High-content screening identifies small molecules that remove nuclear foci, affect MBNL distribution and CELF1 protein levels via a PKC-independent pathway in myotonic dystrophy cell lines
Myotonic dystrophy (DM) is a multi-system neuromuscular disorder for which there is no treatment. We have developed a medium throughput phenotypic assay, based on the identification of nuclear foci in DM patient cell lines using in situ hybridization and high-content imaging to screen for potentiall...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
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Oxford University Press
2014
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| Online Access: | https://eprints.nottingham.ac.uk/2905/ |
| _version_ | 1848790905148932096 |
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| author | Ketley, Ami Chen, Catherine Z. Li, Xin Arya, Sukrat Robinson, Thelma E. Granados-Riveron, Javier T. Udosen, Inyang Morris, Glenn E. Holt, Ian Furling, Dennis Chaouch, Soraya Haworth, Ben Southall, Noel Shinn, Paul Zheng, Wei Austin, Christopher P. Hayes, Christopher J. Brook, J. David |
| author_facet | Ketley, Ami Chen, Catherine Z. Li, Xin Arya, Sukrat Robinson, Thelma E. Granados-Riveron, Javier T. Udosen, Inyang Morris, Glenn E. Holt, Ian Furling, Dennis Chaouch, Soraya Haworth, Ben Southall, Noel Shinn, Paul Zheng, Wei Austin, Christopher P. Hayes, Christopher J. Brook, J. David |
| author_sort | Ketley, Ami |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Myotonic dystrophy (DM) is a multi-system neuromuscular disorder for which there is no treatment. We have developed a medium throughput phenotypic assay, based on the identification of nuclear foci in DM patient cell lines using in situ hybridization and high-content imaging to screen for potentially useful therapeutic compounds. A series of further assays based on molecular features of DM have also been employed. Two compounds that reduce and/or remove nuclear foci have been identified, Ro 31-8220 and chromomycin A3. Ro 31-8220 is a PKC inhibitor, previously shown to affect the hyperphosphorylation of CELF1 and ameliorate the cardiac phenotype in a DM1 mouse model. We show that the same compound eliminates nuclear foci, reduces MBNL1 protein in the nucleus, affects ATP2A1 alternative splicing and reduces steady-state levels of CELF1 protein. We demonstrate that this effect is independent of PKC activity and conclude that this compound may be acting on alternative kinase targets within DM pathophysiology. Understanding the activity profile for this compound is key for the development of targeted therapeutics in the treatment of DM. |
| first_indexed | 2025-11-14T18:20:02Z |
| format | Article |
| id | nottingham-2905 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T18:20:02Z |
| publishDate | 2014 |
| publisher | Oxford University Press |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-29052020-05-04T16:45:18Z https://eprints.nottingham.ac.uk/2905/ High-content screening identifies small molecules that remove nuclear foci, affect MBNL distribution and CELF1 protein levels via a PKC-independent pathway in myotonic dystrophy cell lines Ketley, Ami Chen, Catherine Z. Li, Xin Arya, Sukrat Robinson, Thelma E. Granados-Riveron, Javier T. Udosen, Inyang Morris, Glenn E. Holt, Ian Furling, Dennis Chaouch, Soraya Haworth, Ben Southall, Noel Shinn, Paul Zheng, Wei Austin, Christopher P. Hayes, Christopher J. Brook, J. David Myotonic dystrophy (DM) is a multi-system neuromuscular disorder for which there is no treatment. We have developed a medium throughput phenotypic assay, based on the identification of nuclear foci in DM patient cell lines using in situ hybridization and high-content imaging to screen for potentially useful therapeutic compounds. A series of further assays based on molecular features of DM have also been employed. Two compounds that reduce and/or remove nuclear foci have been identified, Ro 31-8220 and chromomycin A3. Ro 31-8220 is a PKC inhibitor, previously shown to affect the hyperphosphorylation of CELF1 and ameliorate the cardiac phenotype in a DM1 mouse model. We show that the same compound eliminates nuclear foci, reduces MBNL1 protein in the nucleus, affects ATP2A1 alternative splicing and reduces steady-state levels of CELF1 protein. We demonstrate that this effect is independent of PKC activity and conclude that this compound may be acting on alternative kinase targets within DM pathophysiology. Understanding the activity profile for this compound is key for the development of targeted therapeutics in the treatment of DM. Oxford University Press 2014-03-06 Article PeerReviewed Ketley, Ami, Chen, Catherine Z., Li, Xin, Arya, Sukrat, Robinson, Thelma E., Granados-Riveron, Javier T., Udosen, Inyang, Morris, Glenn E., Holt, Ian, Furling, Dennis, Chaouch, Soraya, Haworth, Ben, Southall, Noel, Shinn, Paul, Zheng, Wei, Austin, Christopher P., Hayes, Christopher J. and Brook, J. David (2014) High-content screening identifies small molecules that remove nuclear foci, affect MBNL distribution and CELF1 protein levels via a PKC-independent pathway in myotonic dystrophy cell lines. Human Molecular Genetics, 23 (6). pp. 1551-1562. ISSN 1460-2083 http://hmg.oxfordjournals.org/content/23/6/1551 doi:10.1093/hmg/ddt542 doi:10.1093/hmg/ddt542 |
| spellingShingle | Ketley, Ami Chen, Catherine Z. Li, Xin Arya, Sukrat Robinson, Thelma E. Granados-Riveron, Javier T. Udosen, Inyang Morris, Glenn E. Holt, Ian Furling, Dennis Chaouch, Soraya Haworth, Ben Southall, Noel Shinn, Paul Zheng, Wei Austin, Christopher P. Hayes, Christopher J. Brook, J. David High-content screening identifies small molecules that remove nuclear foci, affect MBNL distribution and CELF1 protein levels via a PKC-independent pathway in myotonic dystrophy cell lines |
| title | High-content screening identifies small molecules that remove nuclear foci, affect MBNL distribution and CELF1 protein levels via a PKC-independent pathway in myotonic dystrophy cell lines |
| title_full | High-content screening identifies small molecules that remove nuclear foci, affect MBNL distribution and CELF1 protein levels via a PKC-independent pathway in myotonic dystrophy cell lines |
| title_fullStr | High-content screening identifies small molecules that remove nuclear foci, affect MBNL distribution and CELF1 protein levels via a PKC-independent pathway in myotonic dystrophy cell lines |
| title_full_unstemmed | High-content screening identifies small molecules that remove nuclear foci, affect MBNL distribution and CELF1 protein levels via a PKC-independent pathway in myotonic dystrophy cell lines |
| title_short | High-content screening identifies small molecules that remove nuclear foci, affect MBNL distribution and CELF1 protein levels via a PKC-independent pathway in myotonic dystrophy cell lines |
| title_sort | high-content screening identifies small molecules that remove nuclear foci, affect mbnl distribution and celf1 protein levels via a pkc-independent pathway in myotonic dystrophy cell lines |
| url | https://eprints.nottingham.ac.uk/2905/ https://eprints.nottingham.ac.uk/2905/ https://eprints.nottingham.ac.uk/2905/ |