and Genome Modification Strategies to Reduce the Immunogenicity of Adenoviral Vectors. International Journal of Molecular Sciences 2021, 22, 2417 .
AMA StyleFlorian Kreppel, Claudia Hagedorn. Capsid and Genome Modification Strategies to Reduce the Immunogenicity of Adenoviral Vectors. International Journal of Molecular Sciences. 2021; 22 (5):2417.
Chicago/Turabian StyleFlorian Kreppel; Claudia Hagedorn. 2021. "Capsid and Genome Modification Strategies to Reduce the Immunogenicity of Adenoviral Vectors." International Journal of Molecular Sciences 22, no. 5: 2417.
The surface of proteins is heterogeneous with sophisticated but precise hydrophobic and hydrophilic patches, which is essential for their diverse biological functions. To emulate such distinct surface patterns on macromolecules, we used rigid spherical synthetic dendrimers (polyphenylene dendrimers) to provide controlled amphiphilic surface patches with molecular precision. We identified an optimal spatial arrangement of these patches on certain dendrimers that enabled their interaction with human adenovirus 5 (Ad5). Patchy dendrimers bound to the surface of Ad5 formed a synthetic polymer corona that greatly altered various host interactions of Ad5 as well as in vivo distribution. The dendrimer corona (1) improved the ability of Ad5-derived gene transfer vectors to transduce cells deficient for the primary Ad5 cell membrane receptor and (2) modulated the binding of Ad5 to blood coagulation factor X, one of the most critical virus-host interactions in the bloodstream. It significantly enhanced the transduction efficiency of Ad5 while also protecting it from neutralization by natural antibodies and the complement system in human whole blood. Ad5 with a synthetic dendrimer corona revealed profoundly altered in vivo distribution, improved transduction of heart, and dampened vector sequestration by liver and spleen. We propose the design of bioactive polymers that bind protein surfaces solely based on their amphiphilic surface patches and protect against a naturally occurring protein corona, which is highly attractive to improve Ad5-based in vivo gene therapy applications.
Yuzhou Wu; Longjie Li; Larissa Frank; Jessica Wagner; Patrizia Andreozzi; Brenton A. G. Hammer; Marco D’Alicarnasso; Maria Pelliccia; Weina Liu; Sabyasachi Chakrabortty; Silke Krol; Johanna Simon; Katharina Landfester; Seah Ling Kuan; Francesco Stellacci; Klaus Müllen; Florian Kreppel; Tanja Weil. Patchy Amphiphilic Dendrimers Bind Adenovirus and Control Its Host Interactions and in Vivo Distribution. ACS Nano 2019, 13, 8749 -8759.
AMA StyleYuzhou Wu, Longjie Li, Larissa Frank, Jessica Wagner, Patrizia Andreozzi, Brenton A. G. Hammer, Marco D’Alicarnasso, Maria Pelliccia, Weina Liu, Sabyasachi Chakrabortty, Silke Krol, Johanna Simon, Katharina Landfester, Seah Ling Kuan, Francesco Stellacci, Klaus Müllen, Florian Kreppel, Tanja Weil. Patchy Amphiphilic Dendrimers Bind Adenovirus and Control Its Host Interactions and in Vivo Distribution. ACS Nano. 2019; 13 (8):8749-8759.
Chicago/Turabian StyleYuzhou Wu; Longjie Li; Larissa Frank; Jessica Wagner; Patrizia Andreozzi; Brenton A. G. Hammer; Marco D’Alicarnasso; Maria Pelliccia; Weina Liu; Sabyasachi Chakrabortty; Silke Krol; Johanna Simon; Katharina Landfester; Seah Ling Kuan; Francesco Stellacci; Klaus Müllen; Florian Kreppel; Tanja Weil. 2019. "Patchy Amphiphilic Dendrimers Bind Adenovirus and Control Its Host Interactions and in Vivo Distribution." ACS Nano 13, no. 8: 8749-8759.
