gdf15  (R&D Systems)

Journal: Italian Journal of Pediatrics

Article Title: Increased serum carboxylesterase-1 levels are associated with metabolic dysfunction associated steatotic liver disease and metabolic syndrome in children with obesity

doi: 10.1186/s13052-024-01733-7

Figure Lengend Snippet: Characteristics of obesity subjects according to MASLD and MetS

Article Snippet: Serum levels of adiponectin, leptin, GDF15, IGF-1, CES1 were measured using enzyme-linked immunosorbent assay ELESA kits (Boster Biological, Pleasanton, CA) following the manufacturer’s protocol.

Techniques:

Journal: Italian Journal of Pediatrics

Article Title: Increased serum carboxylesterase-1 levels are associated with metabolic dysfunction associated steatotic liver disease and metabolic syndrome in children with obesity

doi: 10.1186/s13052-024-01733-7

Figure Lengend Snippet: Correlation analysis between CES1 and obesity-related secreted proteins. ( A ) Adiponectin, ( B ) Leptin, ( C ) GDF15, ( D ) IGF-1. Spearman correlation coefficient was used. Statistical assessment was 2-sided and considered statistically significant at * P < 0.05, ** P < 0.01

Article Snippet: Serum levels of adiponectin, leptin, GDF15, IGF-1, CES1 were measured using enzyme-linked immunosorbent assay ELESA kits (Boster Biological, Pleasanton, CA) following the manufacturer’s protocol.

Techniques:

Journal: Nature

Article Title: PTER is a N -acetyltaurine hydrolase that regulates feeding and obesity

doi: 10.1038/s41586-024-07801-6

Figure Lengend Snippet: a , b , Change in body weight ( a ) and food intake ( b ) of 26–28-week-old male DIO C57BL/6J mice following 7 days of treatment with the indicated dose of N -acetyltaurine (NAT; i.p.). For saline versus N -acetyltaurine (15 mg per kg per day), P = 5.95 × 10 –4 ; for saline versus N -acetyltaurine (50 mg per kg per day), P = 6.3 × 10 –4 . N = 5 per group for vehicle, 1 and 5 mg per kg per day; N = 6 per group for 15 and 50 mg per kg per day. c , d , Change in body weight ( c ) and food intake ( d ) of 19–21-week-old male DIO C57BL/6J mice following treatment with the indicated metabolite at a dose of 15 mg per kg per day (i.p.). N = 5 per group. e – g , Western blots with anti-PTER (top) and anti-tubulin (bottom) antibodies ( e ), N -acetyltaurine hydrolysis activity ( f ) and tissue N -acetyltaurine levels ( g ) from cortex (Cort.), hypothalamus (Hyp.) and brainstem (BS) of WT mice and Pter KO mice. For WT versus Pter KO brainstem, P = 6.65 × 10 –4 . N = 6 per group for f and g . h , Change in 24-h food intake of 6-month-old male DIO mice treated with a single dose of GDF15 (0.1 mg kg –1 , i.p.) in the presence of anti-GFRAL antibody (10 mg kg –1 , i.p.) or IgG control antibody (10 mg kg –1 , i.p.). N = 5 per group. i , j , Change in body weight ( i ) and cumulative food intake ( j ) of 16-week-old male DIO mice following saline or N -acetyltaurine (15 mg per kg per day, i.p.) treatment and with IgG or anti-GFRAL antibody co-treatment (10 mg kg –1 , i.p., once every 3 days). N = 10 per group. Data are shown as the mean ± s.e.m. For e , the loading control was performed on the same blot. In a – d and f – h , P values were calculated from two-tailed unpaired t -tests and were not corrected for multiple comparisons. In i and j , P values were calculated from two-way ANOVA with post hoc Sidak’s multiple comparisons test. All experiments were performed once.

Article Snippet: Recombinant GDF15 (957-GD) was purchased from R&D Systems.

