Systemic delivery of messenger RNA for the treatment of pancreatic cancer using polyplex nanomicelles with a cholesterol moiety

Satoshi Uchida; Hiroaki Kinoh; Takehiko Ishii; Akitsugu Matsui; Theofilus Agrios Tockary; Kaori Machitani Takeda; Hirokuni Uchida; Kensuke Osada; Keiji Itaka; Kazunori Kataoka

doi:10.1016/j.biomaterials.2015.12.031

Systemic delivery of messenger RNA for the treatment of pancreatic cancer using polyplex nanomicelles with a cholesterol moiety

Biomaterials. 2016 Mar:82:221-8. doi: 10.1016/j.biomaterials.2015.12.031. Epub 2015 Dec 31.

Affiliations

¹ Laboratory of Clinical Biotechnology, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
² Department of Bioengineering, Graduate School of Engineering, The University of Tokyo, Tokyo, Japan.
³ Department of Materials Engineering, Graduate School of Engineering, The University of Tokyo, Tokyo, Japan.
⁴ Laboratory of Clinical Biotechnology, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan. Electronic address: itaka-ort@umin.net.
⁵ Laboratory of Clinical Biotechnology, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan; Department of Bioengineering, Graduate School of Engineering, The University of Tokyo, Tokyo, Japan; Department of Materials Engineering, Graduate School of Engineering, The University of Tokyo, Tokyo, Japan. Electronic address: kataoka@bmw.t.u-tokyo.ac.jp.

PMID: 26763736
DOI: 10.1016/j.biomaterials.2015.12.031

Abstract

Systemic delivery of messenger RNA (mRNA) is technically challenging because mRNA is highly susceptible to enzymatic degradation in the blood circulation. In this study, we used a nanomicelle-based platform, prepared from mRNA and poly(ethylene glycol) (PEG)-polycation block copolymers. A cholesterol (Chol) moiety was attached to the ω-terminus of the block copolymer to increase the stability of the nanomicelle by hydrophobic interaction. After in vitro screening, polyaspartamide with four aminoethylene repeats in its side chain (PAsp(TEP)) was selected as the cationic segment of the block copolymer, because it contributes to enhance nuclease resistance and high protein expression from the mRNA. After intravenous injection, PEG-PAsp(TEP)-Chol nanomicelles showed significantly enhanced blood retention of mRNA in comparison to nanomicelles without Chol. We used the nanomicelles for treating intractable pancreatic cancer in a subcutaneous inoculation mouse model through the delivery of mRNA encoding an anti-angiogenic protein (sFlt-1). PEG-PAsp(TEP)-Chol nanomicelles generated efficient protein expression from the delivered mRNA in tumor tissue, resulting in remarkable inhibition of the tumor growth, whereas nanomicelles without Chol failed to show a detectable therapeutic effect. In conclusion, the stabilized nanomicelle system led to the successful systemic delivery of mRNA in therapeutic application, holding great promise for the treatment of various diseases.

Keywords: Anti-angiogenic therapy; Cholesterol; Enzymatic degradation; Pancreatic cancer; Polyplex nanomicelle; mRNA delivery.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis / drug effects
Apoptosis / genetics
Cell Line, Tumor
Cholesterol / chemistry*
Female
Genetic Therapy / methods*
Humans
Hydrophobic and Hydrophilic Interactions
Mice
Mice, Inbred BALB C
Micelles
Nanocapsules / administration & dosage
Nanocapsules / chemistry*
Nanocapsules / ultrastructure
Pancreatic Neoplasms / genetics
Pancreatic Neoplasms / pathology*
Pancreatic Neoplasms / therapy*
Particle Size
RNA, Messenger / administration & dosage*
Treatment Outcome

Substances

Micelles
Nanocapsules
RNA, Messenger
Cholesterol