A Phenotypic Screening Assay Identifies Modulators of Diamond Blackfan Anemia

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

Abstract

Diamond-Blackfan anemia (DBA) is a bone marrow failure syndrome caused by mutations in ribosomal protein genes. Pathogenic mechanisms are poorly understood but involve severely reduced proliferation of erythroid precursors. Because current DBA therapies are ineffective and associated with severe side effects, disease-specific therapies are urgently needed. We hypothesized that druggable molecular pathways underlying the defect can be revealed through phenotypic small-molecule screens. Accordingly, a screening assay was developed using c-kit+ fetal liver erythroid progenitors from a doxycycline-inducible DBA mouse model. The addition of doxycycline to the culture medium induces the phenotype and reduces proliferation to <10% of normal, such that rescue of proliferation can be used as a simple readout for screening. Here, we describe the assay rationale and efforts toward validation of a microtiter plate-compatible assay and its application in a pilot screen of 3871 annotated compounds. Ten hits demonstrated concentration-dependent activity, and we report a brief follow-up of one of these compounds. In conclusion, we established a robust scalable assay for screening molecules that rescue erythropoiesis in DBA.

Detaljer

Författare
Enheter & grupper
Externa organisationer
  • Karolinska Institute
  • University Medical Center Freiburg
  • AstraZeneca
Forskningsområden

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Cell- och molekylärbiologi

Nyckelord

Originalspråkengelska
Sidor (från-till)304-313
Antal sidor10
TidskriftSLAS discovery : advancing life sciences R & D
Volym24
Utgivningsnummer3
StatusPublished - 2019
PublikationskategoriForskning
Peer review utfördJa