gdf15 levels  (Roche)

Journal: Molecular Metabolism

Article Title: Muscle weakness and mitochondrial stress occur before severe metastasis in a novel mouse model of ovarian cancer cachexia

doi: 10.1016/j.molmet.2024.101976

Figure Lengend Snippet: The effects of ID8 implantation on muscle mass, fat mass, spleen mass, GDF15 and gene expression of inflammation and atrogenes. Analysis of muscle mass at all time points in hindlimb muscles was completed (A, n = 22–24; soleus (SOL), extensor digitorum longus (EDL), plantaris (PLA), tibialis anterior (TA), gastrocnemius (GA) and quadriceps (QUAD)). Subcutaneous adipose mass in the inguinal fat depot (B, n = 9–12), serum GDF15 (C, n = 8–11) and spleen mass (D, n = 21–22) were also analyzed. mRNA content of inflammatory and atrophy markers interleukin-6 (IL-6), tumour necrosis factor – alpha (TNF-α), atrogin and muscle RING-finger protein-1 (MURF-1) were measured using quantitative PCR in the TA and diaphragm of all groups (E-H, n = 6–8). Results represent mean ± SD. Lettering denotes statistical significance when different from each other ( p < 0.05). C57BL/6J female mice ∼75 days post PBS injection as controls (CTRL); C57BL/6J female mice ∼45 days post ovarian cancer injection (45 Days); C57BL/6J female mice ∼75 days post ovarian cancer injection (75 Days); C57BL/6J female mice ∼90 days post ovarian cancer injection (90 Days). All data was analyzed using a one-way ANOVA or Kruskal–Wallis test when data did not fit normality. All ANOVAs were followed by a two-stage step-up method of Benjamini, Krieger and Yukutieli multiple comparisons test.

Article Snippet: GDF15 (Growth differentiation factor 15) levels were analyzed in serum using the mouse GDF-15 DuoSet ELISA kit according to the manufacturer's instructions (R&D Systems DY6385).

Techniques: Expressing, Muscles, Real-time Polymerase Chain Reaction, Injection

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

Journal: bioRxiv

Article Title: Fetally-encoded GDF15 and maternal GDF15 sensitivity are major determinants of nausea and vomiting in human pregnancy

doi: 10.1101/2023.06.02.542661

Figure Lengend Snippet: A: Dot and box plots illustrating the distribution of circulating GDF15 levels in women of ~15 weeks’ gestation with a history of vomiting in pregnancy vs those reporting no nausea and vomiting in pregnancy. P-value is from an unadjusted linear regression model using natural-log transformed GDF15 concentrations. B: Dot and box plots illustrating the distribution of GDF15 levels (Mean gestational age ~ 10 weeks) in women presenting with hyperemesis gravidarum (HG) and those with low levels of nausea and vomiting in pregnancy. P-value is from an unadjusted linear regression model using natural-log transformed GDF15 concentrations. C: Scatter plot illustrating the relationship between gestational age and GDF15 in the first trimester. The trend lines show predicted values of GDF15 levels (+/− 95% CI) in women with and without HG from a linear regression model of natural-log transformed GDF15 with gestational age and HG status included as predictor variables. The P-values are derived from the same regression model for the effect of HG (HG vs Con) and gestational age (Gest. Age). 5 participants (HG = 1, Control = 4) included in the analysis in panel (B) are not plotted or included in this model as they were recruited after the first trimester.

Article Snippet: To perform Mendelian Randomization between circulating GDF15 levels with HG risk, we harmonized Roche-based GDF15 pQTL, GWAS, and LD reference panels to obtain valid estimates.

