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Use of radiolabelled choline as a pharmacodynamic marker for the signal transduction inhibitor geldanamycin

There is an urgent need to develop non-invasive pharmacodynamic endpoints for the evaluation of new molecular therapeutics that inhibit signal transduction. We hypothesised that, when labelled appropriately, changes in choline kinetics could be used to assess geldanamycin pharmacodynamics, which inv...

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Autores principales: Liu, D, Hutchinson, O C, Osman, S, Price, P, Workman, P, Aboagye, E O
Formato: Texto
Lenguaje:English
Publicado: Nature Publishing Group 2002
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2364261/
https://www.ncbi.nlm.nih.gov/pubmed/12232764
http://dx.doi.org/10.1038/sj.bjc.6600558
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author Liu, D
Hutchinson, O C
Osman, S
Price, P
Workman, P
Aboagye, E O
author_facet Liu, D
Hutchinson, O C
Osman, S
Price, P
Workman, P
Aboagye, E O
author_sort Liu, D
collection PubMed
description There is an urgent need to develop non-invasive pharmacodynamic endpoints for the evaluation of new molecular therapeutics that inhibit signal transduction. We hypothesised that, when labelled appropriately, changes in choline kinetics could be used to assess geldanamycin pharmacodynamics, which involves inhibition of the HSP90 molecular chaperone→Raf1→Mitogenic Extracellular Kinase→Extracellular Signal-Regulated Kinase 1 and 2 signal transduction pathway. Towards identifying a potential pharmacodynamic marker response, we have studied radiolabelled choline metabolism in HT29 human colon carcinoma cells following treatment with geldanamycin. We studied the effects of geldanamycin, on net cellular accumulation of (methyl-(14)C)choline and (methyl-(14)C)phosphocholine production. In parallel experiments, the effects of geldanamycin on extracellular signal-regulated kinase 1 and 2 phosphorylation and cell viability were also assessed. Additional validation studies were carried out with the mitogenic extracellular kinase inhibitor U0126 as a positive control; a cyclin-dependent kinase-2 inhibitor roscovitine and the phosphatidylinositol 3-kinase inhibitor LY294002 as negative controls. Hemicholinium-3, an inhibitor of choline transport and choline kinase activity was included as an additional control. In exponentially growing HT29 cells, geldanamycin inhibited extracellular signal-regulated kinase 1 and 2 phosphorylation in a concentration- and time-dependent manner. These changes were associated with a reduction in (methyl-(14)C)choline uptake, (methyl-(14)C) phosphocholine production and cell viability. Brief exposure to U0126, suppressed phosphocholine production to the same extent as Hemicholinium-3. In contrast to geldanamycin and U0126, which act upstream of extracellular signal-regulated kinase 1 and 2, roscovitine and LY294002 failed to suppress phosphocholine production. Our results suggest that when labelled with carbon-11 isotope, (methyl-(11)C)choline may be a useful pharmacodynamic marker for the non-invasive evaluation of geldanamycin analogues. British Journal of Cancer (2002) 87, 783–789. doi:10.1038/sj.bjc.6600558 www.bjcancer.com © 2002 Cancer Research UK
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spelling pubmed-23642612009-09-10 Use of radiolabelled choline as a pharmacodynamic marker for the signal transduction inhibitor geldanamycin Liu, D Hutchinson, O C Osman, S Price, P Workman, P Aboagye, E O Br J Cancer Experimental Therapeutics There is an urgent need to develop non-invasive pharmacodynamic endpoints for the evaluation of new molecular therapeutics that inhibit signal transduction. We hypothesised that, when labelled appropriately, changes in choline kinetics could be used to assess geldanamycin pharmacodynamics, which involves inhibition of the HSP90 molecular chaperone→Raf1→Mitogenic Extracellular Kinase→Extracellular Signal-Regulated Kinase 1 and 2 signal transduction pathway. Towards identifying a potential pharmacodynamic marker response, we have studied radiolabelled choline metabolism in HT29 human colon carcinoma cells following treatment with geldanamycin. We studied the effects of geldanamycin, on net cellular accumulation of (methyl-(14)C)choline and (methyl-(14)C)phosphocholine production. In parallel experiments, the effects of geldanamycin on extracellular signal-regulated kinase 1 and 2 phosphorylation and cell viability were also assessed. Additional validation studies were carried out with the mitogenic extracellular kinase inhibitor U0126 as a positive control; a cyclin-dependent kinase-2 inhibitor roscovitine and the phosphatidylinositol 3-kinase inhibitor LY294002 as negative controls. Hemicholinium-3, an inhibitor of choline transport and choline kinase activity was included as an additional control. In exponentially growing HT29 cells, geldanamycin inhibited extracellular signal-regulated kinase 1 and 2 phosphorylation in a concentration- and time-dependent manner. These changes were associated with a reduction in (methyl-(14)C)choline uptake, (methyl-(14)C) phosphocholine production and cell viability. Brief exposure to U0126, suppressed phosphocholine production to the same extent as Hemicholinium-3. In contrast to geldanamycin and U0126, which act upstream of extracellular signal-regulated kinase 1 and 2, roscovitine and LY294002 failed to suppress phosphocholine production. Our results suggest that when labelled with carbon-11 isotope, (methyl-(11)C)choline may be a useful pharmacodynamic marker for the non-invasive evaluation of geldanamycin analogues. British Journal of Cancer (2002) 87, 783–789. doi:10.1038/sj.bjc.6600558 www.bjcancer.com © 2002 Cancer Research UK Nature Publishing Group 2002-09-23 2002-09-23 /pmc/articles/PMC2364261/ /pubmed/12232764 http://dx.doi.org/10.1038/sj.bjc.6600558 Text en Copyright © 2002 Cancer Research UK https://creativecommons.org/licenses/by/4.0/This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material.If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.
spellingShingle Experimental Therapeutics
Liu, D
Hutchinson, O C
Osman, S
Price, P
Workman, P
Aboagye, E O
Use of radiolabelled choline as a pharmacodynamic marker for the signal transduction inhibitor geldanamycin
title Use of radiolabelled choline as a pharmacodynamic marker for the signal transduction inhibitor geldanamycin
title_full Use of radiolabelled choline as a pharmacodynamic marker for the signal transduction inhibitor geldanamycin
title_fullStr Use of radiolabelled choline as a pharmacodynamic marker for the signal transduction inhibitor geldanamycin
title_full_unstemmed Use of radiolabelled choline as a pharmacodynamic marker for the signal transduction inhibitor geldanamycin
title_short Use of radiolabelled choline as a pharmacodynamic marker for the signal transduction inhibitor geldanamycin
title_sort use of radiolabelled choline as a pharmacodynamic marker for the signal transduction inhibitor geldanamycin
topic Experimental Therapeutics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2364261/
https://www.ncbi.nlm.nih.gov/pubmed/12232764
http://dx.doi.org/10.1038/sj.bjc.6600558
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