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A Gpr120-selective agonist improves insulin resistance and chronic inflammation in obese mice.

Oh da Y, Walenta E, Akiyama TE, Lagakos WS, Lackey D, Pessentheiner AR, Sasik R, Hah N, Chi TJ, Cox JM, Powels MA, Di Salvo J, Sinz C, Watkins SM, Armando AM, Chung H, Evans RM, Quehenberger O, McNelis J, Bogner-Strauss JG, Olefsky JM - Nat. Med. (2014)

Bottom Line: It is well known that the ω-3 fatty acids (ω-3-FAs; also known as n-3 fatty acids) can exert potent anti-inflammatory effects.We reported that Gpr120 is the functional receptor for these fatty acids and that ω-3-FAs produce robust anti-inflammatory, insulin-sensitizing effects, both in vivo and in vitro, in a Gpr120-dependent manner.However, the amount of fish oils that would have to be consumed to sustain chronic agonism of Gpr120 is too high to be practical, and, thus, a high-affinity small-molecule Gpr120 agonist would be of potential clinical benefit.

View Article: PubMed Central - PubMed

Affiliation: Division of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, La Jolla, California, USA.

ABSTRACT
It is well known that the ω-3 fatty acids (ω-3-FAs; also known as n-3 fatty acids) can exert potent anti-inflammatory effects. Commonly consumed as fish products, dietary supplements and pharmaceuticals, ω-3-FAs have a number of health benefits ascribed to them, including reduced plasma triglyceride levels, amelioration of atherosclerosis and increased insulin sensitivity. We reported that Gpr120 is the functional receptor for these fatty acids and that ω-3-FAs produce robust anti-inflammatory, insulin-sensitizing effects, both in vivo and in vitro, in a Gpr120-dependent manner. Indeed, genetic variants that predispose to obesity and diabetes have been described in the gene encoding GPR120 in humans (FFAR4). However, the amount of fish oils that would have to be consumed to sustain chronic agonism of Gpr120 is too high to be practical, and, thus, a high-affinity small-molecule Gpr120 agonist would be of potential clinical benefit. Accordingly, Gpr120 is a widely studied drug discovery target within the pharmaceutical industry. Gpr40 is another lipid-sensing G protein-coupled receptor, and it has been difficult to identify compounds with a high degree of selectivity for Gpr120 over Gpr40 (ref. 11). Here we report that a selective high-affinity, orally available, small-molecule Gpr120 agonist (cpdA) exerts potent anti-inflammatory effects on macrophages in vitro and in obese mice in vivo. Gpr120 agonist treatment of high-fat diet-fed obese mice causes improved glucose tolerance, decreased hyperinsulinemia, increased insulin sensitivity and decreased hepatic steatosis. This suggests that Gpr120 agonists could become new insulin-sensitizing drugs for the treatment of type 2 diabetes and other human insulin-resistant states in the future.

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CpdA is a selective agonist of Gpr120(a) Chemical structure of cpdA. Dose response data for cpdA are shown in (b) Ca2+ mobilization for Gpr120 vs. Gpr40 transfected cells. Dose response data for cpdA are shown in (c) IP3 production, and (d) β-arrestin-2 interaction assay with human and mouse Gpr120. Results are % activity over basal. (e) Gpr120-mediated SRE-luc activity after treatment with DHA and cpdA for 6 hr in HEK 293 cells. (f) NFkB-luc activity after pretreatment with DHA and cpdA for 1 hr subjected followed by LPS for 6 hr in primary macrophages from WT or Gpr120 KO mice. Results are fold activities over basal. Each data point represents mean±SEM of three independent experiments performed in triplicate. P<0.05 versus LPS treatment in WT macrophages. (g) DHA and cpdA inhibits LPS-induced inflammatory signaling in primary macrophages from WT, but not Gpr120 KO macrophages. The scanned bar graph (right panel) shows fold induction over basal conditions (p-Tak, p-Ikk, and p-Jnk) or LPS treatment (IkB degradation). Data are expressed as the mean±SEM. *, P<0.05 versus LPS treatment in WT mice to DHA+LPS or cpdA+LPS. n=6 per group. Data is a representative image from more than five independent experiments.
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Figure 1: CpdA is a selective agonist of Gpr120(a) Chemical structure of cpdA. Dose response data for cpdA are shown in (b) Ca2+ mobilization for Gpr120 vs. Gpr40 transfected cells. Dose response data for cpdA are shown in (c) IP3 production, and (d) β-arrestin-2 interaction assay with human and mouse Gpr120. Results are % activity over basal. (e) Gpr120-mediated SRE-luc activity after treatment with DHA and cpdA for 6 hr in HEK 293 cells. (f) NFkB-luc activity after pretreatment with DHA and cpdA for 1 hr subjected followed by LPS for 6 hr in primary macrophages from WT or Gpr120 KO mice. Results are fold activities over basal. Each data point represents mean±SEM of three independent experiments performed in triplicate. P<0.05 versus LPS treatment in WT macrophages. (g) DHA and cpdA inhibits LPS-induced inflammatory signaling in primary macrophages from WT, but not Gpr120 KO macrophages. The scanned bar graph (right panel) shows fold induction over basal conditions (p-Tak, p-Ikk, and p-Jnk) or LPS treatment (IkB degradation). Data are expressed as the mean±SEM. *, P<0.05 versus LPS treatment in WT mice to DHA+LPS or cpdA+LPS. n=6 per group. Data is a representative image from more than five independent experiments.

