Intracellular Delivery of Short Interfering RNA in Rat Organ of Corti Using a Cell-penetrating Peptide PepFect6

Molecular Therapy: Nucleic Acids

Anders Fridberger¹  Stefan Jacob¹  Staffan Lindberg²  Ülo Langel²  Mats Ulfendahl³  Suvarna Dash-Wagh³ 

[1] Department of Clinical Science, Intervention, and Technology, Karolinska Institutet, Stockholm, Sweden;Department of Neurochemistry, Stockholm University, Stockholm, Sweden;Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden;

关键词: cell-penetrating peptide (CPP);    cochlea;    connexin;    FRAP;    gap junctions;    gene therapy;    inner ear;    nanoparticles;    siRNA;   

DOI  :  10.1038/mtna.2012.50

来源: DOAJ

【 摘 要 】

RNA interference (RNAi) using short interfering RNA (siRNA) is an attractive therapeutic approach for treatment of dominant-negative mutations. Some rare missense dominant-negative mutations lead to congenital-hearing impairments. A variety of viral vectors have been tested with variable efficacy for modulating gene expression in inner ear. However, there is concern regarding their safety for clinical use. Here, we report a novel cell-penetrating peptide (CPP)-based nonviral approach for delivering siRNA into inner ear tissue using organotypic cultures as model system. PepFect6 (PF6), a variant of stearyl-TP10, was specially designed for improved delivery of siRNA by facilitating endosomal release. We show that PF6 was internalized by all cells without inducing cytotoxicity in cochlear cultures. PF6/siRNA nanoparticles lead to knockdown of target genes, a housekeeping gene and supporting cell-specific connexin 26. Interestingly, application of PF6/connexin 26 siRNA exhibited knockdown of both connexin 26 and 30 mRNA and their absence led to impaired intercellular communication as demonstrated by reduced transfer of calcein among the PF6/connexin 26-siRNA–treated cells. Thus, we conclude that PF6 is an efficient nonviral vector for delivery of siRNA, which can be applied as a tool for the development of siRNA-based therapeutic applications for hearing impairments.

【 授权许可】

Unknown