High content drug screening for Fanconi anemia therapeutics
Abstract Background Fanconi anemia is a rare disease clinically characterized by malformations, bone marrow failure and an increased risk of solid tumors and hematologic malignancies. The only therapies available are hematopoietic stem cell transplantation for bone marrow failure or leukemia, and su...
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| Format: | Article |
| Language: | English |
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BMC
2020-06-01
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| Series: | Orphanet Journal of Rare Diseases |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13023-020-01437-1 |
| _version_ | 1831678463624347648 |
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| author | Helena Montanuy Cristina Camps-Fajol Jordi Carreras-Puigvert Maria Häggblad Bo Lundgren Miriam Aza-Carmona Thomas Helleday Jordi Minguillón Jordi Surrallés |
| author_facet | Helena Montanuy Cristina Camps-Fajol Jordi Carreras-Puigvert Maria Häggblad Bo Lundgren Miriam Aza-Carmona Thomas Helleday Jordi Minguillón Jordi Surrallés |
| author_sort | Helena Montanuy |
| collection | DOAJ |
| description | Abstract Background Fanconi anemia is a rare disease clinically characterized by malformations, bone marrow failure and an increased risk of solid tumors and hematologic malignancies. The only therapies available are hematopoietic stem cell transplantation for bone marrow failure or leukemia, and surgical resection for solid tumors. Therefore, there is still an urgent need for new therapeutic options. With this aim, we developed a novel high-content cell-based screening assay to identify drugs with therapeutic potential in FA. Results A TALEN-mediated FANCA-deficient U2OS cell line was stably transfected with YFP-FANCD2 fusion protein. These cells were unable to form fluorescent foci or to monoubiquitinate endogenous or exogenous FANCD2 upon DNA damage and were more sensitive to mitomycin C when compared to the parental wild type counterpart. FANCA correction by retroviral infection restored the cell line’s ability to form FANCD2 foci and ubiquitinate FANCD2. The feasibility of this cell-based system was interrogated in a high content screening of 3802 compounds, including a Prestwick library of 1200 FDA-approved drugs. The potential hits identified were then individually tested for their ability to rescue FANCD2 foci and monoubiquitination, and chromosomal stability in the absence of FANCA. Conclusions While, unfortunately, none of the compounds tested were able to restore cellular FANCA-deficiency, our study shows the potential capacity to screen large compound libraries in the context of Fanconi anemia therapeutics in an optimized and cost-effective platform. |
| first_indexed | 2024-12-20T04:59:59Z |
| format | Article |
| id | doaj.art-64abf667360445cfaf72bee11ee1388b |
| institution | Directory Open Access Journal |
| issn | 1750-1172 |
| language | English |
| last_indexed | 2024-12-20T04:59:59Z |
| publishDate | 2020-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Orphanet Journal of Rare Diseases |
| spelling | doaj.art-64abf667360445cfaf72bee11ee1388b2022-12-21T19:52:35ZengBMCOrphanet Journal of Rare Diseases1750-11722020-06-011511910.1186/s13023-020-01437-1High content drug screening for Fanconi anemia therapeuticsHelena Montanuy0Cristina Camps-Fajol1Jordi Carreras-Puigvert2Maria Häggblad3Bo Lundgren4Miriam Aza-Carmona5Thomas Helleday6Jordi Minguillón7Jordi Surrallés8Department of Genetics and Microbiology, Universitat Autònoma de BarcelonaDepartment of Genetics and Microbiology, Universitat Autònoma de BarcelonaDepartment of Genetics and Microbiology, Universitat Autònoma de BarcelonaDivision of Translational Medicine and Chemical Biology, Science for Life Laboratory, Department of Molecular Biochemistry and Biophysics, Karolinska InstitutetDepartment of Biochemistry and Biophysics, SciLifelab, Stockholm UniversityInstitute of Medical and Molecular Genetics and Skeletal dysplasia multidisciplinary Unit, Hospital Universitario La Paz, Universidad Autónoma de MadridDivision of Translational Medicine and Chemical Biology, Science for Life Laboratory, Department of Molecular Biochemistry and Biophysics, Karolinska InstitutetDepartment of Genetics and Microbiology, Universitat Autònoma de BarcelonaDepartment of Genetics and Microbiology, Universitat Autònoma de BarcelonaAbstract Background Fanconi anemia is a rare disease clinically characterized by malformations, bone marrow failure and an increased risk of solid tumors and hematologic malignancies. The only therapies available are hematopoietic stem cell transplantation for bone marrow failure or leukemia, and surgical resection for solid tumors. Therefore, there is still an urgent need for new therapeutic options. With this aim, we developed a novel high-content cell-based screening assay to identify drugs with therapeutic potential in FA. Results A TALEN-mediated FANCA-deficient U2OS cell line was stably transfected with YFP-FANCD2 fusion protein. These cells were unable to form fluorescent foci or to monoubiquitinate endogenous or exogenous FANCD2 upon DNA damage and were more sensitive to mitomycin C when compared to the parental wild type counterpart. FANCA correction by retroviral infection restored the cell line’s ability to form FANCD2 foci and ubiquitinate FANCD2. The feasibility of this cell-based system was interrogated in a high content screening of 3802 compounds, including a Prestwick library of 1200 FDA-approved drugs. The potential hits identified were then individually tested for their ability to rescue FANCD2 foci and monoubiquitination, and chromosomal stability in the absence of FANCA. Conclusions While, unfortunately, none of the compounds tested were able to restore cellular FANCA-deficiency, our study shows the potential capacity to screen large compound libraries in the context of Fanconi anemia therapeutics in an optimized and cost-effective platform.http://link.springer.com/article/10.1186/s13023-020-01437-1Fanconi anemiaHigh content screeningDrug repositioningCell-based assay |
| spellingShingle | Helena Montanuy Cristina Camps-Fajol Jordi Carreras-Puigvert Maria Häggblad Bo Lundgren Miriam Aza-Carmona Thomas Helleday Jordi Minguillón Jordi Surrallés High content drug screening for Fanconi anemia therapeutics Orphanet Journal of Rare Diseases Fanconi anemia High content screening Drug repositioning Cell-based assay |
| title | High content drug screening for Fanconi anemia therapeutics |
| title_full | High content drug screening for Fanconi anemia therapeutics |
| title_fullStr | High content drug screening for Fanconi anemia therapeutics |
| title_full_unstemmed | High content drug screening for Fanconi anemia therapeutics |
| title_short | High content drug screening for Fanconi anemia therapeutics |
| title_sort | high content drug screening for fanconi anemia therapeutics |
| topic | Fanconi anemia High content screening Drug repositioning Cell-based assay |
| url | http://link.springer.com/article/10.1186/s13023-020-01437-1 |
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