Mentions: Gpr120 and Gpr40 are 2 lipid sensing G protein-coupled receptors (GPCRs) 10,12, but despite limited homology between these two polyunsaturated fatty acid (PUFA) receptors, identification of ligands that are highly selective for Gpr120 over Gpr40 has been challenging 11,13-15. We have generated a small molecule Gpr120 agonist, compound A (cpdA) (Fig. 1a), and have examined its selectivity for Gpr120 compared to Gpr40 using a Ca2+ FLIPR assay (Fig. 1b). CpdA was fully selective for Gpr120 (logEC50 (M) = −7.62 ± 0.11) with negligible activity towards Gpr40 (Fig. 1b). Gpr120 couples to Gαq/11-initiated signal transduction pathways, and, as such, we assessed the activity of cpdA in an inositol-1, 4, 5-triphosphate (IP3) production assay, employing HEK 293 cells that stably express human or mouse Gpr120. The Gpr120 agonist produced concentration dependent increases in IP3 production from both human and mouse Gpr120 expressing cells (Fig. 1c). In addition to promoting signaling via Gαq/11, Gpr120 also directly couples to β-arrestin-2 8,14. Therefore, we examined the potency of cpdA in a β-arrestin-2 recruitment assay (Fig. 1d). CpdA led to a concentration-dependent response to recruit β-arrestin-2 in both human and mouse Gpr120 expressing cells, with EC50s of ~0.35 μM (Fig. 1d). Since Gpr120 is a Gαq/11-coupled receptor, it stimulates both PKC and MAP kinase, and both of these biologic effects can be detected in an SRE-driven reporter system 8. HEK293 cells were transiently transfected with constructs for mouse Gpr120 along with a serum response element-luciferase promoter/reporter (SRE-luc). The Gpr120 SRE-luc reporter cells were treated with docosahexaenoic acid (DHA) and cpdA. Gpr120 stimulation by cpdA was ~50 fold more potent than DHA (Fig. 1e). DHA and cpdA were used at 100 μM and 10 μM in all subsequent studies to achieve maximal effects.


A Gpr120-selective agonist improves insulin resistance and chronic inflammation in obese mice.

Oh da Y, Walenta E, Akiyama TE, Lagakos WS, Lackey D, Pessentheiner AR, Sasik R, Hah N, Chi TJ, Cox JM, Powels MA, Di Salvo J, Sinz C, Watkins SM, Armando AM, Chung H, Evans RM, Quehenberger O, McNelis J, Bogner-Strauss JG, Olefsky JM - Nat. Med. (2014)

