Adenoviruses using the cancer marker EphA2 as a receptor in vitro and in vivo by genetic ligand insertion into different capsid scaffolds.
Adenoviral gene therapy and oncolysis would critically benefit from targeted cell entry by genetically modified capsids. This requires both the ablation of native adenovirus tropism and the identification of ligands that remain functional in virus context. Here, we establish cell type-specific entry of HAdV-5-based vectors by genetic ligand insertion into a chimeric fiber with shaft and knob domains of the short HAdV-41 fiber (Ad5T/41sSK). This fiber format was reported to ablate transduction in vitro and biodistribution to the liver in vivo. We show that the YSA peptide, binding to the pan-cancer marker EphA2, can be inserted into three positions of the chimeric fiber, resulting in strong transduction of EphA2-positive but not EphA2-negative cells of human melanoma biopsies and of tumor xenografts after intratumoral injection. Transduction was blocked by soluble YSA peptide and restored for EphA2-negative cells after recombinant EphA2 expression. The YSA peptide could also be inserted into three positions of a CAR binding-ablated HAdV-5 fiber enabling specific transduction; however, the Ad5T/41sSK format was superior in vivo. In conclusion, we establish an adenovirus capsid facilitating functional insertion of targeting peptides and a novel adenovirus using the tumor marker EphA2 as receptor with high potential for cancer gene therapy and viral oncolysis.
Oncolytic Adenovirus Group, German Cancer Research Center (DKFZ), Heidelberg, Germany.
Michael
M
Behr
Oncolytic Adenovirus Group, German Cancer Research Center (DKFZ), Heidelberg, Germany.
Johanna K
JK
Kaufmann
Oncolytic Adenovirus Group, German Cancer Research Center (DKFZ), Heidelberg, Germany.
Patrick
P
Ketzer
Oncolytic Adenovirus Group, German Cancer Research Center (DKFZ), Heidelberg, Germany.
Sarah
S
Engelhardt
Max von Pettenkofer-Institute, Department of Virology, Ludwig-Maximilians-University Munich, Munich, Germany.
Martin
M
Mück-Häusl
Department of General Pediatrics, Division of Inherited Metabolic Diseases, Heidelberg University Hospital, Heidelberg, Germany.
Pamela M
PM
Okun
COS, CellNetworks Deep Sequencing Core Facility, University Heidelberg, Heidelberg, Germany.
Gabriele
G
Petersen
Institute for Clinical and Molecular Virology, Erlangen University Hospital, Erlangen, Germany.
Frank
F
Neipel
Department of Dermatology, Heidelberg University Hospital, Heidelberg, Germany.
Jessica C
JC
Hassel
Max von Pettenkofer-Institute, Department of Virology, Ludwig-Maximilians-University Munich, Munich, Germany; Institute of Virology and Microbiology, Center for Biomedical Education and Research, Department of Human Medicine, Faculty of Health, University Witten/Herdecke, Witten, Germany.
Anja
A
Ehrhardt
Department of Dermatology, Heidelberg University Hospital, Heidelberg, Germany.
Alexander H
AH
Enk
Oncolytic Adenovirus Group, German Cancer Research Center (DKFZ), Heidelberg, Germany.
Dirk M
DM
Nettelbeck
D000256:Adenoviridae / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D005260:Female; D006801:Humans; D008545:Melanoma / Q000378:metabolism / Q000628:therapy; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D036104:Receptor, EphA2 / Q000378:metabolism*; D023041:Xenograft Model Antitumor Assays
D036104:Receptor, EphA2