A chemical screen identifies small molecules that regulate hepcidin expression
STAT3 Transcription Factor
0301 basic medicine
Cell Survival
Recombinant Fusion Proteins
Smad Proteins
Transfection
Small Molecule Libraries
03 medical and health sciences
Hepcidins
Genes, Reporter
Drug Discovery
Humans
Luciferases
Promoter Regions, Genetic
Molecular Biology
0303 health sciences
Stat3
Interleukin-6
Cell Biology
Hematology
Hep G2 Cells
High-Throughput Screening Assays
3. Good health
Gene Expression Regulation
Bone Morphogenetic Proteins
Thalassemia
Molecular Medicine
Hemochromatosis
Bone morphogenic protein
Databases, Chemical
Plasmids
Signal Transduction
DOI:
10.1016/j.bcmd.2014.06.002
Publication Date:
2014-07-04T03:45:32Z
AUTHORS (7)
ABSTRACT
Hepcidin, a peptide hormone produced in the liver, decreases intestinal iron absorption and macrophage iron release via effects on ferroportin. Bone morphogenic protein and Stat3 signaling regulate Hepcidin's transcription. Hepcidin is a potential drug target for patients with iron overload syndromes because its levels are inappropriately low in these individuals. To generate a tool for identifying small molecules that modulate Hepcidin expression, we stably transfected human hepatocytes (HepG2) cells with a reporter construct containing 2.7kb of the human Hepcidin promoter upstream of a firefly reporter gene. We used high throughput methods to screen 10,169 chemicals in duplicate for their effect on Hepcidin expression and cell viability. Regulators were identified as chemicals that caused a change >3 standard deviations above or >1 standard deviation below the mean of the other chemicals (z-score >3 or <1), while not adversely affecting cell viability, quantified by fluorescence assay. Following validation assays, we identified 16 chemicals in a broad range of functional classes that promote Hepcidin expression. All of the chemicals identified increased expression of bone morphogenic protein-dependent and/or Stat3-dependent genes, however none of them strongly increased phosphorylation of Smad1,5,8 or Stat3.
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CITATIONS (17)
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