Ultrasound use increases the delivery and efficacy of GLA-encoding modified RNA in the treatment of Fabry disease

Abstract

Fabry disease is an X-linked recessive lysosomal storage disorder due to the deficiency of the enzyme alpha galactosidase A (a-GAL A). Consequently, the accumulation of its substrates globotriaosylceramide (Gb3) and lyso-globotriaosylceramide (lyso-Gb3) leads to multisystemic disease manifestations. The current standard of therapy is biweekly enzyme replacement of recombinant a-GAL A, which may result in the development of antibodies, adverse injection reactions, economic burden, and patient inconvenience. The development of modified RNA-based therapy improved the delivery of mRNA and posed as a promising treatment method. This study aimed to investigate the organ-specific delivery of microbubble-coated mRNA to the heart using ultrasound probe to burst the bubbles, known as sonoporation. iPSCs from two Fabry patients were differentiated into diseased cardiomyocytes (iPSCCM) and transfected with modRNA to prove the efficacy of GLA-modRNA in vitro with the restoration of a-GAL A enzyme level compared to that of wildtype. The pharmacokinetic study demonstrated that ultrasound-assisted sonoporation of SonoVueTM microbubble-coated modRNA resulted in superior cardiac delivery of mRNA compared to intravenous administration or without microbubbles carriers. The in vivo efficacy of microbubbles-encapsulated modRNA was further demonstrated in transgenic GLA-KO mice model with higher enzyme level, demonstratable reduction in substrate level, and lack of immunogenicity compared to wildtype and enzyme replacement therapy. These results proved modRNA-based therapy to be an effective treatment method for Fabry patients. Further studies should focus on the development of animal models with clinical phenotypes resembling that of Fabry patients to measure the correlation between biochemical response and clinical improvement.