T0901317 is a dual LXR/FXR agonist

doi:10.1016/j.ymgme.2004.07.007

Molecular Genetics and Metabolism

Volume 83, Issues 1–2, September–October 2004, Pages 184-187

https://doi.org/10.1016/j.ymgme.2004.07.007 Get rights and content

Abstract

We characterize the ability of the liver X receptor (LXRα [NR1H3] and LXRβ [NR1H2]) agonist, T0901317, to activate the farnesoid X receptor (FXR [NR4H4]). Although T0901317 is a much more potent activator of LXR than FXR, this ligand actually activates FXR more potently than a natural bile acid FXR ligand, chenodeoxycholic acid. Thus, the FXR activity of T0901317 must be considered when utilizing this agonist as a pharmacological tool to investigate LXR function.

Introduction

The liver X receptors (LXRα [NR1H3] and LXRβ [NR1H2]), members of the nuclear receptor superfamily, function as receptors for oxidized cholesterol and regulate a variety of physiological processes including cholesterol metabolism and transport, lipogenesis, gluconeogenesis, and inflammation. Synthetic LXR agonists have been proposed to have potential utility in treatment of disorders such as dyslipidemia, atherosclerosis, and diabetes [1]. To this end, high affinity LXR agonists, such as T0901317 and GW3965, have been described, which have allowed for the identification and characterization of many physiological processes regulated by LXR [2], [3].

Since T0901317 is often used as a specific LXR agonist “tool” to define the physiological effects of this receptor both in vitro and in vivo, we tested this ligand for activity against related receptors. Using cell-based transfection and biochemical ligand sensing assays, we found that T0901317 also acts as an FXR agonist. Like LXR, the farnesoid X receptor (FXR [NR1H4]) was originally identified as an orphan member of the nuclear receptor superfamily [4], [5]. FXR was later identified as the physiological receptor for bile acids and shown to regulate a feedback loop for bile acid transport and synthesis as well as modulating additional functions in lipid metabolism [6].

Section snippets

Cell culture and transfections

A Gal4 DNA-binding domain–human FXR ligand-binding domain fusion (Gal4DBD-FXRLBD) transfection assay was utilized as previously described with several modifications [7]. HEK293 cells were cultured in 3:1 DMEM:F-12 containing 10% fetal bovine serum and supplemented with 1% penicillin and streptomycin, 1% l-glutamine, and 20 mM Hepes. Forty-eight hours before transfection, cells were seeded at 6 × 10⁶ cells/T225 flask in 30 ml growth media. Cells were transfected with Fugene transfection reagent

Results

As shown in Fig. 1A, HEK293 cells transfected with a Gal4 DNA-binding domain (DBD)–FXR ligand-binding domain (LBD) chimeric receptor along with a Gal4-responsive luciferase reporter responded in the expected fashion when treated with the bile acid ligand, chenodeoxycholic acid (CDCA). CDCA activated the chimeric receptor with the expected EC₅₀ in the range of 40 μM; surprisingly however, we also noted that T0901317 activated FXR with an EC₅₀ of ∼5 μM. Although the FXR potency of T0901317 is

Discussion

LXR was initially identified as an orphan member of the nuclear receptor superfamily in 1995 [13]; however, its role as the physiological receptor for oxidized metabolites of cholesterol was rapidly elucidated [14], [15]. The natural oxysterol ligands were not extremely useful for characterization of LXR function due to their relatively low affinity and pleiotrophic actions. The discovery of the first high affinity synthetic LXR agonist, T0901317, provided a critical tool for characterization

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Citation Excerpt :
In addition to cross-talk, cross-reactivity between LXR and other NHRs is not uncommon. T0 is used as a benchmark LXR agonist; however, it has off-target activity at FXR382. The LXR agonist GSK2033 binds to PPARs366.
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T0901317 is a dual LXR/FXR agonist

Abstract

Introduction

Section snippets

Cell culture and transfections

Results

Discussion

Cell

J. Lipid Res.

Mol. Genet. Metab.

J. Biol. Chem.

J. Biol. Chem.

Mol. Cell

Mol. Cell

J. Biol. Chem.

J. Hepatol.