Phenotypic, chemical and functional characterization of cyclic nucleotide phosphodiesterase 4 (PDE4) as a potential anthelmintic drug target
| العنوان: | Phenotypic, chemical and functional characterization of cyclic nucleotide phosphodiesterase 4 (PDE4) as a potential anthelmintic drug target |
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| المؤلفون: | Maliwan Meewan, Lin Lin, Fernando Rock, Kurt Jarnagin, Thavy Long, Katherine A. Cunningham, Da Shi, James H. McKerrow, George A. Lemieux, Alberto A. Rascón, Liliana Rojo-Arreola, Larissa M. Podust, Nelly El-Sakkary, Ruben Abagyan, Kaveh Ashrafi, Conor R. Caffrey |
| المساهمون: | Geary, Timothy G |
| المصدر: | PLoS Neglected Tropical Diseases PLoS neglected tropical diseases, vol 11, iss 7 Long, T; Rojo-Arreola, L; Shi, D; El-Sakkary, N; Jarnagin, K; Rock, F; et al.(2017). Phenotypic, chemical and functional characterization of cyclic nucleotide phosphodiesterase 4 (PDE4) as a potential anthelmintic drug target. PLoS Neglected Tropical Diseases, 11(7). doi: 10.1371/journal.pntd.0005680. UC San Francisco: Retrieved from: http://www.escholarship.org/uc/item/37t2q5gm PLoS Neglected Tropical Diseases, Vol 11, Iss 7, p e0005680 (2017) |
| بيانات النشر: | Public Library of Science, 2017. |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | 0301 basic medicine, Schistosoma Mansoni, Nematoda, Mutant, Medical and Health Sciences, Biochemistry, Animals, Genetically Modified, Database and Informatics Methods, 0302 clinical medicine, Catalytic Domain, 2.1 Biological and endogenous factors, Aetiology, Enzyme Inhibitors, Cloning, Molecular, Schistosoma Japonicum, Caenorhabditis elegans, Genetics, Anthelmintics, biology, Drug discovery, lcsh:Public aspects of medicine, Schistosoma japonicum, Schistosoma mansoni, Animal Models, Biological Sciences, 3. Good health, Infectious Diseases, Experimental Organism Systems, 5.1 Pharmaceuticals, 030220 oncology & carcinogenesis, Schistosoma, Development of treatments and therapeutic interventions, Type 4, Sequence Analysis, Locomotion, Biotechnology, Research Article, Cyclic Nucleotide Phosphodiesterases, lcsh:Arctic medicine. Tropical medicine, lcsh:RC955-962, Bioinformatics, Transgene, Phenotypic screening, Genetically Modified, Research and Analysis Methods, 03 medical and health sciences, Rare Diseases, Model Organisms, Sequence Motif Analysis, Tropical Medicine, Helminths, Animals, Public Health, Environmental and Occupational Health, Wild type, Organisms, Molecular, Biology and Life Sciences, lcsh:RA1-1270, biology.organism_classification, Molecular biology, Invertebrates, Schistosoma Haematobium, Cyclic Nucleotide Phosphodiesterases, Type 4, Vector-Borne Diseases, 030104 developmental biology, Orphan Drug, Good Health and Well Being, Caenorhabditis, Enzymology, Phosphodiesterase 4 Inhibitors, Sequence Alignment, Cloning |
| الوصف: | Background Reliance on just one drug to treat the prevalent tropical disease, schistosomiasis, spurs the search for new drugs and drug targets. Inhibitors of human cyclic nucleotide phosphodiesterases (huPDEs), including PDE4, are under development as novel drugs to treat a range of chronic indications including asthma, chronic obstructive pulmonary disease and Alzheimer’s disease. One class of huPDE4 inhibitors that has yielded marketed drugs is the benzoxaboroles (Anacor Pharmaceuticals). Methodology/Principal findings A phenotypic screen involving Schistosoma mansoni and 1,085 benzoxaboroles identified a subset of huPDE4 inhibitors that induced parasite hypermotility and degeneration. To uncover the putative schistosome PDE4 target, we characterized four PDE4 sequences (SmPDE4A-D) in the parasite’s genome and transcriptome, and cloned and recombinantly expressed the catalytic domain of SmPDE4A. Among a set of benzoxaboroles and catechol inhibitors that differentially inhibit huPDE4, a relationship between the inhibition of SmPDE4A, and parasite hypermotility and degeneration, was measured. To validate SmPDE4A as the benzoxaborole molecular target, we first generated Caenorhabditis elegans lines that express a cDNA for smpde4a on a pde4(ce268) mutant (hypermotile) background: the smpde4a transgene restored mutant worm motility to that of the wild type. We then showed that benzoxaborole inhibitors of SmPDE4A that induce hypermotility in the schistosome also elicit a hypermotile response in the C. elegans lines that express the smpde4a transgene, thereby confirming SmPDE4A as the relevant target. Conclusions/Significance The orthogonal chemical, biological and genetic strategies employed identify SmPDE4A’s contribution to parasite motility and degeneration, and its potential as a drug target. Transgenic C. elegans is highlighted as a potential screening tool to optimize small molecule chemistries to flatworm molecular drug targets. Author summary Just one drug, praziquantel (PZQ), is available to treat schistosomiasis, a flatworm disease that infects over 240 million people, mainly in Africa. With the expanding distribution of PZQ, and the associated threat of drug resistance, new drugs and drug targets are needed. We screened Schistosoma mansoni worms with over 1,000 benzoxaborole chemical molecules from Anacor Pharmaceuticals to identify a subset of human cyclic nucleotide phosphodiesterase 4 (huPDE4) inhibitors that cause parasite hypermotility and degeneration. We identified four PDE4 genes in the genome of the parasite and recombinantly expressed one of them (SmPDE4A) in bacteria. This enzyme was then used to uncover a relationship between the degree of enzyme inhibition, and the generation of parasite hypermotility and degeneration. To understand whether the SmPDE4A enzyme is the target of the benzoxaboroles in the parasite, we incorporated the coding DNA for SmPDE4A into the model nematode Caenorhabditis elegans that lacked its own functional PDE4 and, as a consequence, was hypermotile. These ‘transgenic’ worms displayed normal motility which could be increased by applying the most potent huPDE4 benzoxaborole inhibitors. In summary, the chemical, biological and genetic approaches taken identify SmPDE4A as a potential drug target for treating schistosomiasis. The potential value of C. elegans as a tool to test and optimize therapeutic chemistries for a flatworm disease is also highlighted. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 1935-2735 1935-2727 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::aa394f24515da5f88d2c848a0a1003bb http://europepmc.org/articles/PMC5526615 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....aa394f24515da5f88d2c848a0a1003bb |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19352735 19352727 |
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