Imperial College London
Alternate

Emeritus ProfessorNigelGooderham

Faculty of Medicine, Department of Metabolism, Digestion and Reproduction

Emeritus Professor of Molecular Toxicology
 
 
 
//

Contact

 

n.gooderham Website

 
 
//

Location

 

Burlington DanesHammersmith Campus

//

Summary

 

Publications

Citation

BibTex format

@article{Kamola:2015:nar/gkv857,
author = {Kamola, PJ and Kitson, JD and Turner, G and Maratou, K and Eriksson, S and Panjwani, A and Warnock, LC and Douillard, Guilloux GA and Moores, K and Koppe, EL and Wixted, WE and Wilson, PA and Gooderham, NJ and Gant, TW and Clark, KL and Hughes, SA and Edbrooke, MR and Parry, JD},
doi = {nar/gkv857},
journal = {Nucleic Acids Research},
pages = {8638--8650},
title = {In silico and in vitro evaluation of exonic and intronic off-target effects form a critical element of therapeutic ASO gapmer optimization.},
url = {http://dx.doi.org/10.1093/nar/gkv857},
volume = {43},
year = {2015}
}
Download

RIS format (EndNote, RefMan)

TY  - JOUR
AB  - With many safety and technical limitations partly mitigated through chemical modifications, antisense oligonucleotides (ASOs) are gaining recognition as therapeutic entities. The increase in potency realized by 'third generation chemistries' may, however, simultaneously increase affinity to unintended targets with partial sequence complementarity. However, putative hybridization-dependent off-target effects (OTEs), a risk historically regarded as low, are not being adequately investigated. Here we show an unexpectedly high OTEs confirmation rate during screening of fully phosphorothioated (PS)-LNA gapmer ASOs designed against the BACH1 transcript. We demonstrate in vitro mRNA and protein knockdown of off-targets with a wide range of mismatch (MM) and gap patterns. Furthermore, with RNase H1 activity residing within the nucleus, hybridization predicted against intronic regions of pre-mRNAs was tested and confirmed. This dramatically increased ASO-binding landscape together with relatively high potency of such interactions translates into a considerable safety concern. We show here that with base pairing-driven target recognition it is possible to predict the putative off-targets and address the liability during lead design and optimization phases. Moreover, in silico analysis performed against both primary as well as spliced transcripts will be invaluable in elucidating the mechanism behind the hepatoxicity observed with some LNA-modified gapmers.
AU  - Kamola,PJ
AU  - Kitson,JD
AU  - Turner,G
AU  - Maratou,K
AU  - Eriksson,S
AU  - Panjwani,A
AU  - Warnock,LC
AU  - Douillard,Guilloux GA
AU  - Moores,K
AU  - Koppe,EL
AU  - Wixted,WE
AU  - Wilson,PA
AU  - Gooderham,NJ
AU  - Gant,TW
AU  - Clark,KL
AU  - Hughes,SA
AU  - Edbrooke,MR
AU  - Parry,JD
DO  - nar/gkv857
EP  - 8650
PY  - 2015///
SN  - 1362-4962
SP  - 8638
TI  - In silico and in vitro evaluation of exonic and intronic off-target effects form a critical element of therapeutic ASO gapmer optimization.
T2  - Nucleic Acids Research
UR  - http://dx.doi.org/10.1093/nar/gkv857
UR  - http://hdl.handle.net/10044/1/26355
VL  - 43
ER  -
Download