論文

国際誌
2017年9月20日

Systemic Delivery of Folate-PEG siRNA Lipopolyplexes with Enhanced Intracellular Stability for In Vivo Gene Silencing in Leukemia.

Bioconjugate chemistry
  • Dian-Jang Lee
  • Eva Kessel
  • Taavi Lehto
  • Xueying Liu
  • Naoto Yoshinaga
  • Kärt Padari
  • Ying-Chen Chen
  • Susanne Kempter
  • Satoshi Uchida
  • Joachim O Rädler
  • Margus Pooga
  • Ming-Thau Sheu
  • Kazunori Kataoka
  • Ernst Wagner
  • 全て表示

28
9
開始ページ
2393
終了ページ
2409
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1021/acs.bioconjchem.7b00383

Protection of small interfering RNA (siRNA) against degradation and targeted delivery across the plasma and endosomal membranes to the final site of RNA interference (RNAi) are major aims for the development of siRNA therapeutics. Targeting for folate receptor (FR)-expressing tumors, we optimized siRNA polyplexes by coformulating a folate-PEG-oligoaminoamide (for surface shielding and targeting) with one of three lipo-oligoaminoamides (optionally tyrosine-modified, for optimizing stability and size) to generate ∼100 nm targeted lipopolyplexes (TLPs), which self-stabilize by cysteine disulfide cross-links. To better understand parameters for improved tumor-directed gene silencing, we analyzed intracellular distribution and siRNA release kinetics. FR-mediated endocytosis and endosomal escape of TLPs was confirmed by immuno-TEM. We monitored colocalization of TLPs with endosomes and lysosomes, and onset of siRNA release by time-lapse confocal microscopy; analyzed intracellular stability by FRET using double-labeled siRNA; and correlated results with knockdown of eGFPLuc protein and EG5 mRNA expression. The most potent formulation, TLP1, containing lipopolyplex-stabilizing tyrosine trimers, was found to unpack siRNA in sustained manner with up to 5-fold higher intracellular siRNA stability after 4 h compared to other TLPs. Unexpectedly, data indicated that intracellular siRNA stability instead of an early endosomal exit dominate as a deciding factor for silencing efficiency of TLPs. After i.v. administration in a subcutaneous leukemia mouse model, TLP1 exhibited ligand-dependent tumoral siRNA retention, resulting in 65% EG5 gene silencing at mRNA level without detectable adverse effects. In sum, tyrosine-modified TLP1 conveys superior protection of siRNA for an effective tumor-targeted delivery and RNAi in vivo.

リンク情報
DOI
https://doi.org/10.1021/acs.bioconjchem.7b00383
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/28772071
ID情報
  • DOI : 10.1021/acs.bioconjchem.7b00383
  • PubMed ID : 28772071

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