TY - JOUR
T1 - 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
AU - Ma, Yuan
AU - Zhao, Wenting
AU - Li, Yiding
AU - Pan, Yufei
AU - Wang, Shuhe
AU - Zhu, Yuejie
AU - Kong, Lingxuan
AU - Guan, Zhu
AU - Wang, Jiancheng
AU - Zhang, Lihe
AU - Yang, Zhenjun
N1 - Funding information:
We thank Cnkingbio Company Ltd. (Beijing, China) for transcriptome and proteomic microarray support. This work was supported by the Ministry of Science and Technology of China (Grant No. 2017ZX09303013) and the National Natural Science Foundation of China (Grant No. 21778006, 21332010).
Publisher copyright:
© 2019 Elsevier Ltd.
PY - 2019/3
Y1 - 2019/3
N2 - 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.
AB - 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.
KW - Gene delivery
KW - G3139
KW - DNCA
KW - PS modification
KW - LNA-gapmer
UR - https://www.scopus.com/inward/record.uri?eid=2-s2.0-85060925581&doi=10.1016%2fj.biomaterials.2018.12.033&partnerID=40&md5=16e1c068e6a0a4fa10c32108309ea56f
U2 - 10.1016/j.biomaterials.2018.12.033
DO - 10.1016/j.biomaterials.2018.12.033
M3 - Journal article
SN - 0142-9612
VL - 197
SP - 182
EP - 193
JO - Biomaterials
JF - Biomaterials
ER -