Adenovirus vectors are potent tools for genetic vaccination and oncolytic virotherapy. However, they are prone to multiple undesired vector-host interactions, especially after in vivo delivery. It is a consensus that the limitations imposed by undesired vector-host interactions can only be overcome if defined modifications of the vector surface are performed. These modifications include shielding of the particles from unwanted interactions and targeting by the introduction of new ligands. The goal of the protocol presented here is to enable the reader to generate shielded and, if desired, retargeted human adenovirus gene transfer vectors or oncolytic viruses. The protocol will enable researchers to modify the surface of adenovirus vector capsids by specific chemical attachment of synthetic polymers, carbohydrates, lipids, or other biological or chemical moieties. It describes the cutting-edge technology of combined genetic and chemical capsid modifications, which have been shown to facilitate the understanding and overcoming of barriers for in vivo delivery of adenovirus vectors. A detailed and commented description of the crucial steps for performing specific chemical reactions with biologically active viruses or virus-derived vectors is provided. The technology described in the protocol is based on the genetic introduction of (naturally absent) cysteine residues into solvent-exposed loops of adenovirus-derived vectors. These cysteine residues provide a specific chemical reactivity that can, after production of the vectors to high titers, be exploited for highly specific and efficient covalent chemical coupling of molecules from a wide variety of substance classes to the vector particles. Importantly, this protocol can easily be adapted to perform a broad variety of different (non-thiol-based) chemical modifications of adenovirus vector capsids. Finally, it is likely that non-enveloped virus-based gene transfer vectors other than adenovirus can be modified from the basis of this protocol.
Franziska Jönsson; Claudia Hagedorn; Florian Kreppel. Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting. Journal of Visualized Experiments 2018, e58480 -e58480.
AMA StyleFranziska Jönsson, Claudia Hagedorn, Florian Kreppel. Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting. Journal of Visualized Experiments. 2018; (140):e58480-e58480.
Chicago/Turabian StyleFranziska Jönsson; Claudia Hagedorn; Florian Kreppel. 2018. "Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting." Journal of Visualized Experiments , no. 140: e58480-e58480.
Adenovirus-based vectors comprise the most frequently used vector type in clinical studies to date. Both intense lab research and insights from the clinical trials reveal the importance of a comprehensive understanding of vector–host interactions. Especially for systemic intravenous adenovirus vector delivery, it is paramount to develop safe and efficacious vectors. Very early vector–host interactions that take place in blood long before the first cell is being transduced are phenomena triggered by the surface, shape, and size of the adenovirus vector particles. Not surprisingly, a multitude of different technologies ranging from genetics to chemistry has been developed to alter the adenovirus vector surface. In this review, we discuss the most important technologies and evaluate them for their suitability to overcome hurdles imposed by early vector–host interactions.
Claudia Hagedorn; Florian Kreppel. Capsid Engineering of Adenovirus Vectors: Overcoming Early Vector–Host Interactions for Therapy. Human Gene Therapy 2017, 28, 820 -832.
AMA StyleClaudia Hagedorn, Florian Kreppel. Capsid Engineering of Adenovirus Vectors: Overcoming Early Vector–Host Interactions for Therapy. Human Gene Therapy. 2017; 28 (10):820-832.
Chicago/Turabian StyleClaudia Hagedorn; Florian Kreppel. 2017. "Capsid Engineering of Adenovirus Vectors: Overcoming Early Vector–Host Interactions for Therapy." Human Gene Therapy 28, no. 10: 820-832.
Vectors based on human adenovirus are highly efficient tools for transient genetic modifications of cells or tissues in vitro and in vivo. They can be utilized for gene addition strategies, knockdown strategies and as transfer vectors for designer nucleases and CRISPR/Cas. They are characterized by high genomic stability and can be produced to high titers. This chapter describes the method how to produce, purify and titrate adenovirus vectors based on human adenovirus type 5.
Ramona F. Kratzer; Florian Kreppel. Production, Purification, and Titration of First-Generation Adenovirus Vectors. Methods in Molecular Biology 2017, 1654, 377 -388.
AMA StyleRamona F. Kratzer, Florian Kreppel. Production, Purification, and Titration of First-Generation Adenovirus Vectors. Methods in Molecular Biology. 2017; 1654 ():377-388.
Chicago/Turabian StyleRamona F. Kratzer; Florian Kreppel. 2017. "Production, Purification, and Titration of First-Generation Adenovirus Vectors." Methods in Molecular Biology 1654, no. : 377-388.
Florian Kreppel; Anja Ehrhardt. From Virus to vector to medicine: Foreword by guest editors. Virus Genes 2017, 53, 673 -674.
