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Correction of a knock-in mouse model of acrodysostosis with gene therapy using a rAAV9-CAG-human PRKAR1A vector

Gene Therapy volume 29, pages 441–448 (2022)Cite this article

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Abstract

Acrodysostosis is a rare skeletal dysplasia caused by loss-of-function mutations in the regulatory subunit of protein kinase A (PRKAR1A). In a knock-in mouse model (PRKAR1Awt/mut) expressing one copy of the recurrent R368X mutation, we tested the effects of a rAAV9-CAG-human PRKR1A (hPRKAR1A) vector intravenously administered at 4 weeks of age. Caudal vertebrae and tibial diaphyses contained 0.52 ± 0.7 and 0.13 ± 0.3 vector genome per cell (VGC), respectively, at 10 weeks of age and 0.22 ± 0.04 and 0.020 ± 0.04 at 16 weeks while renal cortex contained 0.57 ± 0.14 and 0.26 ± 0.05 VGC. Vector-mediated hPRKAR1A expression was found in growth plate chondrocytes, osteoclasts, osteoblasts, and kidney tubular cells. Chondrocyte architecture was restored in the growth plates. Body length, tail length, and body weight were improved in vector treated PRKAR1Awt/mut mice, not the bone length of their limbs. These results provide one of the few proofs for gene therapy efficacy in a mouse model of chondrodysplasia. In addition, the increased urinary cAMP of PRKAR1Awt/mut mice was corrected almost to normal. In conclusion, gene therapy with hPRKAR1A improved skeletal growth and kidney dysfunction, the hallmarks of acrodysostosis in R368X mutated mice and humans.

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Fig. 1: Effects of gene therapy on growth plate histology and vector expression.
Fig. 2: Immunofluorescence images of renal cortex showing DAPI (blue nuclei) and anti-hPRKAR1A antibody (deep red) staining.
Fig. 3: Skeletal effects of gene therapy.
Fig. 4: Diminution of urinary cAMP in response to gene therapy.
Fig. 5: Diminution of PTH in response to gene therapy.

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Acknowledgements

We are pleased to acknowledge our former teammate C.Silve, who discovered the R368X mutation of PRKAR1A in patients with acrodysostosis and documented its functional effects, then conducted the first bone and renal studies in the mouse model of the disease. We thank Sophie Valtat for the figures. We thank P. Hantraye and R. Aaron-Badin for providing MIRCen platforms to our experimental studies, and F. Balter for vector production.

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Author notes

  1. These authors contributed equally: Yasemin Özgür-Günes, Catherine Le Stunff.

Authors and Affiliations

  1. UMR1195 Inserm and Université Paris Saclay, Le Kremlin-Bicêtre, France

    Yasemin Özgür-Günes, Catherine Le Stunff, Malha Chedik & Pierre Bougnères

  2. MIRCen Institute, CEA, Fontenay-aux-Roses, France

    Yasemin Özgür-Günes, Catherine Le Stunff & Pierre Bougnères

  3. Therapy Design Consulting, Vincennes, France

    Marie-Pierre Belot & Pierre Bougnères

  4. Biochemistry Laboratory, Hôpital Bicêtre, Assistance Publique-Hôpitaux de Paris, Le Kremlin-Bicêtre, France

    Pierre-Hadrien Becker

  5. Atlantic Gene Therapies, Nantes, France

    Véronique Blouin

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  1. Yasemin Özgür-Günes
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Contributions

PB, CLS, and YOG designed the experiments and wrote the paper. CLS and YOG built the CAG-hPRKAR1A plasmid. MC and YOG performed histochemistry and immunofuorescence studies. MPB measured VGC. CLS measured skeletal pieces, hormones, and urinary cAMP. PHB measured serum biochemistry. VB produced the AAV9 vector.

Corresponding author

Correspondence to Pierre Bougnères.

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Competing interests

PB discloses founding two companies (TherapyDesignConsulting in France, Adrenas Therapeutics in USA).

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Özgür-Günes, Y., Le Stunff, C., Chedik, M. et al. Correction of a knock-in mouse model of acrodysostosis with gene therapy using a rAAV9-CAG-human PRKAR1A vector. Gene Ther 29, 441–448 (2022). https://doi.org/10.1038/s41434-021-00286-2

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