High content drug screening for Fanconi anemia therapeutics
Montanuy, Helena (Universitat Autònoma de Barcelona. Departament de Genètica i de Microbiologia)
Camps-Fajol, Cristina (Institut d'Investigació Biomèdica Sant Pau)
Carreras-Puigvert, Jordi (Centro de Investigación Biomédica en Red de Enfermedades Raras)
Häggblad, Maria (Karolinska Institutet (Estocolm, Suècia))
Lundgren, Bo (Stockholm University)
Aza-Carmona, Miriam (Centro de Investigación Biomédica en Red de Enfermedades Raras)
Helleday, Thomas (Karolinska Institutet (Estocolm, Suècia). Department of Molecular Biochemistry and Biophysics)
Minguillón Pedreño, Jordi (Institut d'Investigació Biomèdica Sant Pau)
Surrallés i Calonge, Jordi (Institut d'Investigació Biomèdica Sant Pau)
Universitat Autònoma de Barcelona

Date:	2020
Abstract:	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. 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. 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.
Grants:	Ministerio de Economía, Industria y Competitividad SAF2015-64152-R Ministerio de Economía, Industria y Competitividad CB06/07/0023
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Fanconi anemia ; High content screening ; Drug repositioning ; Cell-based assay
Published in:	Orphanet Journal of Rare Diseases, Vol. 15 (june 2020) , ISSN 1750-1172

DOI: 10.1186/s13023-020-01437-1
PMID: 32605631

9 p, 1.5 MB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Recerca Sant Pau
Articles > Research articles
Articles > Published articles