Strategies for In Vivo Screening and Mitigation of Hepatotoxicity Associated with Antisense Drugs

Piotr J. Kamola*, Klio Maratou, Paul A. Wilson, Kay Rush, Tanya Mullaney, Tom McKevitt, Paula Evans, Jim Ridings, Probash Chowdhury, Aude Roulois, Ann Fairchild, Sean McCawley, Karen Cartwright, Nigel J. Gooderham, Timothy Gant, Kitty Moores, Stephen A. Hughes, Mark R. Edbrooke, Kenneth Clark, Joel D. Parry

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

34 Citations (Scopus)


Antisense oligonucleotide (ASO) gapmers downregulate gene expression by inducing enzyme-dependent degradation of targeted RNA and represent a promising therapeutic platform for addressing previously undruggable genes. Unfortunately, their therapeutic application, particularly that of the more potent chemistries (e.g., locked-nucleic-acid-containing gapmers), has been hampered by their frequent hepatoxicity, which could be driven by hybridization-mediated interactions. An early de-risking of this liability is a crucial component of developing safe, ASO-based drugs. To rank ASOs based on their effect on the liver, we have developed an acute screen in the mouse that can be applied early in the drug development cycle. A single-dose (3-day) screen with streamlined endpoints (i.e., plasma transaminase levels and liver weights) was observed to be predictive of ASO hepatotoxicity ranking established based on a repeat-dose (15 day) study. Furthermore, to study the underlying mechanisms of liver toxicity, we applied transcriptome profiling and pathway analyses and show that adverse in vivo liver phenotypes correlate with the number of potent, hybridization-mediated off-target effects (OTEs). We propose that a combination of in silico OTE predictions, streamlined in vivo hepatotoxicity screening, and a transcriptome-wide selectivity screen is a valid approach to identifying and progressing safer compounds.

Original languageEnglish
Pages (from-to)383-394
Number of pages12
JournalMolecular Therapy - Nucleic Acids
Publication statusPublished - 15 Sept 2017

Bibliographical note

Funding Information:
The project was supported by a EPSRC Industrial CASE Award ( Imperial College London, GlaxoSmithKline and Public Health England ; EP/J502017/1 ) to P.J.K. Analysis and manuscript preparation were finalized at the Laboratory for Medical Science Mathematics, RIKEN Center for Integrative Medical Sciences in Yokohama, Japan, under T. Tsunoda and supported by a JSPS postdoctoral fellowship ( 15F15776 ) and a JST CREST grant ( JPMJCR1412 ). Funding for the open access charge was provided by GlaxoSmithKline . The authors would like to thank the anonymous reviewers who provided very helpful suggestions and comments.

Publisher Copyright:
© 2017 GSK R&D


  • ASOs
  • LNA
  • OTEs
  • RNArcher
  • antisense oligonucleotides
  • gene silencing
  • hepatotoxicity
  • locked nucleic acids
  • off-target effects
  • selectivity


Dive into the research topics of 'Strategies for In Vivo Screening and Mitigation of Hepatotoxicity Associated with Antisense Drugs'. Together they form a unique fingerprint.

Cite this