作者: Anderson, Aimee L ; Devereaux, Michael W ; Sokol, Ronald J ; Lovell, Mark ; El Kasmi, Karim C ; Orlicky, David J ; Shearn, Colin T ; Reisz, Julie A ; Ghosh, Swati ; D'Alessandro, Angelo ; Gehrke, Sarah

Background and Aims:

Chronically administered parenteral nutrition (PN) in patients with intestinal failure carries the risk for developing PN‐associated cholestasis (PNAC). We have demonstrated that farnesoid X receptor (FXR) and liver X receptor (LXR), proinflammatory interleukin‐1 beta (IL‐1β), and infused phytosterols are important in murine PNAC pathogenesis. In this study we examined the role of nuclear receptor liver receptor homolog 1 (LRH‐1) and phytosterols in PNAC.

Approach and Results:

In a C57BL/6 PNAC mouse model (dextran sulfate sodium [DSS] pretreatment followed by 14 days of PN; DSS‐PN), hepatic nuclear receptor subfamily 5, group A, member 2/LRH‐1 mRNA, LRH‐1 protein expression, and binding of LRH‐1 at the Abcg5/8 and Cyp7a1 promoter was reduced. Interleukin‐1 receptor–deficient mice (Il‐1r−/−/DSS‐PN) were protected from PNAC and had significantly increased hepatic mRNA and protein expression of LRH‐1. NF‐κB activation and binding to the LRH‐1 promoter were increased in DSS‐PN PNAC mice and normalized in Il‐1r−/−/DSS‐PN mice. Knockdown of NF‐κB in IL‐1β–exposed HepG2 cells increased expression of LRH‐1 and ABCG5. Treatment of HepG2 cells and primary mouse hepatocytes with an LRH‐1 inverse agonist, ML179, significantly reduced mRNA expression of FXR targets ATP binding cassette subfamily C member 2/multidrug resistance associated protein 2 (ABCC2/MRP2), nuclear receptor subfamily 0, groupB, member 2/small heterodimer partner (NR0B2/SHP), and ATP binding cassette subfamily B member 11/bile salt export pump (ABCB11/BSEP). Co‐incubation with phytosterols further reduced expression of these genes. Similar results were obtained by suppressing the LRH‐1 targets ABCG5/8 by treatment with small interfering RNA, IL‐1β, or LXR antagonist GSK2033. Liquid chromatography–mass spectrometry and chromatin immunoprecipitation experiments in HepG2 cells showed that ATP binding cassette subfamily G member 5/8 (ABCG5/8) suppression by GSK2033 increased the accumulation of phytosterols and reduced binding of FXR to the SHP promoter. Finally, treatment with LRH‐1 agonist, dilauroyl phosphatidylcholine (DLPC) protected DSS‐PN mice from PNAC.

Conclusions:

This study suggests that NF‐κB regulation of LRH‐1 and downstream genes may affect phytosterol‐mediated antagonism of FXR signaling in the pathogenesis of PNAC. LRH‐1 could be a potential therapeutic target for PNAC.

2020-08-01·Bioorganic & medicinal chemistry letters4区 · 医学

Development of a new class of liver receptor homolog-1 (LRH-1) agonists by photoredox conjugate addition

4区 · 医学

Article

作者: Jui, Nathan T ; Cornelison, Jeffery L ; Mays, Suzanne G ; D'Agostino, Emma H ; Houtman, René ; Ortlund, Eric A ; Cato, Michael L ; Johnson, Alyssa M ; Melchers, Diana ; Patel, Anamika B

LRH-1 is a nuclear receptor that regulates lipid metabolism and homeostasis, making it an attractive target for the treatment of diabetes and non-alcoholic fatty liver disease. Building on recent structural information about ligand binding from our labs, we have designed a series of new LRH-1 agonists that further engage LRH-1 through added polar interactions. While the current synthetic approach to this scaffold has, in large part, allowed for decoration of the agonist core, significant variation of the bridgehead substituent is mechanistically precluded. We have developed a new synthetic approach to overcome this limitation, identified that bridgehead substitution is necessary for LRH-1 activation, and described an alternative class of bridgehead substituents for effective LRH-1 agonist development. We determined the crystal structure of LRH-1 bound to a bridgehead-modified compound, revealing a promising opportunity to target novel regions of the ligand binding pocket to alter LRH-1 target gene expression.

2020-03-12·ACS medicinal chemistry letters3区 · 医学

Development of a Versatile and Sensitive Direct Ligand Binding Assay for Human NR5A Nuclear Receptors

3区 · 医学

Article

作者: Ortlund, Eric A ; Cornelison, Jeffery L ; Jui, Nathan T ; Patel, Anamika ; Mays, Suzanne G ; D'Agostino, Emma H ; Flynn, Autumn R

As regulators of steroidogenesis, development, and metabolism, the nuclear receptor 5A (NR5A) subfamily members steroidogenic factor 1 (SF-1) and liver receptor homologue 1 (LRH-1) are important pharmacological targets for cancers and metabolic diseases. Evaluation of small molecule modulators and candidate endogenous ligands for these orphan receptors has been hindered by the lack of accessible, robust direct-binding assays. Here, we leverage the potency of our new NR5A agonist (6N) to create a high-affinity probe for fluorescence polarization competition assays by conjugating 6N to fluorescein (FAM). The 6N-FAM probe tightly binds the NR5A receptors and detects direct binding of synthetic and phospholipid ligands. For 25 LRH-1 agonists, affinity predicts potency in cellular activation assays, demonstrating the potential for this assay in drug discovery. Moreover, phospholipids dilauroylphosphatidylcholine and phosphatidylinositol(4,5)phosphate bind with high affinity, demonstrating this assay is robust for evaluation of candidate endogenous ligands for human NR5A receptors.

100 项与 LRH-1 Inverse agonists (TES Pharma) 相关的药物交易

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