A high-throughput screen for the identification of compounds that inhibit nematode gene expression by targeting spliced leader trans-splicing.
Anthelminitics
Caenorhabditis elegans
HTS assay
SL1 trans-splicing
Sinefungin
Journal
International journal for parasitology. Drugs and drug resistance
ISSN: 2211-3207
Titre abrégé: Int J Parasitol Drugs Drug Resist
Pays: Netherlands
ID NLM: 101576715
Informations de publication
Date de publication:
08 2019
08 2019
Historique:
received:
12
01
2019
revised:
28
03
2019
accepted:
01
04
2019
pubmed:
25
4
2019
medline:
19
2
2020
entrez:
25
4
2019
Statut:
ppublish
Résumé
Infections with parasitic nematodes are among the most significant of the neglected tropical diseases affecting about a billion people living mainly in tropical regions with low economic activity. The most effective current strategy to control nematode infections involves large scale treatment programs with anthelmintic drugs. This strategy is at risk from the emergence of drug resistant parasites. Parasitic nematodes also affect livestock, which are treated with the same limited group of anthelmintic drugs. Livestock parasites resistant to single drugs, and even multi-drug resistant parasites, are appearing in many areas. There is therefore a pressing need for new anthelmintic drugs. Here we use the nematode Caenorhabditis elegans as a model for parasitic nematodes and demonstrate that sinefungin, a competitive inhibitor of methyltransferases, causes a delay in development and reduced fecundity, and inhibits spliced leader trans-splicing. Spliced leader trans-splicing is an essential step in gene expression that does not occur in the hosts of parasitic nematodes, and is therefore a potential target for new anthelmintic drugs. We have exploited the ability of sinefungin to inhibit spliced leader trans-splicing to adapt a green fluorescent protein based reporter gene assay that monitors spliced leader trans-splicing for high-throughput screening for new anthelmintic compounds. We have established a protocol for robust high-throughput screening, combining mechanical dispensing of living C. elegans into 384- or 1536- well plates with addition of compounds using an acoustic liquid dispenser, and the detection of the inhibition of SL trans-splicing using a microplate reader. We have tested this protocol in a first pilot screen and envisage that this assay will be a valuable tool in the search for new anthelmintic drugs.
Identifiants
pubmed: 31015150
pii: S2211-3207(18)30191-X
doi: 10.1016/j.ijpddr.2019.04.001
pmc: PMC6479105
pii:
doi:
Substances chimiques
Anthelmintics
0
RNA, Spliced Leader
0
Types de publication
Evaluation Study
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
28-37Subventions
Organisme : Medical Research Council
ID : MC_PC_14114
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/J007137/1
Pays : United Kingdom
Informations de copyright
Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.
Références
J Biomol Screen. 1999;4(2):67-73
pubmed: 10838414
RNA. 2007 Apr;13(4):511-20
pubmed: 17283210
Genetics. 2005 Nov;171(3):1033-45
pubmed: 16079230
WormBook. 2007 Mar 19;:1-15
pubmed: 18050497
Parasit Vectors. 2013 May 27;6:153
pubmed: 23711194
WormBook. 2006 Feb 11;:1-11
pubmed: 18050451
Int J Parasitol Drugs Drug Resist. 2017 Dec;7(3):262-271
pubmed: 28697451
Biol Psychiatry. 2013 Mar 1;73(5):464-71
pubmed: 23140663
Dev Biol. 2008 May 15;317(2):549-59
pubmed: 18395708
Wiley Interdiscip Rev RNA. 2011 May-Jun;2(3):417-34
pubmed: 21957027
PLoS One. 2013;8(2):e56877
pubmed: 23451103
Parasitology. 2000;121 Suppl:S39-50
pubmed: 11386690
Proc Natl Acad Sci U S A. 1990 Nov;87(22):8879-83
pubmed: 2247461
J Cell Biol. 2007 Aug 27;178(5):733-40
pubmed: 17709427
FEMS Microbiol Lett. 1990 Jun 1;57(3):239-43
pubmed: 2210336
J Biol Chem. 1978 Jun 25;253(12):4075-7
pubmed: 659406
Dev Biol. 1983 Mar;96(1):189-96
pubmed: 6825952
Am J Trop Med Hyg. 1983 Jan;32(1):31-3
pubmed: 6130709
FEBS Lett. 1980 Dec 1;121(2):287-91
pubmed: 6970144
Vet Parasitol. 2012 May 4;186(1-2):70-8
pubmed: 22154968
Lancet. 2006 May 6;367(9521):1521-32
pubmed: 16679166
J Biol Chem. 2008 Nov 28;283(48):33147-54
pubmed: 18826947
Cell. 1987 Jun 19;49(6):753-61
pubmed: 3581169
RNA. 1996 Aug;2(8):735-45
pubmed: 8752084
Genetics. 2014 Aug;197(4):1201-11
pubmed: 24931407
Nucleic Acids Res. 2016 Feb 29;44(4):1483-95
pubmed: 26773057
Cell. 1988 Aug 12;54(4):533-9
pubmed: 3401926
Biochem Biophys Res Commun. 1979 Aug 13;89(3):919-24
pubmed: 486211
Nat Protoc. 2008;3(6):1101-8
pubmed: 18546601
J Biol Chem. 2000 Dec 8;275(49):38311-8
pubmed: 10973970
RNA. 2011 Feb;17(2):327-37
pubmed: 21156961
Int J Parasitol Drugs Drug Resist. 2011 Oct 14;1(1):14-27
pubmed: 24533260
Vet Parasitol. 2011 Dec 15;182(2-4):264-8
pubmed: 21680095
Genes Dev. 1996 Jun 15;10(12):1543-56
pubmed: 8666237
Mol Cell Biol. 1992 Nov;12(11):4844-51
pubmed: 1406666
J Cell Biol. 1981 Jul;90(1):7-17
pubmed: 7251677
J Am Chem Soc. 2012 Oct 31;134(43):18004-14
pubmed: 23043551
Genetics. 1974 May;77(1):71-94
pubmed: 4366476
Curr Biol. 2001 Dec 11;11(24):1990-4
pubmed: 11747828
Biochem Biophys Res Commun. 1978 Nov 14;85(1):371-6
pubmed: 217377
ACS Med Chem Lett. 2014 Jan 31;5(4):293-7
pubmed: 24900829
Mol Cell Biol. 1992 Apr;12(4):1652-62
pubmed: 1549118
Nat Methods. 2012 Jul;9(7):671-5
pubmed: 22930834
Mol Biochem Parasitol. 2002 Jun;122(1):105-10
pubmed: 12076776
Trop Med Parasitol. 1986 Sep;37(3):255-7
pubmed: 2878488
Parasit Vectors. 2014 Dec 17;7:598
pubmed: 25515711
J Biol Chem. 1991 Dec 5;266(34):22792-5
pubmed: 1835972
J Vis Exp. 2011 May 19;(51):
pubmed: 21633332
Nucleic Acids Res. 2017 Aug 21;45(14):8474-8483
pubmed: 28582530