Cell-penetrating peptide-conjugated, splice-switching oligonucleotides mitigate the phenotype in BTK/Tec double deficient X-linked agammaglobulinemia model.

Bestas, BurcuBurcuBestasEstupiñán, H YesidH YesidEstupiñánWang, QingQingWangKharazi, ShabnamShabnamKharaziHe, ChenfeiChenfeiHeK Mohammad, DaraDaraK MohammadGupta, DhanuDhanuGuptaWiklander, Oscar P BOscar P BWiklanderLehto, TaaviTaaviLehtoLundin, Karin EKarin ELundinBerglöf, AnnaAnnaBerglöfKarlsson, Mikael C IMikael C IKarlssonAbendroth, FrankFrankAbendrothEl Andaloussi, SamirSamirEl AndaloussiGait, Michael JMichael JGaitWood, Matthew J AMatthew J AWoodLeumann, ChristianChristianLeumann0000-0002-7996-7083Stetsenko, Dmitry ADmitry AStetsenkoMånsson, RobertRobertMånssonWengel, JesperJesperWengelZain, RulaRulaZainSmith, C I EdvardC I EdvardSmith2025-05-012025-05-012025-05-08https://boris-portal.unibe.ch/handle/20.500.12422/209494Splice-switching oligonucleotides (SSOs) have been developed as a treatment for various disorders, including Duchenne muscular dystrophy and spinal muscular atrophy. Here, the activity of several different SSOs was investigated as potential treatments for B lymphocyte disorders with a focus on X-linked agammaglobulinemia (XLA), caused by defects in the gene encoding Bruton's tyrosine kinase (BTK). In this study, the activity of locked nucleic acid (LNA), tricyclo-DNA (tcDNA), phosphoryl guanidine oligonucleotides (PGO) and phosphorodiamidate morpholino oligomers (PMO) were compared, targeting the pseudoexon region of BTK pre-mRNA. We further investigated the effect of conjugating cell-penetrating peptides, including Pip6a, to the SSOs. The effect was measured as splice-switching in vitro as well as in a further developed, bacterial artificial chromosome transgenic mouse model of XLA. Therapy in the form of intravenous infusions 2 times a week during 3 weeks of PMO oligomers conjugated to Pip6a was sufficient to partly restore the in vivo B lineage phenotype. SSOs treatment also provides a unique opportunity to get insights into a restoration process, when B lymphocytes of different maturation stages are simultaneously splice-corrected.en500 - Science::540 - Chemistry500 - Science::570 - Life sciences; biologyCell-penetrating peptide-conjugated, splice-switching oligonucleotides mitigate the phenotype in BTK/Tec double deficient X-linked agammaglobulinemia model.article10.48620/877254017124810.1039/d4cb00312h