Cloning and functional complementation of ten Schistosoma mansoni phosphodiesterases expressed in the mammalian host stages.
Only a single drug against schistosomiasis is currently available and new drug development is urgently required but very few drug targets have been validated and characterised. However, regulatory systems including cyclic nucleotide metabolism are emerging as primary candidates for drug discovery. H...
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ftdoajarticles:oai:doaj.org/article:dec44c4ba15a44afbe2fcc56f7a87eca 2023-05-15T15:10:58+02:00 Cloning and functional complementation of ten Schistosoma mansoni phosphodiesterases expressed in the mammalian host stages. Jane C Munday Stefan Kunz Titilola D Kalejaiye Marco Siderius Susanne Schroeder Daniel Paape Ali H Alghamdi Zainab Abbasi Sheng Xiang Huang Anne-Marie Donachie Samia William Abdel Nasser Sabra Geert Jan Sterk Sanaa S Botros David G Brown Charles S Hoffman Rob Leurs Harry P de Koning 2020-07-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0008447 https://doaj.org/article/dec44c4ba15a44afbe2fcc56f7a87eca EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0008447 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0008447 https://doaj.org/article/dec44c4ba15a44afbe2fcc56f7a87eca PLoS Neglected Tropical Diseases, Vol 14, Iss 7, p e0008447 (2020) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2020 ftdoajarticles https://doi.org/10.1371/journal.pntd.0008447 2022-12-31T07:46:37Z Only a single drug against schistosomiasis is currently available and new drug development is urgently required but very few drug targets have been validated and characterised. However, regulatory systems including cyclic nucleotide metabolism are emerging as primary candidates for drug discovery. Here, we report the cloning of ten cyclic nucleotide phosphodiesterase (PDE) genes of S. mansoni, out of a total of 11 identified in its genome. We classify these PDEs by homology to human PDEs. Male worms displayed higher expression levels for all PDEs, in mature and juvenile worms, and schistosomula. Several functional complementation approaches were used to characterise these genes. We constructed a Trypanosoma brucei cell line in which expression of a cAMP-degrading PDE complements the deletion of TbrPDEB1/B2. Inhibitor screens of these cells expressing only either SmPDE4A, TbrPDEB1 or TbrPDEB2, identified highly potent inhibitors of the S. mansoni enzyme that elevated the cellular cAMP concentration. We further expressed most of the cloned SmPDEs in two pde1Δ/pde2Δ strains of Saccharomyces cerevisiae and some also in a specialised strain of Schizosacharomyces pombe. Five PDEs, SmPDE1, SmPDE4A, SmPDE8, SmPDE9A and SmPDE11 successfully complemented the S. cerevisiae strains, and SmPDE7var also complemented to a lesser degree, in liquid culture. SmPDE4A, SmPDE8 and SmPDE11 were further assessed in S. pombe for hydrolysis of cAMP and cGMP; SmPDE11 displayed considerable preferrence for cGMP over cAMP. These results and tools enable the pursuit of a rigorous drug discovery program based on inhibitors of S. mansoni PDEs. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 14 7 e0008447 |
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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 Jane C Munday Stefan Kunz Titilola D Kalejaiye Marco Siderius Susanne Schroeder Daniel Paape Ali H Alghamdi Zainab Abbasi Sheng Xiang Huang Anne-Marie Donachie Samia William Abdel Nasser Sabra Geert Jan Sterk Sanaa S Botros David G Brown Charles S Hoffman Rob Leurs Harry P de Koning Cloning and functional complementation of ten Schistosoma mansoni phosphodiesterases expressed in the mammalian host stages. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Only a single drug against schistosomiasis is currently available and new drug development is urgently required but very few drug targets have been validated and characterised. However, regulatory systems including cyclic nucleotide metabolism are emerging as primary candidates for drug discovery. Here, we report the cloning of ten cyclic nucleotide phosphodiesterase (PDE) genes of S. mansoni, out of a total of 11 identified in its genome. We classify these PDEs by homology to human PDEs. Male worms displayed higher expression levels for all PDEs, in mature and juvenile worms, and schistosomula. Several functional complementation approaches were used to characterise these genes. We constructed a Trypanosoma brucei cell line in which expression of a cAMP-degrading PDE complements the deletion of TbrPDEB1/B2. Inhibitor screens of these cells expressing only either SmPDE4A, TbrPDEB1 or TbrPDEB2, identified highly potent inhibitors of the S. mansoni enzyme that elevated the cellular cAMP concentration. We further expressed most of the cloned SmPDEs in two pde1Δ/pde2Δ strains of Saccharomyces cerevisiae and some also in a specialised strain of Schizosacharomyces pombe. Five PDEs, SmPDE1, SmPDE4A, SmPDE8, SmPDE9A and SmPDE11 successfully complemented the S. cerevisiae strains, and SmPDE7var also complemented to a lesser degree, in liquid culture. SmPDE4A, SmPDE8 and SmPDE11 were further assessed in S. pombe for hydrolysis of cAMP and cGMP; SmPDE11 displayed considerable preferrence for cGMP over cAMP. These results and tools enable the pursuit of a rigorous drug discovery program based on inhibitors of S. mansoni PDEs. |
format |
Article in Journal/Newspaper |
author |
Jane C Munday Stefan Kunz Titilola D Kalejaiye Marco Siderius Susanne Schroeder Daniel Paape Ali H Alghamdi Zainab Abbasi Sheng Xiang Huang Anne-Marie Donachie Samia William Abdel Nasser Sabra Geert Jan Sterk Sanaa S Botros David G Brown Charles S Hoffman Rob Leurs Harry P de Koning |
author_facet |
Jane C Munday Stefan Kunz Titilola D Kalejaiye Marco Siderius Susanne Schroeder Daniel Paape Ali H Alghamdi Zainab Abbasi Sheng Xiang Huang Anne-Marie Donachie Samia William Abdel Nasser Sabra Geert Jan Sterk Sanaa S Botros David G Brown Charles S Hoffman Rob Leurs Harry P de Koning |
author_sort |
Jane C Munday |
title |
Cloning and functional complementation of ten Schistosoma mansoni phosphodiesterases expressed in the mammalian host stages. |
title_short |
Cloning and functional complementation of ten Schistosoma mansoni phosphodiesterases expressed in the mammalian host stages. |
title_full |
Cloning and functional complementation of ten Schistosoma mansoni phosphodiesterases expressed in the mammalian host stages. |
title_fullStr |
Cloning and functional complementation of ten Schistosoma mansoni phosphodiesterases expressed in the mammalian host stages. |
title_full_unstemmed |
Cloning and functional complementation of ten Schistosoma mansoni phosphodiesterases expressed in the mammalian host stages. |
title_sort |
cloning and functional complementation of ten schistosoma mansoni phosphodiesterases expressed in the mammalian host stages. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2020 |
url |
https://doi.org/10.1371/journal.pntd.0008447 https://doaj.org/article/dec44c4ba15a44afbe2fcc56f7a87eca |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 14, Iss 7, p e0008447 (2020) |
op_relation |
https://doi.org/10.1371/journal.pntd.0008447 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0008447 https://doaj.org/article/dec44c4ba15a44afbe2fcc56f7a87eca |
op_doi |
https://doi.org/10.1371/journal.pntd.0008447 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
14 |
container_issue |
7 |
container_start_page |
e0008447 |
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1766341898087170048 |