CpdA is a selective agonist of Gpr120(a) Chemical structure of cpdA. Dose response data for cpdA are shown in (b) Ca2+ mobilization for Gpr120 vs. Gpr40 transfected cells. Dose response data for cpdA are shown in (c) IP3 production, and (d) β-arrestin-2 interaction assay with human and mouse Gpr120. Results are % activity over basal. (e) Gpr120-mediated SRE-luc activity after treatment with DHA and cpdA for 6 hr in HEK 293 cells. (f) NFkB-luc activity after pretreatment with DHA and cpdA for 1 hr subjected followed by LPS for 6 hr in primary macrophages from WT or Gpr120 KO mice. Results are fold activities over basal. Each data point represents mean±SEM of three independent experiments performed in triplicate. P<0.05 versus LPS treatment in WT macrophages. (g) DHA and cpdA inhibits LPS-induced inflammatory signaling in primary macrophages from WT, but not Gpr120 KO macrophages. The scanned bar graph (right panel) shows fold induction over basal conditions (p-Tak, p-Ikk, and p-Jnk) or LPS treatment (IkB degradation). Data are expressed as the mean±SEM. *, P<0.05 versus LPS treatment in WT mice to DHA+LPS or cpdA+LPS. n=6 per group. Data is a representative image from more than five independent experiments.
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Figure 1: CpdA is a selective agonist of Gpr120(a) Chemical structure of cpdA. Dose response data for cpdA are shown in (b) Ca2+ mobilization for Gpr120 vs. Gpr40 transfected cells. Dose response data for cpdA are shown in (c) IP3 production, and (d) β-arrestin-2 interaction assay with human and mouse Gpr120. Results are % activity over basal. (e) Gpr120-mediated SRE-luc activity after treatment with DHA and cpdA for 6 hr in HEK 293 cells. (f) NFkB-luc activity after pretreatment with DHA and cpdA for 1 hr subjected followed by LPS for 6 hr in primary macrophages from WT or Gpr120 KO mice. Results are fold activities over basal. Each data point represents mean±SEM of three independent experiments performed in triplicate. P<0.05 versus LPS treatment in WT macrophages. (g) DHA and cpdA inhibits LPS-induced inflammatory signaling in primary macrophages from WT, but not Gpr120 KO macrophages. The scanned bar graph (right panel) shows fold induction over basal conditions (p-Tak, p-Ikk, and p-Jnk) or LPS treatment (IkB degradation). Data are expressed as the mean±SEM. *, P<0.05 versus LPS treatment in WT mice to DHA+LPS or cpdA+LPS. n=6 per group. Data is a representative image from more than five independent experiments.
Mentions: Gpr120 and Gpr40 are 2 lipid sensing G protein-coupled receptors (GPCRs) 10,12, but despite limited homology between these two polyunsaturated fatty acid (PUFA) receptors, identification of ligands that are highly selective for Gpr120 over Gpr40 has been challenging 11,13-15. We have generated a small molecule Gpr120 agonist, compound A (cpdA) (Fig. 1a), and have examined its selectivity for Gpr120 compared to Gpr40 using a Ca2+ FLIPR assay (Fig. 1b). CpdA was fully selective for Gpr120 (logEC50 (M) = −7.62 ± 0.11) with negligible activity towards Gpr40 (Fig. 1b). Gpr120 couples to Gαq/11-initiated signal transduction pathways, and, as such, we assessed the activity of cpdA in an inositol-1, 4, 5-triphosphate (IP3) production assay, employing HEK 293 cells that stably express human or mouse Gpr120. The Gpr120 agonist produced concentration dependent increases in IP3 production from both human and mouse Gpr120 expressing cells (Fig. 1c). In addition to promoting signaling via Gαq/11, Gpr120 also directly couples to β-arrestin-2 8,14. Therefore, we examined the potency of cpdA in a β-arrestin-2 recruitment assay (Fig. 1d). CpdA led to a concentration-dependent response to recruit β-arrestin-2 in both human and mouse Gpr120 expressing cells, with EC50s of ~0.35 μM (Fig. 1d). Since Gpr120 is a Gαq/11-coupled receptor, it stimulates both PKC and MAP kinase, and both of these biologic effects can be detected in an SRE-driven reporter system 8. HEK293 cells were transiently transfected with constructs for mouse Gpr120 along with a serum response element-luciferase promoter/reporter (SRE-luc). The Gpr120 SRE-luc reporter cells were treated with docosahexaenoic acid (DHA) and cpdA. Gpr120 stimulation by cpdA was ~50 fold more potent than DHA (Fig. 1e). DHA and cpdA were used at 100 μM and 10 μM in all subsequent studies to achieve maximal effects.

Bottom Line: It is well known that the ω-3 fatty acids (ω-3-FAs; also known as n-3 fatty acids) can exert potent anti-inflammatory effects.We reported that Gpr120 is the functional receptor for these fatty acids and that ω-3-FAs produce robust anti-inflammatory, insulin-sensitizing effects, both in vivo and in vitro, in a Gpr120-dependent manner.However, the amount of fish oils that would have to be consumed to sustain chronic agonism of Gpr120 is too high to be practical, and, thus, a high-affinity small-molecule Gpr120 agonist would be of potential clinical benefit.

View Article: PubMed Central - PubMed

Affiliation: Division of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, La Jolla, California, USA.

ABSTRACT
It is well known that the ω-3 fatty acids (ω-3-FAs; also known as n-3 fatty acids) can exert potent anti-inflammatory effects. Commonly consumed as fish products, dietary supplements and pharmaceuticals, ω-3-FAs have a number of health benefits ascribed to them, including reduced plasma triglyceride levels, amelioration of atherosclerosis and increased insulin sensitivity. We reported that Gpr120 is the functional receptor for these fatty acids and that ω-3-FAs produce robust anti-inflammatory, insulin-sensitizing effects, both in vivo and in vitro, in a Gpr120-dependent manner. Indeed, genetic variants that predispose to obesity and diabetes have been described in the gene encoding GPR120 in humans (FFAR4). However, the amount of fish oils that would have to be consumed to sustain chronic agonism of Gpr120 is too high to be practical, and, thus, a high-affinity small-molecule Gpr120 agonist would be of potential clinical benefit. Accordingly, Gpr120 is a widely studied drug discovery target within the pharmaceutical industry. Gpr40 is another lipid-sensing G protein-coupled receptor, and it has been difficult to identify compounds with a high degree of selectivity for Gpr120 over Gpr40 (ref. 11). Here we report that a selective high-affinity, orally available, small-molecule Gpr120 agonist (cpdA) exerts potent anti-inflammatory effects on macrophages in vitro and in obese mice in vivo. Gpr120 agonist treatment of high-fat diet-fed obese mice causes improved glucose tolerance, decreased hyperinsulinemia, increased insulin sensitivity and decreased hepatic steatosis. This suggests that Gpr120 agonists could become new insulin-sensitizing drugs for the treatment of type 2 diabetes and other human insulin-resistant states in the future.

Show MeSH
Related in: MedlinePlus