Structural optimization and additional targets identification of antisense oligonucleotide G3139 encapsulated in a neutral cytidinyl-lipid combined with a cationic lipid in vitro and in vivo. (March 2019)

• Record Type: Journal Article
• Authors: Ma, Yuan; Zhao, Wenting; Li, Yiding; Pan, Yufei; Wang, Shuhe; Zhu, Yuejie; Kong, Lingxuan; Guan, Zhu; Wang, Jiancheng; Zhang, Lihe; Yang, Zhenjun
• Is Part Of: Biomaterials. Volume 197(2019)
• Page Start: 182
• Page End: 193
• Publication Date: 2019-03
• Subjects: Gene delivery -- G3139 -- DNCA -- PS modification -- LNA-gapmer; Biomedical materials -- Periodicals; Biocompatible Materials -- Periodicals; Biomatériaux -- Périodiques; 610.28
• DOI: 10.1016/j.biomaterials.2018.12.033
• Languages: English
• ISSNs: 0142-9612

Abstract

Abstract: Antisense oligonucleotides (ASOs) usually contain a fully phosphorothioate (PS) backbone, which possibly interact with many genes and proteins under intracellular conditions. G3139 is an ASO that targets Bcl-2 mRNA and induces cell apoptosis. Here, we report a kind of cytidinyl-lipid combined with a cationic lipid (DNCA/CLD, molar ration, 28:3, named mix), which may interact with oligonucleotides via H-bond formation, pi-stacking and electrostatic interaction, accompanied by low zeta potentials. The IC50 value of G3139 delivered by mix-lipid reduced from above 20 μM to 0.158 μM for MCF-7/ADR, and exhibited stronger antiproliferation upon other cancer cell lines. In addition, PS modification in the 3′-half of G3139 (especially at positions 13-16) enhanced serum stability, target specificity and anticancer activity. Also, a locked nucleic acid (LNA) gapmer G3139 (LNA-G3139) showed superior antiproliferation (78.5%) and Bcl-2 mRNA suppression effects (85.5%) at 200 nM, mainly due to its high complementary RNA affinity. More apoptosis-associated targets were identified, and a lower level of non-specific protein binding (HSA) revealed that both antisense and aptamer mechanisms might simultaneously exist. A combination of a new delivery system and chemical modifications, such as in LNA-G3139, may have potential clinical application prospects in the future. Graphical abstract: Outlined description: Encapsulation and delivery by cytidinyl-lipid/cationic lipid (DNCA/CLD), ASOs phosphrothiolated in 3′-half region (such as site 13-16) significantly enhance target specificity and serum stability, while its LNA-gapmer (LNA-G3139) shows great mRNA affinity, inhibition of multiple targets expression (Bcl-2, ATAD3A and AARS), and antitumour ability in vitro and in vivo .Image 1