Techniques: Saline, Western Blot, Activity Assay, Control, Two Tailed Test

Journal: Nature

Article Title: PTER is a N -acetyltaurine hydrolase that regulates feeding and obesity

doi: 10.1038/s41586-024-07801-6

Figure Lengend Snippet: Related to Fig. . a - d , Body weight ( a , b ) and food intake ( c , d ) of 6 to 7-month-old DIO male C57BL/6J mice after a 7-day treatment of saline or N-acetyltaurine (15 mg/kg/day, IP) or GLP-1 (2 mg/kg/day, IP) with or without Exendin-3 (0.1 mg/kg/day, IP). N = 7/group. NAT, N-acetyltaurine. e , f , Body weight ( e ) and food intake ( f ) of 5-month-old DIO male C57BL/6J mice or 3 to 4-month-old MC4R-KO mice on high fat diet after a 7-day treatment of saline or N-acetyltaurine (15 mg/kg/day, IP). N = 6/group. NAT, N-acetyltaurine. g , Plasma GDF15 (left), GLP-1 (middle) and leptin (right) levels of 19 to 21-week-old male DIO C57BL/6J mice following treatment with N-acetyltaurine 15 mg/kg/day (IP) or saline. N = 5 per group. NAT, N-acetyltaurine. Data are shown as mean ± SEM. P-values were calculated from two-tailed unpaired t-tests.

Article Snippet: Recombinant GDF15 (957-GD) was purchased from R&D Systems.

Techniques: Saline, Two Tailed Test

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 1: Effect of oral intake of Plaquenil on GDF15 and ghrelin plasma levels, and hunger scores in healthy volunteers. A) Time-dependent changes in plasma GDF15 levels after oral ingestion of Plaquenil or placebo in healthy volunteers (n ¼ 10; *P < 0.05: versus baseline [10 min], ##P < 0.01: versus the placebo condition; ANCOVA mixed model). Dashed line: trendline in the fasted state; dotted line: extrapolation from trendline in the fasted state. B and C) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and hungers scores measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values). D and E) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and ghrelin plasma levels measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values).All data were presented as mean SEM.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Clinical Proteomics, Transformation Assay

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 2: Distribution and characterization of GDF15 expressing epithelial cells in the proximal gut of normal-weight individuals and patients with obesity. A) Relative mRNA expression (efficiencyDDCt method) of GDF15 in resection specimens from the fundus (nNW ¼ 7, nOB ¼ 10), corpus (nNW ¼ 5, nOB ¼ 8), antrum (nNW ¼ 5, nOB ¼ 8), and jejunum (nNW ¼ 5, nOB ¼ 7) of normal-weight individuals and patients with obesity. (Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 in jejunal sections (10 mm) of a normal-weight individual and a patient with obesity. GDF15þ cells were stained with Cy3 (green) and nuclei were stained with DAPI (blue). Scale bars: 25 mm. C) Average number and intensity of GDF15þ cells in jejunal tissue sections from both normal-weight individuals and patients with obesity (nNW ¼ 5, nOB ¼ 5; two-tailed unpaired Student’s t-test). DeH) Representative double-immunofluorescence staining for GDF15 (green [Cy3 or Alexa594]) with markers for (D) goblet (MUC2: red [Cy5]), (E) Paneth (a-defensin 6: red [Cy5]) or (FeG) enteroendocrine (CHGA: red [Alexa488]) and ghrelin (red [Alexa488]) cells in jejunal sections (10 mm) from normal-weight individuals. Arrows indicate co-localization. Normal rabbit serum was used as negative control. Nuclei were labeled with DAPI (blue). Scale bars: 25 mm. NW: normal-weight, OB: obese. Data of figure A, C represent mean SEM and single values are plotted. *P < 0.05: versus jejunum in normal-weight individuals, $$$P < 0.001: versus jejunum in patients with obesity, #P < 0.05, ###P < 0.001: versus normal-weight individuals.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Expressing, Staining, Two Tailed Test, Negative Control, Labeling

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 3: Effects of bitter generalists and intermediates on GDF15 or GLP-1 expression in jejunal crypts from patients with obesity. A) Effect of 4-hour stimulation with 100 mM HCQS on relative GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). B) Effect of 4-hour stimulation with 100 mM HCQS on relative GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). C) Effect of bitter generalists on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 6e9; Proc Mixed Model with Sidák correction for multiple comparisons). D) Effect of bitter intermediates on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e6; Proc Mixed Model with Sidák correction for multiple comparisons). E) Relative GDF15 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin. Azithromycin was removed after 4 h for the 24-hour timepoint (n ¼ 3e6; one-tailed paired Student’s t-test). F) Relative GLP-1 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin for 6 h (n ¼ 3; two-tailed paired Student’s t-test). All data are present as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus the vehicle group.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Expressing, Two Tailed Test, One-tailed Test