AMA StyleFlorian Kreppel, Anja Ehrhardt. From Virus to vector to medicine: Foreword by guest editors. Virus Genes. 2017; 53 (5):673-674.
Chicago/Turabian StyleFlorian Kreppel; Anja Ehrhardt. 2017. "From Virus to vector to medicine: Foreword by guest editors." Virus Genes 53, no. 5: 673-674.
Currently, virus-based vectors, namely derivatives of the adenovirus, are frequently used in a wide variety of ex vivo or local gene therapeutic applications. However, the efficacy of virus-based vectors in systemic applications is presently still extremely limited. Complex interactions of the various vector types with the patient’s organism hinder successful vector deployment. Exemplary, here we summarize barriers to systemic application of Adenovirus-based vectors leading either to acute toxic effects or rapid vector neutralization and discuss strategies to overcome these barriers aiming to develop more efficient vector types.
Franziska Jönsson; Florian Kreppel. Barriers to systemic application of virus-based vectors in gene therapy: lessons from adenovirus type 5. Virus Genes 2017, 53, 692 -699.
AMA StyleFranziska Jönsson, Florian Kreppel. Barriers to systemic application of virus-based vectors in gene therapy: lessons from adenovirus type 5. Virus Genes. 2017; 53 (5):692-699.
Chicago/Turabian StyleFranziska Jönsson; Florian Kreppel. 2017. "Barriers to systemic application of virus-based vectors in gene therapy: lessons from adenovirus type 5." Virus Genes 53, no. 5: 692-699.
Adenovirus-based vectors are promising tools for genetic vaccination. However, several obstacles have to be overcome prior to a routine clinical application of adenovirus-based vectors as efficacious vectored vaccines. The linear trisaccharide epitope αGal (alpha-Gal) with the carbohydrate sequence galactose-α-1,3-galactosyl-β-1,4-N-acetylglucosamine has been described as a potent adjuvant for recombinant or attenuated vaccines. Humans and α-1,3-galactosyltransferase knockout mice do not express this epitope. Upon exposure of α-1,3-galactosyltransferase-deficient organisms to αGal in the environment, large amounts of circulating anti-Gal antibodies are produced consistently. Immunocomplexes formed between recombinant αGal-decorated vaccines and anti-Gal antibodies exhibit superior immunogenicity. We studied the effects of the trisaccharide epitope on CD8 T cell responses that are directed specifically to vector-encoded transgenic antigens. For that, covalently αGal-decorated adenovirus vectors were delivered to anti-Gal α-1,3-galactosyltransferase knockout mice. We generated replication-defective, E1-deleted adenovirus type 5 vectors that were decorated with αGal at the hexon hypervariable regions 1 or 5, at fiber knob, or at penton base. Surprisingly, none of the adenovirus immunocomplexes being formed from αGal-decorated adenovirus vectors and anti-Gal immunoglobulins improved the frequencies of CD8 T cell responses against the transgenic antigen ovalbumin. Humoral immunity directed to the adenovirus vector was neither increased. However, our data indicated that decoration of Ad vectors with the αGal epitope is a powerful tool to analyze the fate of adenovirus immunocomplexes in vivo.
Ramona F. Kratzer; Sigrid Espenlaub; Andrea Hoffmeister; Matthias W. Kron; Florian Kreppel. Covalent decoration of adenovirus vector capsids with the carbohydrate epitope αGal does not improve vector immunogenicity, but allows to study the in vivo fate of adenovirus immunocomplexes. PLOS ONE 2017, 12, e0176852 .
AMA StyleRamona F. Kratzer, Sigrid Espenlaub, Andrea Hoffmeister, Matthias W. Kron, Florian Kreppel. Covalent decoration of adenovirus vector capsids with the carbohydrate epitope αGal does not improve vector immunogenicity, but allows to study the in vivo fate of adenovirus immunocomplexes. PLOS ONE. 2017; 12 (5):e0176852.
Chicago/Turabian StyleRamona F. Kratzer; Sigrid Espenlaub; Andrea Hoffmeister; Matthias W. Kron; Florian Kreppel. 2017. "Covalent decoration of adenovirus vector capsids with the carbohydrate epitope αGal does not improve vector immunogenicity, but allows to study the in vivo fate of adenovirus immunocomplexes." PLOS ONE 12, no. 5: e0176852.