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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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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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.
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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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DOI: https://doi.org/10.1038/s41434-021-00286-2
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