Application of RNAi to Genomic Drug Target Validation in Schistosomes.
Concerns over the possibility of resistance developing to praziquantel (PZQ), has stimulated efforts to develop new drugs for schistosomiasis. In addition to the development of improved whole organism screens, the success of RNA interference (RNAi) in schistosomes offers great promise for the identi...
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ftdoajarticles:oai:doaj.org/article:bfdc209395c44e1db0687cd2d05805dc 2023-05-15T15:18:32+02:00 Application of RNAi to Genomic Drug Target Validation in Schistosomes. Alessandra Guidi Nuha R Mansour Ross A Paveley Ian M Carruthers Jérémy Besnard Andrew L Hopkins Ian H Gilbert Quentin D Bickle 2015-05-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0003801 https://doaj.org/article/bfdc209395c44e1db0687cd2d05805dc EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC4438872?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0003801 https://doaj.org/article/bfdc209395c44e1db0687cd2d05805dc PLoS Neglected Tropical Diseases, Vol 9, Iss 5, p e0003801 (2015) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2015 ftdoajarticles https://doi.org/10.1371/journal.pntd.0003801 2022-12-31T04:48:53Z Concerns over the possibility of resistance developing to praziquantel (PZQ), has stimulated efforts to develop new drugs for schistosomiasis. In addition to the development of improved whole organism screens, the success of RNA interference (RNAi) in schistosomes offers great promise for the identification of potential drug targets to initiate drug discovery. In this study we set out to contribute to RNAi based validation of putative drug targets. Initially a list of 24 target candidates was compiled based on the identification of putative essential genes in schistosomes orthologous of C. elegans essential genes. Knockdown of Calmodulin (Smp_026560.2) (Sm-Calm), that topped this list, produced a phenotype characterised by waves of contraction in adult worms but no phenotype in schistosomula. Knockdown of the atypical Protein Kinase C (Smp_096310) (Sm-aPKC) resulted in loss of viability in both schistosomula and adults and led us to focus our attention on other kinase genes that were identified in the above list and through whole organism screening of known kinase inhibitor sets followed by chemogenomic evaluation. RNAi knockdown of these kinase genes failed to affect adult worm viability but, like Sm-aPKC, knockdown of Polo-like kinase 1, Sm-PLK1 (Smp_009600) and p38-MAPK, Sm-MAPK p38 (Smp_133020) resulted in an increased mortality of schistosomula after 2-3 weeks, an effect more marked in the presence of human red blood cells (hRBC). For Sm-PLK-1 the same effects were seen with the specific inhibitor, BI2536, which also affected viable egg production in adult worms. For Sm-PLK-1 and Sm-aPKC the in vitro effects were reflected in lower recoveries in vivo. We conclude that the use of RNAi combined with culture with hRBC is a reliable method for evaluating genes important for larval development. However, in view of the slow manifestation of the effects of Sm-aPKC knockdown in adults and the lack of effects of Sm-PLK-1 and Sm-MAPK p38 on adult viability, these kinases may not represent suitable drug targets. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Polo ENVELOPE(28.967,28.967,65.600,65.600) PLOS Neglected Tropical Diseases 9 5 e0003801 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Alessandra Guidi Nuha R Mansour Ross A Paveley Ian M Carruthers Jérémy Besnard Andrew L Hopkins Ian H Gilbert Quentin D Bickle Application of RNAi to Genomic Drug Target Validation in Schistosomes. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Concerns over the possibility of resistance developing to praziquantel (PZQ), has stimulated efforts to develop new drugs for schistosomiasis. In addition to the development of improved whole organism screens, the success of RNA interference (RNAi) in schistosomes offers great promise for the identification of potential drug targets to initiate drug discovery. In this study we set out to contribute to RNAi based validation of putative drug targets. Initially a list of 24 target candidates was compiled based on the identification of putative essential genes in schistosomes orthologous of C. elegans essential genes. Knockdown of Calmodulin (Smp_026560.2) (Sm-Calm), that topped this list, produced a phenotype characterised by waves of contraction in adult worms but no phenotype in schistosomula. Knockdown of the atypical Protein Kinase C (Smp_096310) (Sm-aPKC) resulted in loss of viability in both schistosomula and adults and led us to focus our attention on other kinase genes that were identified in the above list and through whole organism screening of known kinase inhibitor sets followed by chemogenomic evaluation. RNAi knockdown of these kinase genes failed to affect adult worm viability but, like Sm-aPKC, knockdown of Polo-like kinase 1, Sm-PLK1 (Smp_009600) and p38-MAPK, Sm-MAPK p38 (Smp_133020) resulted in an increased mortality of schistosomula after 2-3 weeks, an effect more marked in the presence of human red blood cells (hRBC). For Sm-PLK-1 the same effects were seen with the specific inhibitor, BI2536, which also affected viable egg production in adult worms. For Sm-PLK-1 and Sm-aPKC the in vitro effects were reflected in lower recoveries in vivo. We conclude that the use of RNAi combined with culture with hRBC is a reliable method for evaluating genes important for larval development. However, in view of the slow manifestation of the effects of Sm-aPKC knockdown in adults and the lack of effects of Sm-PLK-1 and Sm-MAPK p38 on adult viability, these kinases may not represent suitable drug targets. |
format |
Article in Journal/Newspaper |
author |
Alessandra Guidi Nuha R Mansour Ross A Paveley Ian M Carruthers Jérémy Besnard Andrew L Hopkins Ian H Gilbert Quentin D Bickle |
author_facet |
Alessandra Guidi Nuha R Mansour Ross A Paveley Ian M Carruthers Jérémy Besnard Andrew L Hopkins Ian H Gilbert Quentin D Bickle |
author_sort |
Alessandra Guidi |
title |
Application of RNAi to Genomic Drug Target Validation in Schistosomes. |
title_short |
Application of RNAi to Genomic Drug Target Validation in Schistosomes. |
title_full |
Application of RNAi to Genomic Drug Target Validation in Schistosomes. |
title_fullStr |
Application of RNAi to Genomic Drug Target Validation in Schistosomes. |
title_full_unstemmed |
Application of RNAi to Genomic Drug Target Validation in Schistosomes. |
title_sort |
application of rnai to genomic drug target validation in schistosomes. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2015 |
url |
https://doi.org/10.1371/journal.pntd.0003801 https://doaj.org/article/bfdc209395c44e1db0687cd2d05805dc |
long_lat |
ENVELOPE(28.967,28.967,65.600,65.600) |
geographic |
Arctic Polo |
geographic_facet |
Arctic Polo |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 9, Iss 5, p e0003801 (2015) |
op_relation |
http://europepmc.org/articles/PMC4438872?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0003801 https://doaj.org/article/bfdc209395c44e1db0687cd2d05805dc |
op_doi |
https://doi.org/10.1371/journal.pntd.0003801 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
9 |
container_issue |
5 |
container_start_page |
e0003801 |
_version_ |
1766348733592633344 |