Characterization of stem cell-derived liver and intestinal organoids as a model system to study nuclear receptor biology

https://doi.org/10.1016/j.bbadis.2016.12.004Get rights and content
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Highlights

  • NR mRNA expression and function is similar between organoids and respective tissues.

  • Fxrα, Lxrα/β, Pparα/γ activation showed induction of multiple target genes.

  • NR target genes were induced in WT, but not in NR −/− organoids.

  • Organoids are easy to use and suitable to study NR biology and targetability.

  • Trpm6, Slc26a3, Ang1, and Rnase4 are novel target genes of Fxr in the intestine.

Abstract

Nuclear receptors (NRs) are ligand-activated transcription factors regulating a large variety of processes involved in reproduction, development, and metabolism. NRs are ideal drug targets because they are activated by lipophilic ligands that easily pass cell membranes. Immortalized cell lines recapitulate NR biology poorly and generating primary cultures is laborious and requires a constant need for donor material. There is a clear need for development of novel preclinical model systems that better resemble human physiology. Uncertainty due to technical limitations early in drug development is often the cause of preclinical drugs not reaching the clinic. Here, we studied whether organoids, mini-organs derived from the respective mouse tissue's stem cells, can serve as a novel model system to study NR biology and targetability. We characterized mRNA expression profiles of the NR superfamily in mouse liver, ileum, and colon organoids. Tissue-specific expression patterns were largely maintained in the organoids, indicating their suitability for NR research. Metabolic NRs Fxrα, Lxrα, Lxrβ, Pparα, and Pparγ induced expression of and binding to endogenous target genes. Transcriptome analyses of wildtype colon organoids stimulated with Rosiglitazone showed that lipid metabolism was the highest significant changed function, greatly mimicking the PPARs and Rosiglitazone function in vivo. Finally, using organoids we identify Trpm6, Slc26a3, Ang1, and Rnase4, as novel Fxr target genes. Our results demonstrate that organoids represent a framework to study NR biology that can be further expanded to human organoids to improve preclinical testing of novel drugs that target this pharmacologically important class of ligand activated transcription factors.

Abbreviations

ChIP
chromatin immunoprecipitation
DIC
differential interference contrast
DM
differentiation medium
DSG
Di(N-succinimidyl) glutarate
EM
expansion medium
Fxr
farnesoid x receptor
IPA
Ingenuity Pathway Analysis
Lxr
Liver x receptor
NR
nuclear receptor
OCA
obeticholic acid
Ppar
Peroxisome proliferator-activated receptor
RMA
robust multiarray analysis
Rosi
Rosiglitazone

Keywords

Nuclear receptors
Fxr
Lxr
Ppar
Ligands
Organoids

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