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Limitations in a frataxin knockdown cell model for Friedreich ataxia in a high-throughput drug screen
BACKGROUND: Pharmacological high-throughput screening (HTS) represents a powerful strategy for drug discovery in genetic diseases, particularly when the full spectrum of pathological dysfunctions remains unclear, such as in Friedreich ataxia (FRDA). FRDA, the most common recessive ataxia, results fr...
Autores principales: | , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2009
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2744904/ https://www.ncbi.nlm.nih.gov/pubmed/19703283 http://dx.doi.org/10.1186/1471-2377-9-46 |
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author | Calmels, Nadège Seznec, Hervé Villa, Pascal Reutenauer, Laurence Hibert, Marcel Haiech, Jacques Rustin, Pierre Koenig, Michel Puccio, Hélène |
author_facet | Calmels, Nadège Seznec, Hervé Villa, Pascal Reutenauer, Laurence Hibert, Marcel Haiech, Jacques Rustin, Pierre Koenig, Michel Puccio, Hélène |
author_sort | Calmels, Nadège |
collection | PubMed |
description | BACKGROUND: Pharmacological high-throughput screening (HTS) represents a powerful strategy for drug discovery in genetic diseases, particularly when the full spectrum of pathological dysfunctions remains unclear, such as in Friedreich ataxia (FRDA). FRDA, the most common recessive ataxia, results from a generalized deficiency of mitochondrial and cytosolic iron-sulfur cluster (ISC) proteins activity, due to a partial loss of frataxin function, a mitochondrial protein proposed to function as an iron-chaperone for ISC biosynthesis. In the absence of measurable catalytic function for frataxin, a cell-based assay is required for HTS assay. METHODS: Using a targeted ribozyme strategy in murine fibroblasts, we have developed a cellular model with strongly reduced levels of frataxin. We have used this model to screen the Prestwick Chemical Library, a collection of one thousand off-patent drugs, for potential molecules for FRDA. RESULTS: The frataxin deficient cell lines exhibit a proliferation defect, associated with an ISC enzyme deficit. Using the growth defect as end-point criteria, we screened the Prestwick Chemical Library. However no molecule presented a significant and reproducible effect on the proliferation rate of frataxin deficient cells. Moreover over numerous passages, the antisense ribozyme fibroblast cell lines revealed an increase in frataxin residual level associated with the normalization of ISC enzyme activities. However, the ribozyme cell lines and FRDA patient cells presented an increase in Mthfd2 transcript, a mitochondrial enzyme that was previously shown to be upregulated at very early stages of the pathogenesis in the cardiac mouse model. CONCLUSION: Although no active hit has been identified, the present study demonstrates the feasibility of using a cell-based approach to HTS for FRDA. Furthermore, it highlights the difficulty in the development of a stable frataxin-deficient cell model, an essential condition for productive HTS in the future. |
format | Text |
id | pubmed-2744904 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-27449042009-09-16 Limitations in a frataxin knockdown cell model for Friedreich ataxia in a high-throughput drug screen Calmels, Nadège Seznec, Hervé Villa, Pascal Reutenauer, Laurence Hibert, Marcel Haiech, Jacques Rustin, Pierre Koenig, Michel Puccio, Hélène BMC Neurol Research Article BACKGROUND: Pharmacological high-throughput screening (HTS) represents a powerful strategy for drug discovery in genetic diseases, particularly when the full spectrum of pathological dysfunctions remains unclear, such as in Friedreich ataxia (FRDA). FRDA, the most common recessive ataxia, results from a generalized deficiency of mitochondrial and cytosolic iron-sulfur cluster (ISC) proteins activity, due to a partial loss of frataxin function, a mitochondrial protein proposed to function as an iron-chaperone for ISC biosynthesis. In the absence of measurable catalytic function for frataxin, a cell-based assay is required for HTS assay. METHODS: Using a targeted ribozyme strategy in murine fibroblasts, we have developed a cellular model with strongly reduced levels of frataxin. We have used this model to screen the Prestwick Chemical Library, a collection of one thousand off-patent drugs, for potential molecules for FRDA. RESULTS: The frataxin deficient cell lines exhibit a proliferation defect, associated with an ISC enzyme deficit. Using the growth defect as end-point criteria, we screened the Prestwick Chemical Library. However no molecule presented a significant and reproducible effect on the proliferation rate of frataxin deficient cells. Moreover over numerous passages, the antisense ribozyme fibroblast cell lines revealed an increase in frataxin residual level associated with the normalization of ISC enzyme activities. However, the ribozyme cell lines and FRDA patient cells presented an increase in Mthfd2 transcript, a mitochondrial enzyme that was previously shown to be upregulated at very early stages of the pathogenesis in the cardiac mouse model. CONCLUSION: Although no active hit has been identified, the present study demonstrates the feasibility of using a cell-based approach to HTS for FRDA. Furthermore, it highlights the difficulty in the development of a stable frataxin-deficient cell model, an essential condition for productive HTS in the future. BioMed Central 2009-08-24 /pmc/articles/PMC2744904/ /pubmed/19703283 http://dx.doi.org/10.1186/1471-2377-9-46 Text en Copyright © 2009 Calmels et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Calmels, Nadège Seznec, Hervé Villa, Pascal Reutenauer, Laurence Hibert, Marcel Haiech, Jacques Rustin, Pierre Koenig, Michel Puccio, Hélène Limitations in a frataxin knockdown cell model for Friedreich ataxia in a high-throughput drug screen |
title | Limitations in a frataxin knockdown cell model for Friedreich ataxia in a high-throughput drug screen |
title_full | Limitations in a frataxin knockdown cell model for Friedreich ataxia in a high-throughput drug screen |
title_fullStr | Limitations in a frataxin knockdown cell model for Friedreich ataxia in a high-throughput drug screen |
title_full_unstemmed | Limitations in a frataxin knockdown cell model for Friedreich ataxia in a high-throughput drug screen |
title_short | Limitations in a frataxin knockdown cell model for Friedreich ataxia in a high-throughput drug screen |
title_sort | limitations in a frataxin knockdown cell model for friedreich ataxia in a high-throughput drug screen |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2744904/ https://www.ncbi.nlm.nih.gov/pubmed/19703283 http://dx.doi.org/10.1186/1471-2377-9-46 |
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