Research ArticleMetabolic Disease

Preclinical efficacy of the GPER-selective agonist G-1 in mouse models of obesity and diabetes

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Science Translational Medicine  29 Jan 2020:
Vol. 12, Issue 528, eaau5956
DOI: 10.1126/scitranslmed.aau5956
  • Fig. 1 Selective activation of GPER in OVX mice attenuates obesity.

    Body weight in OVX mice after treatment with GPER agonist G-1 compared to vehicle (Veh) controls and ovary-intact mice (A) over time and (B) at termination of the study. Images of representative mice are shown in (B). The weight of the perigonadal (C) and perirenal (D) fat pads, circulating cholesterol (E), and uterine wet weight (F) were determined at termination of the study. (A) n = 5 (intact) to 14 (OVX + Veh and OVX + G-1), (B) n = 13, (C, D, and F) n = 7, and (E) n = 6. (A) Two-way ANOVA; *P < 0.05, **P < 0.01, and ***P < 0.001 for vehicle treatment versus ovary intact controls, respectively; #P < 0.05 for G-1 treatment versus vehicle treatment. (B to F) One-way ANOVA with Bonferroni post hoc test.

  • Fig. 2 Treatment with GPER-selective agonist G-1 reduces fat content in OVX mice.

    (A) Representative DEXA scans, (B) overall body fat content, (C) body fat percentage, (D) lean mass, (E) bone mineral density, and (F) bone mineral content in vehicle- and G-1–treated OVX mice compared to ovary-intact animals. Representative anatomical MRI images of the (G) coronal and (I) axial view in different mouse cohorts with quantification of (H) total (from coronal view) and (J) subcutaneous (from axial view) fat content. (B to F) n = 6 and (H and J) n = 4. All tests are one-way ANOVA with Bonferroni post hoc test.

  • Fig. 3 GPER activation in OVX mice increases energy expenditure.

    (A) Energy expenditure over time (VO2; milliliter/hour), (B) RER over time (VCO2/VO2), (C) oxygen consumption (VO2; total, light and dark phases), (D) food intake, and (E) locomotor activity over time and (F) total, light and dark phases, in vehicle- and G-1–treated OVX mice compared to the ovary-intact control animals. (A to C) n = 6 (intact) to 8 (OVX + Veh and OVX + G-1) and (D to F) n = 4 to 7. All tests are one-way ANOVA with Bonferroni post hoc test.

  • Fig. 4 Adipose tissue remodeling in OVX mice treated with GPER-selective agonist G-1.

    (A) Perigonadal fat pads (whole mount and H&E-stained sections; scale bar, 100 μm) and (B) mean adipocyte area as quantified after H&E staining in (A) (n = 5). (C) Gene expression analyses in perigonadal WAT for genes involved in angiogenesis (Hif1a and Vegfa) and mitochondrial biogenesis (Ppargc1a) and fatty acid oxidation (Acox1) (n = 6). (D) Gene expression analyses in BAT for genes involved in thermogenesis (Ucp1) and sympathetic innervation (Th) (n = 6). Statistical significance was determined by one-way ANOVA with Bonferroni post hoc test.

  • Fig. 5 G-1 treatment increases mitochondrial gene expression and cellular respiration.

    Expression of the mitochondrial genes Ppargc1a, Acaca, Nrf1, Tfam, and Acox1 in (A) BAT and (B) skeletal muscle after vehicle or G-1 treatment of OVX mice (n = 5 to 6). (C) Oxygen consumption rate (OCR) of brown preadipocytes under basal conditions (0 to 16 min) after GPER stimulation for 24 hours with 100 nM G-1.*P < 0.001 for G-1–treated cells versus vehicle control. (D) Basal OCR, maximal OCR, spare respiratory capacity, and OCR for ATP production in G-1–treated cells versus control cells. The results shown in (C) and (D) are representative of three independent experiments with four replicates for each condition per experiment as indicated. Statistical significance was determined by (A and B) one-way ANOVA with Bonferroni post hoc test, (C) two-way ANOVA, and (D) Mann-Whitney U test.

  • Fig. 6 GPER agonism attenuates inflammation resulting from OVX.

    (A) Systemic concentrations of inflammatory cytokines (n = 8). Gene expression of inflammatory markers in (B) perigonadal WAT, (C) liver, and (D) skeletal muscle (n = 5 to 6). Statistical significance was determined by one-way ANOVA with Bonferroni post hoc test.

  • Fig. 7 GPER-selective agonist G-1 improves glucose homeostasis in OVX mice and modulates systemic concentrations of metabolic hormones.

    (A and B) Tolerance to glucose (glucose tolerance test, GTT); fasting plasma (C), glucose, and (D) insulin; and (E) HOMA-IR in vehicle- and G-1–treated OVX mice compared to ovary-intact mice. For GTT, area under the curve for each individual mouse in (A) was plotted in (B). (F) Concentrations of plasma leptin, insulin, pancreatic polypeptide (PP), C-peptide 2, and glucagon in the fed state. (A to E) n = 6 and (F) n = 6 to 8. Statistical significance was determined by one-way ANOVA with Bonferroni post hoc test.

  • Fig. 8 Activation of GPER by G-1 exerts antiobesity and antidiabetic effects in male DIO mice.

    (A) Body weights (n = 14), (B) plasma cholesterol concentrations (n = 6), (C) energy expenditure (n = 4), fasting (D) glucose (n = 6) and (E) insulin (n = 6) concentrations, (F) HOMA-IR (n = 6), and (G and H) glucose tolerance (n = 8) in DIO male mice treated with vehicle or G-1 compared in male mice fed normal chow (NC). For (D) and (H), area under the curve was determined for each individual mouse in (C) and (G), respectively. Statistical significance was determined by one-way ANOVA with Bonferroni post hoc test.

Supplementary Materials

  • stm.sciencemag.org/cgi/content/full/12/528/eaau5956/DC1

    Fig. S1. Baseline glucose homeostasis in female mice upon ovariectomy.

    Fig. S2. Plasma concentrations of ghrelin, PYY, GIP, amylin, and GLP-1 in OVX female mice after G-1 treatment.

    Fig. S3. Baseline glucose homeostasis in diet-induced obese male mice.

    Fig. S4. Body weights and glucose tolerance in OVX female mice on HFD after treatment with G-1.

    Fig. S5. Effects of different doses of G-1 on body weight in male mice with DIO.

    Fig. S6. Effects of G-1 dosing regimen on glucose tolerance in male mice with DIO.

    Table S1. Primers used for qPCR.

    Data file S1. Raw data from figures.

  • The PDF file includes:

    • Fig. S1. Baseline glucose homeostasis in female mice upon ovariectomy.
    • Fig. S2. Plasma concentrations of ghrelin, PYY, GIP, amylin, and GLP-1 in OVX female mice after G-1 treatment.
    • Fig. S3. Baseline glucose homeostasis in diet-induced obese male mice.
    • Fig. S4. Body weights and glucose tolerance in OVX female mice on HFD after treatment with G-1.
    • Fig. S5. Effects of different doses of G-1 on body weight in male mice with DIO.
    • Fig. S6. Effects of G-1 dosing regimen on glucose tolerance in male mice with DIO.
    • Table S1. Primers used for qPCR.
    • Legend for data file S1

    [Download PDF]

    Other Supplementary Material for this manuscript includes the following:

    • Data file S1 (Microsoft Excel format). Raw data from figures.

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