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Expression Of A Secretable, Cell-Penetrating Cdkl5 Protein Enhances The Efficacy Of Aav Vector-Mediated Gene Therapy For Cdkl5 Deficiency Disorder.

Medici, G.; Tassinari, M.; Galvani, G.; Bastianini, S.; Gennaccaro, L.; Loi, M.; Fuchs, C.; Mottolese, N.; Alvente, S.; Berteotti, C.; Bagget, H. R.; Zoccoli, G.; Giustetto, M.; Muotri, A. R.; Nakai, H.; Trazzi, S.; Ciani, E.

2021-07-26 neuroscience
10.1101/2021.07.26.453746 bioRxiv
Show abstract

No therapy is currently available for CDKL5 (cyclin-dependent kinase-like 5) deficiency disorder (CDD), a severe neurodevelopmental disorder caused by mutations in the CDKL5 gene. Although delivery of a wild-type copy of the mutated gene to cells represents the most curative approach for a monogenic disease, proof-of-concept studies highlight significant efficacy caveats for brain gene therapy. Herein, we used a secretable TATk-CDKL5 protein to enhance the efficiency of a gene therapy for CDD. We found that, although AAVPHP.B_Igk-TATk-CDKL5 and AAVPHP.B_CDKL5 vectors had similar brain infection efficiency, the AAVPHP.B_Igk-TATk-CDKL5 vector led to a higher CDKL5 protein replacement due to secretion and transduction of the TATk-CDKL5 protein into the neighboring cells. Importantly, Cdkl5 KO mice treated with the AAVPHP.B_Igk-TATk-CDKL5 vector showed a behavioral and neuroanatomical improvement in comparison with vehicle-treated Cdkl5 KO mice or Cdkl5 KO mice treated with the AAVPHP.B_CDKL5 vector, indicating that a gene therapy based on a secretable recombinant TATk-CDKL5 protein is more effective at compensating Cdkl5-null brain defects than gene therapy based on the expression of the native CDKL5.

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