2. Academic Validation
  3. High content drug screening for Fanconi anemia therapeutics

High content drug screening for Fanconi anemia therapeutics

  • Orphanet J Rare Dis. 2020 Jun 30;15(1):170. doi: 10.1186/s13023-020-01437-1.
Helena Montanuy 1,  Cristina Camps-Fajol 1 2,  Jordi Carreras-Puigvert 1 2 3,  Maria Häggblad 4 5,  Bo Lundgren 6,  Miriam Aza-Carmona 7 8,  Thomas Helleday 4,  Jordi Minguillón 1 3 9,  Jordi Surrallés 10 11 12 13
Affiliations

Affiliations

  • 1 Department of Genetics and Microbiology, Universitat Autònoma de Barcelona, Barcelona, Spain.
  • 2 Join Research Unit on Genomic Medicine UAB-Sant Pau, Biomedical Research Institute, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain.
  • 3 Centro de Investigación Biomédica en Red de Enfermedades raras, Barcelona, Spain.
  • 4 Division of Translational Medicine and Chemical Biology, Science for Life Laboratory, Department of Molecular Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
  • 5 Currently at Division of Genome Biology, Science for Life Laboratory, Department of Molecular Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
  • 6 Department of Biochemistry and Biophysics, SciLifelab, Stockholm University, Stockholm, SE, Sweden.
  • 7 Institute of Medical and Molecular Genetics and Skeletal dysplasia multidisciplinary Unit, Hospital Universitario La Paz, Universidad Autónoma de Madrid, IdiPaz, Madrid, Spain.
  • 8 Centro de Investigación Biomédica en Red de Enfermedades Raras, Madrid, Spain.
  • 9 Genetics Department, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain.
  • 10 Department of Genetics and Microbiology, Universitat Autònoma de Barcelona, Barcelona, Spain. Jordi.surralles@uab.cat.
  • 11 Join Research Unit on Genomic Medicine UAB-Sant Pau, Biomedical Research Institute, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain. Jordi.surralles@uab.cat.
  • 12 Centro de Investigación Biomédica en Red de Enfermedades raras, Barcelona, Spain. Jordi.surralles@uab.cat.
  • 13 Genetics Department, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain. Jordi.surralles@uab.cat.
PMID: 32605631 DOI: 10.1186/s13023-020-01437-1
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.

Keywords

Cell-based assay; Drug repositioning; Fanconi anemia; High content screening.

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