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 4: Effects of bitter specialists on GDF15 mRNA or protein expression in primary jejunal crypts from patients with obesity. A) Effect of specialists on relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e7; Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 (green [Cy3]) in primary crypts treated with 1 mM gallic acid or vehicle after 24 h with 4 h stimulation. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. C) Average intensity of GDF15þ cells in primary crypts treated with 1 mM gallic acid or vehicle. Gallic acid was removed after 4 h of stimulation and staining was performed 20 h later (n ¼ 4; two-tailed paired Student’s t-test). D) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen for 6 h (n ¼ 3; one-tailed paired Student’s t-test). E) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen. Acetaminophen was removed after 4 h for the 24-hour timepoint (n ¼ 3; one-tailed paired Student’s t-test). F) Venn diagram of bitter compounds that affected GDF15 and/or GLP-1 mRNA expression after stimulation or primary crypts for 4 h. Asterisk (*) indicates an inhibitory effect on mRNA expression; the use of hashtags (#) signifies potential variations in the effects on GDF15 or GLP-1 mRNA, depending on the genotype of the patients. Data of figure A, C, D, E are present as mean SEM and single values are plotted. **P < 0.01, ***P < 0.001: versus the vehicle group.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Expressing, Staining, Labeling, Two Tailed Test, One-tailed Test

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 5: Role of TAS2Rs and the motilin receptor in the effect of bitter agonists on GDF15 or GLP-1 expression in primary crypts from patients with obesity. A) The TAS2R antagonist, GIV3727 (110 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). B) GIV3727 (110 mM) did not block the effect of gallic acid (1 mM) on GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). C) GIV3727 (110 mM) did not block the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). D) The motilin receptor antagonist MA-2029 blocked the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). E) C12-O-AHL (0.15 mM) increased relative GDF15 mRNA expression in primary crypts from obese patients with TAS2R4(GG/CG) (n ¼ 9) but not TAS2R4(CC) (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). F) C12-O-AHL (0.15 mM) decreased GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R4(GG/CG) (n ¼ 9) but not the TAS2R4(CC) genotype (n ¼ 4). (Proc Mixed Model with Sidák correction for multiple comparisons). G) Aloin at 30 mM (white dots) and 100 mM (black dots) decreased relative GDF15 mRNA expression in primary crypts from obese patients with the TAS2R43(þ)GG/CG (n ¼ 9) but not the TAS2R43(þ)CC (n ¼ 6) or TAS2R43() (n ¼ 6) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). H) Aloin at 30 mM (white dots) and 100 mM (black dots) did not affect the relative GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R43(þ) GG/GC (n ¼ 9), TAS2R43(þ) CC (n ¼ 6), or TAS2R43() (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). IeK) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in jejunal sections (10 mm thickness) from normal-weight populations. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. MeO) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in primary jejunal crypt from patients with obesity. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. (L and P) Normal rabbit serum as a negative control. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. Data of figure AeH are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01, ###P < 0.001: bitter treatment antagonist, $P < 0.05: versus TAS2R4(CC) population, &P < 0.05: versus TAS2R43() population.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Expressing, Blocking Assay, Labeling, Negative Control

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 6: Role of the unfolded protein response pathway in the effect of bitter agonists on GDF15 and GLP1 mRNA expression in primary crypts from patients with obesity. A) Positive correlation between Log2 fold change of GDF15 and DDIT3 mRNA expression induced by bitter agonists (bitter: DB, AZI, GA, PHE, EM, ACE, BERB; n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). B) Positive correlation between Log2 fold change of GDF15 and ATF4 mRNA expression induced by bitter agonists from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). C) Positive correlation between Log2 fold change of DDIT3 and ATF4 mRNA expression induced by bitter agonist from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). D) ISRIB (5 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). E) ISRIB (5 mM) did not affect the effects of gallic acid (1 mM) on GLP-1 mRNA expression (n ¼ 2; Proc Mixed Model with Sidák correction for multiple comparisons). Data are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01: bitter treatment antagonist.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Expressing