Sage Journals: Discover world-class research

Abstract

Antisense oligonucleotides (ASOs) are classified into gapmer and non-gapmer types according to their chemical modification pattern and mechanism of action. Although gapmer ASOs effectively reduce target RNA expression through intracellular RNase H1, high-affinity gapmer ASOs also have hepatotoxic potential. Non-gapmer ASOs, which are mainly used for pre-mRNA splicing regulation or functional inhibition of microRNA through their steric effects, are also able to inhibit target RNA expression using nonsense-mediated decay. However, it was unknown if they induce high knockdown activity without showing hepatotoxicity. In this study, we investigated the modification pattern of non-gapmer ASOs and show that they have comparable knockdown potential if they have an appropriate melting temperature (T_m) range. We also demonstrated that non-gapmer ASOs show high knockdown effects without inducing hepatotoxicity in the mouse liver. These results indicated that non-gapmer ASOs have the potential to become an alternative inhibitor of target expression with a lower risk of hepatotoxicity.

Get full access to this article

View all access options for this article.

References

Crooke

. (2017). Molecular mechanisms of antisense oligonucleotides. Nucleic Acid Ther, 27:70–77.

Khvorova

and Watts

. (2017). The chemical evolution of oligonucleotide therapies of clinical utility. Nat Biotechnol, 35:238–248.

Yamamoto

, Nakatani

, Narukawa

and Obika

. (2011). Antisense drug discovery and development. Future Med Chem, 3:339–365.

Crooke

, Wang

, Vickers

, Shen

and Liang

. (2017). Cellular uptake and trafficking of antisense oligonucleotides. Nat Biotechnol, 35:230–237.

Bennett

and Swayze

. (2010). RNA targeting therapeutics: molecular mechanisms of antisense oligonucleotides as a therapeutic platform. Annu Rev Pharmacol Toxicol, 50:259–293.

FDA. (2013). FDA Approves New Orphan Drug Kynamro to Treat Inherited Cholesterol Disorder. . FDA Press Announc.

Wong

and Goldberg

. (2014). Mipomersen (kynamro): a novel antisense oligonucleotide inhibitor for the management of homozygous familial hypercholesterolemia. P T, 39:119–122.

Singh

, Koshkin

, Wengel

and Nielsen

. (1998). LNA (locked nucleic acids): synthesis and high-affinity nucleic acid recognition. Chem Commun. 455–456.

Obika

, Nanbu

, Hari

, Morio

, In

, Ishida

and Imanishi

. (1997). Synthesis of 2′-O,4′-C-methyleneuridine and -cytidine. Novel bicyclic nucleosides having a fixed C3-endo sugar puckering. Tetrahedron Lett, 38:8735–8738.

10.

Obika

, Nanbu

, Hari

, Andoh

, Morio

, Doi

and Imanishi

. (1998). Stability and structural features of the duplexes containing nucleoside analogues with a fixed N-type conformation, 2′-O,4′-C-methyleneribonucleosides. Tetrahedron Lett, 39:5401–5404.

11.

Swayze

, Siwkowski

, Wancewicz

, Migawa

, Wyrzykiewicz

, Hung

, Monia

and Bennett

. (2007). Antisense oligonucleotides containing locked nucleic acid improve potency but cause significant hepatotoxicity in animals. Nucleic Acids Res, 35:687–700.

12.

Kasuya

, Hori

, Watanabe

, Nakajima

, Gahara

, Rokushima

, Yanagimoto

and Kugimiya

. (2016). Ribonuclease H1-dependent hepatotoxicity caused by locked nucleic acid-modified gapmer antisense oligonucleotides. Sci Rep, 6:30377.

13.

Burel

, Hart

, Cauntay

, Hsiao

, Machemer

, Katz

, Watt

, Bui

, Younis

, et al. (2016). Hepatotoxicity of high affinity gapmer antisense oligonucleotides is mediated by RNase H1 dependent promiscuous reduction of very long pre-mRNA transcripts. Nucleic Acids Res, 44:2093–2109.

14.

Stein

and Castanotto

. (2017). FDA-approved oligonucleotide therapies in 2017. Mol Ther, 25:1069–1075.

15.

Ward

, Norrbom

, Chun

, Bennett

and Rigo

. (2014). Nonsense-mediated decay as a terminating mechanism for antisense oligonucleotides. Nucleic Acids Res, 42:5871–5879.

16.

Yahara

, Shrestha

, Yamamoto

, Hari

, Osawa

, Yamaguchi

, Nishida

, Kodama

and Obika

. (2012). Amido-bridged nucleic acids (AmNAs): synthesis, duplex stability, nuclease resistance, and in vitro antisense potency. Chembiochem, 13:2513–2516.

17.

Miyashita

, Rahman

SMA

, Seki

, Obika

and Imanishi

. (2007). N-methyl substituted 2′,4′-BNANC: a highly nuclease-resistant nucleic acid analogue with high-affinity RNA selective hybridization. Chem Commun (Camb) 3765–3767.

18.

Mitsuoka

, Kodama

, Ohnishi

, Hari

, Imanishi

and Obika

. (2009). A bridged nucleic acid, 2′, 4′-BNACOC: synthesis of fully modified oligonucleotides bearing thymine, 5-methylcytosine, adenine and guanine 2′, 4′-BNACOCmonomers and RNA-selective nucleic-acid recognition. Nucleic Acids Res, 37:1225–1238.

19.

Hari

, Obika

, Ohnishi

, Eguchi

, Osaki

, Ohishi

and Imanishi

. (2006). Synthesis and properties of 2′-O,4′-C-methyleneoxymethylene bridged nucleic acid. Bioorg Med Chem, 14:1029–1038.

20.

Yamamoto

, Yahara

, Waki

, Yasuhara

, Wada

, Harada-Shiba

and Obika

. (2015). Amido-bridged nucleic acids with small hydrophobic residues enhance hepatic tropism of antisense oligonucleotides in vivo. Org Biomol Chem, 13:3757–3765.

21.

Shimo

, Tachibana

, Saito

, Yoshida

, Tomita

, Waki

, Yamamoto

, Doi

, Inoue

, Kawakami

and Obika

. (2014). Design and evaluation of locked nucleic acid-based splice-switching oligonucleotides in vitro. Nucleic Acids Res, 42:8174–8187.

22.

Zammarchi

, de Stanchina

, Bournazou

, Supakorndej

, Martires

, Riedel

, Corben

, Bromberg

and Cartegni

. (2011). Antitumorigenic potential of STAT3 alternative splicing modulation. Proc Natl Acad Sci U S A, 108:17779–17784.

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

3.89 MB

0.04 MB

0.03 MB

0.07 MB

0.11 MB

0.15 MB

RNA Reduction and Hepatotoxic Potential Caused by Non-Gapmer Antisense Oligonucleotides

Abstract

Get full access to this article

References

Supplementary Material