ADEVO: Proof-of-concept of adenovirus-directed EVOlution by random peptide display on the fiber knob

Abstract

Directed evolution of viral vectors involves the generation of randomized libraries followed by artificial selection of improved variants. Directed evolution only yielded limited results in adenovirus vector (AdV) development until now, mainly due to insufficient complexities of randomized libraries. Clinical applications of AdVs as gene therapy or oncolytic vectors are still hampered by the predetermined tropism of natural types. To overcome this challenge, we hypothesized that the technology of randomized peptide insertions on the capsid surface can be incorporated into the AdV bioengineering toolbox for vector retargeting. Here we developed Adenovirus Directed EVOlution (ADEVO) protocols based on fiber knob peptide display. As a proof-of-concept, HAdV-C5-derived libraries were constructed following three distinct protocols and selected on A549-DCAR cells that lack the HAdV-C5 primary receptor, with the goal of identifying variants able to infect and lyse these tumor cells more efficiently. All protocols enabled the construction of high complexity libraries with up to 9.6x10^5 unique variants, an approximate 100-fold improvement compared to previously published AdV libraries. After selection, the most enriched variants did not display enhanced infectivity but rather more efficient replication and cell lysis. This warrants investigations into potential unsuspected involvement of the fiber protein in adenovirus replication. GRAPHICAL ABSTRACT

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BibTeX RIS

BibTeX

@article{sallard2026adevo,
  title = {{ADEVO: Proof-of-concept of adenovirus-directed EVOlution by random peptide display on the fiber knob}},
  author = {Erwan Sallard and Julian Fischer and Katrin Schroeer and Lisa-Marie Dawson and Nissai Beaude and Arsalene Affes and Eric Ehrke-Schulz and Wenli Zhang and Adrian Westhaus and Marti Cabanes-Creus and Leszek Lisowski and Zsolt Ruszics and Anja Ehrhardt},
  journal = {Molecular Therapy: Oncology},
  year = {2026},
  month = sep,
  doi = {10.1016/j.omton.2024.200867},
  url = {https://hal.science/hal-05772048v1},
  note = {CC BY},
}

RIS

TY  - JOUR
TI  - ADEVO: Proof-of-concept of adenovirus-directed EVOlution by random peptide display on the fiber knob
AU  - Erwan Sallard
AU  - Julian Fischer
AU  - Katrin Schroeer
AU  - Lisa-Marie Dawson
AU  - Nissai Beaude
AU  - Arsalene Affes
AU  - Eric Ehrke-Schulz
AU  - Wenli Zhang
AU  - Adrian Westhaus
AU  - Marti Cabanes-Creus
AU  - Leszek Lisowski
AU  - Zsolt Ruszics
AU  - Anja Ehrhardt
PY  - 2026
DA  - 2026/09/30
JO  - Molecular Therapy: Oncology
DO  - 10.1016/j.omton.2024.200867
UR  - https://hal.science/hal-05772048v1
AB  - Directed evolution of viral vectors involves the generation of randomized libraries followed by artificial selection of improved variants. Directed evolution only yielded limited results in adenovirus vector (AdV) development until now, mainly due to insufficient complexities of randomized libraries. Clinical applications of AdVs as gene therapy or oncolytic vectors are still hampered by the predetermined tropism of natural types. To overcome this challenge, we hypothesized that the technology of randomized peptide insertions on the capsid surface can be incorporated into the AdV bioengineering toolbox for vector retargeting. Here we developed Adenovirus Directed EVOlution (ADEVO) protocols based on fiber knob peptide display. As a proof-of-concept, HAdV-C5-derived libraries were constructed following three distinct protocols and selected on A549-DCAR cells that lack the HAdV-C5 primary receptor, with the goal of identifying variants able to infect and lyse these tumor cells more efficiently. All protocols enabled the construction of high complexity libraries with up to 9.6x10^5 unique variants, an approximate 100-fold improvement compared to previously published AdV libraries. After selection, the most enriched variants did not display enhanced infectivity but rather more efficient replication and cell lysis. This warrants investigations into potential unsuspected involvement of the fiber protein in adenovirus replication. GRAPHICAL ABSTRACT
SN  - 2950-3299
ER  -

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