The Inverse Agonist DG172 Triggers a PPARβ/δ-Independent Myeloid Lineage Shift and Promotes GM-CSF/IL-4-Induced Dendritic Cell Differentiation

• Published in: Molecular pharmacology 2015-02, Vol.87 (2), p.162-173
• Main Authors: Lieber, Sonja, Scheer, Frithjof, Finkernagel, Florian, Meissner, Wolfgang, Giehl, Gavin, Brendel, Cornelia, Diederich, Wibke E., Müller-Brüsselbach, Sabine, Müller, Rolf
• Format: Article
• Language: English
• Subjects: (Z)-2-(2-bromophenyl)-3-{[4-(1-methylpiperazine)amino]phenyl}acrylonitrile; (Z)-2-(4-chlorophenyl)-3-[4-(4-methylpiperazine-1-yl)phenyl]acrylonitrile; Acrylonitrile - analogs & derivatives; Acrylonitrile - pharmacology; Animals; BMC; bone marrow cell; Bone Marrow Cells - drug effects; Bone Marrow Cells - metabolism; Cell Differentiation - drug effects; Cell Differentiation - physiology; Cell Lineage; Cells, Cultured; dendritic cell; Dendritic Cells - drug effects; Dendritic Cells - metabolism; DG172; DG228; Drug Inverse Agonism; Drug Synergism; FACS; FITC; fluorescein isothiocyanate; fluorescence-activated cell sorting; GM-CSF; Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology; granulocyte-macrophage-colony-stimulating factor; GW501516; Interleukin-4 - pharmacology; lipopolysaccharide; LPS; M-CSF; macrophage-colony-stimulating factor; mean fluorescence intensity; methyl 3-{N-[4-(hexylamino)-2-methoxyphenyl]sulfamoyl}thiophene-2-carboxylate; MFI; Mice; Mice, Inbred C57BL; Mice, Knockout; peroxisome proliferator-activated receptor; Piperazines - pharmacology; PPAR; PPAR gamma - agonists; PPAR gamma - metabolism; PPAR-beta - agonists; PPAR-beta - metabolism; real-time quantitative polymerase chain reaction; RT-qPCR; ST247; time-resolved fluorescence resonance energy transfer; TR-FRET; wild-type; {2-methyl-4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-5-thiazolyl}methyl)thio]phenoxy} acetic acid
• ISSN: 0026-895X; 1521-0111
• DOI: 10.1124/mol.114.094672

The stilbene derivative (Z)-2-(2-bromophenyl)-3-{[4-(1-methylpiperazine)amino]phenyl}acrylonitrile (DG172) was developed as a highly selective inhibitory peroxisome proliferator-activated receptor (PPAR)β/δ ligand. Here, we describe a novel PPARβ/δ-independent, yet highly specific, effect of DG172 on the differentiation of bone marrow cells (BMCs). DG172 strongly augmented granulocyte-macrophage-colony-stimulating factor (GM-CSF)-induced differentiation of primary BMCs from Ppard null mice into two specific populations, characterized as mature (CD11chiMHCIIhi) and immature (CD11chiMHCIIlo) dendritic cells (DCs). IL-4 synergized with DG172 to shift the differentiation from MHCIIlo cells to mature DCs in vitro. The promotion of DC differentiation occurred at the expense of differentiation to granulocytic Gr1+Ly6B+ cells. In agreement with these findings, transcriptome analyses showed a strong DG172-mediated repression of genes encoding neutrophilic markers in both differentiating wild-type and Ppard null cells, while macrophage/DC marker genes were up-regulated. DG172 also inhibited the expression of transcription factors driving granulocytic differentiation (Cebpe, Gfi1, and Klf5), and increased the levels of transcription factors promoting macrophage/DC differentiation (Irf4, Irf8, Spib, and Spic). DG172 exerted these effects only at an early stage of BMC differentiation induced by GM-CSF, did not affect macrophage-colony-stimulating factor–triggered differentiation to macrophages and had no detectable PPARβ/δ-independent effect on other cell types tested. Structure-function analyses demonstrated that the 4-methylpiperazine moiety in DG172 is required for its effect on DC differentiation, but is dispensable for PPARβ/δ binding. Based on these data we developed a new compound, (Z)-2-(4-chlorophenyl)-3-[4-(4-methylpiperazine-1-yl)phenyl]acrylonitrile (DG228), which enhances DC differentiation in the absence of significant PPARβ/δ binding.