Advertisement

In Vivo Administration of Therapeutic Antisense Oligonucleotides

  • Luisa Statello
  • Mohamad Moustafa Ali
  • Chandrasekhar KanduriEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2254)

Abstract

With the rapid revolution in RNA/DNA sequencing technologies, it is evident that mammalian genomes express tens of thousands of long noncoding RNAs (lncRNAs). Since a large majority of lncRNAs have been functionally implicated in cancer development and progression, there is an increasing appreciation for the use of antisense oligonucleotide (ASO)-based therapies targeting lncRNAs in several cancers. Despite their great potential in therapeutic applications, their use is still limited due to cellular toxicity and shortcomings in achieving required stability in biological fluids and tissue uptake. To overcome these limitations, major changes in ASO chemistry have been introduced to generate second and third generation ASOs, including locked nucleic acids (LNA) technology. Here we describe two different LNA-ASO delivery approaches, a peritumoral administration and a systemic delivery in xenograft models of lung adenocarcinoma, that significantly reduced tumor growth without inducing toxicity.

Key words

Antisense oligonucleotides LNA-ASOs Xenografts Lung cancer lncRNAs Long noncoding RNAs 

Notes

Acknowledgments

This work was supported by the grants from Knut and Alice Wallenberg Foundation [KAW2014.0057], Swedish Foundation for Strategic Research [RB13-0204], Swedish Cancer Research foundation [Cancerfonden: Kontrakt no. CAN2018/591], Swedish Research Council [2017-02834], Barncancerfonden [PR2018-0090], Ingabritt Och Arne Lundbergs forskningsstiftelse, and LUA/ALF (to C.K.).

References

  1. 1.
    Dias N, Stein CA (2002) Antisense oligonucleotides: basic concepts and mechanisms. Mol Cancer Ther 1:347–355CrossRefGoogle Scholar
  2. 2.
    Schoch KM, Miller TM (2017) Antisense oligonucleotides: translation from mouse models to human neurodegenerative diseases. Neuron 94:1056–1070CrossRefGoogle Scholar
  3. 3.
    Crooke ST (2017) Molecular mechanisms of antisense oligonucleotides. Nucleic Acid Ther 27:70–77CrossRefGoogle Scholar
  4. 4.
    Havens MA, Hastings ML (2016) Splice-switching antisense oligonucleotides as therapeutic drugs. Nucleic Acids Res 44:6549–6563CrossRefGoogle Scholar
  5. 5.
    Ling H, Fabbri M, Calin GA (2013) MicroRNAs and other non-coding RNAs as targets for anticancer drug development. Nat Rev Drug Discov 12:847–865CrossRefGoogle Scholar
  6. 6.
    Liang XH, Sun H, Nichols JG et al (2017) RNase H1-dependent antisense oligonucleotides are robustly active in directing RNA cleavage in both the cytoplasm and the nucleus. Mol Ther 25:2075–2092CrossRefGoogle Scholar
  7. 7.
    Erickson MA, Niehoff ML, Farr SA et al (2012) Peripheral administration of antisense oligonucleotides targeting the amyloid-beta protein precursor reverses AbetaPP and LRP-1 overexpression in the aged SAMP8 mouse brain. J Alzheimers Dis 28:951–960CrossRefGoogle Scholar
  8. 8.
    Viereck J, Kumarswamy R, Foinquinos A et al (2016) Long noncoding RNA Chast promotes cardiac remodeling. Sci Transl Med 8:326ra322CrossRefGoogle Scholar
  9. 9.
    Bremmer-Bout M, Aartsma-Rus A, De Meijer EJ et al (2004) Targeted exon skipping in transgenic hDMD mice: a model for direct preclinical screening of human-specific antisense oligonucleotides. Mol Ther 10:232–240CrossRefGoogle Scholar
  10. 10.
    Cao C, Mu Y, Hallahan DE et al (2004) XIAP and survivin as therapeutic targets for radiation sensitization in preclinical models of lung cancer. Oncogene 23:7047–7052CrossRefGoogle Scholar
  11. 11.
    Stein CA, Castanotto D (2017) FDA-approved oligonucleotide therapies in 2017. Mol Ther 25:1069–1075CrossRefGoogle Scholar
  12. 12.
    Rinaldi C, Wood MJA (2018) Antisense oligonucleotides: the next frontier for treatment of neurological disorders. Nat Rev Neurol 14:9–21CrossRefGoogle Scholar
  13. 13.
    Reilley MJ, Mccoon P, Cook C et al (2018) STAT3 antisense oligonucleotide AZD9150 in a subset of patients with heavily pretreated lymphoma: results of a phase 1b trial. J Immunother Cancer 6:119CrossRefGoogle Scholar
  14. 14.
    Kamola PJ, Kitson JD, Turner G et al (2015) In silico and in vitro evaluation of exonic and intronic off-target effects form a critical element of therapeutic ASO gapmer optimization. Nucleic Acids Res 43:8638–8650CrossRefGoogle Scholar
  15. 15.
    Agrawal S, Kandimalla ER (2004) Role of toll-like receptors in antisense and siRNA [corrected]. Nat Biotechnol 22:1533–1537CrossRefGoogle Scholar
  16. 16.
    Yoshida T, Naito Y, Sasaki K et al (2018) Estimated number of off-target candidate sites for antisense oligonucleotides in human mRNA sequences. Genes Cells 23:448–455CrossRefGoogle Scholar
  17. 17.
    Ali MM, Akhade VS, Kosalai ST et al (2018) PAN-cancer analysis of S-phase enriched lncRNAs identifies oncogenic drivers and biomarkers. Nat Commun 9:883CrossRefGoogle Scholar
  18. 18.
    Leucci E, Vendramin R, Spinazzi M et al (2016) Melanoma addiction to the long non-coding RNA SAMMSON. Nature 531:518–522CrossRefGoogle Scholar
  19. 19.
    Michalik KM, You X, Manavski Y et al (2014) Long noncoding RNA MALAT1 regulates endothelial cell function and vessel growth. Circ Res 114:1389–1397CrossRefGoogle Scholar
  20. 20.
    Straarup EM, Fisker N, Hedtjarn M et al (2010) Short locked nucleic acid antisense oligonucleotides potently reduce apolipoprotein B mRNA and serum cholesterol in mice and non-human primates. Nucleic Acids Res 38:7100–7111CrossRefGoogle Scholar
  21. 21.
    Starckx S, Batheja A, Verheyen GR et al (2013) Evaluation of miR-122 and other biomarkers in distinct acute liver injury in rats. Toxicol Pathol 41:795–804CrossRefGoogle Scholar
  22. 22.
    Sharapova T, Devanarayan V, Leroy B et al (2016) Evaluation of miR-122 as a serum biomarker for hepatotoxicity in investigative rat toxicology studies. Vet Pathol 53:211–221CrossRefGoogle Scholar
  23. 23.
    Burel SA, Han SR, Lee HS et al (2013) Preclinical evaluation of the toxicological effects of a novel constrained ethyl modified antisense compound targeting signal transducer and activator of transcription 3 in mice and cynomolgus monkeys. Nucleic Acid Ther 23:213–227CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  • Luisa Statello
    • 1
  • Mohamad Moustafa Ali
    • 1
  • Chandrasekhar Kanduri
    • 1
    Email author
  1. 1.Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska AcademyUniversity of GothenburgGothenburgSweden

Personalised recommendations