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AAV-vectored expression of monospecific or bispecific monoclonal antibodies protects mice from lethal Pseudomonas aeruginosa pneumonia

Gene Therapy (2024)Cite this article

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Abstract

Pseudomonas aeruginosa poses a significant threat to immunocompromised individuals and those with cystic fibrosis. Treatment relies on antibiotics, but persistent infections occur due to intrinsic and acquired resistance of P. aeruginosa towards multiple classes of antibiotics. To date, there are no licensed vaccines for this pathogen, prompting the urgent need for novel treatment approaches to combat P. aeruginosa infection and persistence. Here we validated AAV vectored immunoprophylaxis as a strategy to generate long-term plasma and mucosal expression of highly protective monoclonal antibodies (mAbs) targeting the exopolysaccharide Psl (Cam-003) and the PcrV (V2L2MD) component of the type-III secretion system injectosome either as single mAbs or together as a bispecific mAb (MEDI3902) in a mouse model. When administered intramuscularly, AAV-αPcrV, AAV-αPsl, and AAV-MEDI3902 significantly protected mice challenged intranasally with a lethal dose of P. aeruginosa strains PAO1 and PA14 and reduced bacterial burden and dissemination to other organs. While all AAV-mAbs provided protection, AAV-αPcrV and AAV-MEDI3902 provided 100% and 87.5% protection from a lethal challenge with 4.47 × 107 CFU PAO1 and 87.5% and 75% protection from a lethal challenge with 3 × 107 CFU PA14, respectively. Serum concentrations of MEDI3902 were ~10× lower than that of αPcrV, but mice treated with this vector showed a greater reduction in bacterial dissemination to the liver, lung, spleen, and blood compared to other AAV-mAbs. These results support further investigation into the use of AAV vectored immunoprophylaxis to prevent and treat P. aeruginosa infections and other bacterial pathogens of public health concern for which current treatment strategies are limited.

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Fig. 1: AAV-αPcrV and AAV-αPsl mAb expression kinetics in the blood and bronchoalveolar lavage fluid (BALF) of transduced mice and confirmation of functional binding to P. aeruginosa.
Fig. 2: In vitro analysis of AAV6.2FF vectors expressing AAV-GFP-WPRE, AAV-GFP-WPRE3 or AAV-GFP (without a WPRE).
Fig. 3: Kinetic analysis and functional binding of AAV-expressed P. aeruginosa bispecific mAb, MEDI3902.
Fig. 4: AAV6.2FF-mAbs confer significant protection in two lethal P. aeruginosa pneumonia challenge models.
Fig. 5: AAV-αPcrV + AAV-αPsl and AAV-MEDI3902 provide excellent protection in the prevention of bacterial burden in the lungs and dissemination to other organs, while AAV-αPcrV and AAV-αPsl provide moderate protection.

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All data generated or analyzed during this study can be found within the published article and its supplementary files.

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Acknowledgements

Funding was provided by the Natural Sciences and Engineering Research Council of Canada (NSERC) Discover Grant (grant #499834), the Canadian Institutes of Health Research (CIHR) (grant # 202109PJ4-474300-VVP-CEHA-140973 to SKW and PJT 156111 to CMK), Mason Research Fund (grant #44610), and Cystic Fibrosis Canada (grant #1145528). Stipend support was provided by the Ontario Veterinary College (OVC) (JAL, JGEY, ESBC, ADR, BAYS), the Ontario Graduate Scholarship program (JGEY, MMG), the NSERC Postgraduate Scholarship-Doctoral program (JGEY, BAYS). We thank the technicians at the University of Guelph Animal Isolation Unit for their animal care services.

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Authors and Affiliations

  1. Department of Pathobiology, University of Guelph, Guelph, ON, N1G 2W1, Canada

    Jordyn A. Lopes, Yanlong Pei, Jacob G. E. Yates, Elena S. B. Campbell, Melanie M. Goens, Madison E. Hughes, Amira D. Rghei, Brenna A. Y. Stevens, Matthew M. Guilleman, Leonardo Susta & Sarah K. Wootton

  2. Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, N1G 2W1, Canada

    Nicole E. Garnier & Cezar M. Khursigara

  3. Avamab Pharma Inc., 120, 4838 Richard Road SW, Calgary, AB, T3E 6L1, Canada

    Brad Thompson

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Contributions

Conceptualization, JAL and SKW; methodology, JAL produced the vector and executed the study designs; NEG provided all bacterial samples and completed CFU enumeration for biodistribution study; JAL and JGEY ran flow cytometric analysis; JAL, NEG YP, JGEY, ESBC, MMG, MEH, ADR and BAYS assisted with vector cloning, animal work and/or data analysis. Writing-original draft preparation, JAL; writing-review and editing, MMG, BT, CMK, LS, and SKW; supervision, CMK, and SKW; funding acquisition, SKW. All authors have read and agreed to the published version of the manuscript.

Corresponding author

Correspondence to Sarah K. Wootton.

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Competing interests

SKW is an inventor on a US patent for the AAV6.2FF capsid. This patent (US20190216949) is licensed to Avamab Pharma Inc., where BT and SKW are co-founders and BT serves as an executive. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Ethical approval

This study was conducted according to the guidelines set forth by the Canadian Council on Animal Care (CCAC) and approved by the Animal Care Committee of the University of Guelph (Animal Use Protocol number 4664).

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Lopes, J.A., Garnier, N.E., Pei, Y. et al. AAV-vectored expression of monospecific or bispecific monoclonal antibodies protects mice from lethal Pseudomonas aeruginosa pneumonia. Gene Ther (2024). https://doi.org/10.1038/s41434-024-00453-1

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