Techniques: Transformation Assay, Derivative Assay, Control

Journal: bioRxiv

Article Title: Fetally-encoded GDF15 and maternal GDF15 sensitivity are major determinants of nausea and vomiting in human pregnancy

doi: 10.1101/2023.06.02.542661

Figure Lengend Snippet: A : Schema of experimental design. The GDF15 dimer for maternal and fetal GDF15 is extracted and then digested with the endopeptidase GluC, cutting the N-terminal region into two distinct peptides with glutamic acid C-termini. The stoichiometry of the H and D peptides can then be monitored using LC-MS/MS to determine the relative levels of maternal or fetal derived GDF15 in the maternal circulation. B : Representative LC-MS retention time of H and D peptides from maternal plasma where the mother is heterozygous at H202D and the fetus is homozygous for the H or D allele as indicated. C-E: Scatter plots of the relative quantitation of H peptide vs the D peptide in plasma from pregnancies with the indicated genotypes. The dashed coloured lines in (C-E) indicate the expected relationships between the H and D peptides for the given circulatory origins of GDF15. C: N=20 samples from 5 pregnancies, D : N=8 samples from 2 pregnancies, E : N = 47 samples from 12 pregnancies.

Article Snippet: To perform Mendelian Randomization between circulating GDF15 levels with HG risk, we harmonized Roche-based GDF15 pQTL, GWAS, and LD reference panels to obtain valid estimates.

Techniques: Liquid Chromatography with Mass Spectroscopy, Derivative Assay, Quantitation Assay

Journal: bioRxiv

Article Title: Fetally-encoded GDF15 and maternal GDF15 sensitivity are major determinants of nausea and vomiting in human pregnancy

doi: 10.1101/2023.06.02.542661

Figure Lengend Snippet: A: We studied the secretion of a rare hyperemesis gravidarum risk variant GDF15 C211G. C211G impairs the secretion of GDF15 as determined by western blotting of cell culture medium of cells expressing Flag-tagged wild-type GDF15 (WT-Flag) or GDF15 C211G (C211G-Flag). B: GDF15 C211G impairs the secretion of wild-type GDF15 in a dominant negative manner as co-expression of the mutant inhibited secretion of wild-type GDF15 from 293T cells co-transfected with different amounts (shown in nanograms) of wild-type WT-Flag and Myc-tagged GDF15 C211G (C211G-Myc), as indicated. For A and B representative images from 3 independent experiments are presented. EV represents transfection with the empty plasmid backbone. C: Dot and box plots showing GDF15 levels measured using the Ansh Total GDF15 assay in carriers of GDF15 C211G variant (N=10) identified in an exome-sequencing study of a Croatian population and age and sex matched controls (N=60) derived from the same study. The P-value is from a linear regression model of natural log transformed GDF15 ~ C211G status. D: Forest plot illustrating the effect (standardised betas) of previously described HG risk SNPS on circulating GDF15 measured in 18,184 participants in the Generation Scotland Study. The effect estimates for the rs1054221 variant presented are from an analysis conditioned on the the lead HG variant rs45543339. The beta estimates for GDF15 represent the effect of the HG risk allele on circulating GDF15 in standard deviations. The beta estimates for HG (Hyperemesis Gravidarum) represent the effect of the SNP on risk of HG in log-odds. E: Scatterplot of HG GWAS effect estimates (ie log-odds) vs Roche-based GDF15 pQTL effect estimates derived from cis-Mendelian randomization at the GDF15 locus. MR was performed using m=259 SNPs with genome-wide evidence of pQTL effects on GDF15 levels within 1Mb GDF15 locus and adjusted using LD estimates from UK Biobank WGS individuals (n=138335; see ). Causal effect estimates obtained using LD-aware MR and reflected as regression lines.

Article Snippet: To perform Mendelian Randomization between circulating GDF15 levels with HG risk, we harmonized Roche-based GDF15 pQTL, GWAS, and LD reference panels to obtain valid estimates.

Techniques: Variant Assay, Western Blot, Cell Culture, Expressing, Dominant Negative Mutation, Mutagenesis, Transfection, Plasmid Preparation, Sequencing, Derivative Assay, Transformation Assay, Genome Wide

Journal: bioRxiv

Article Title: Fetally-encoded GDF15 and maternal GDF15 sensitivity are major determinants of nausea and vomiting in human pregnancy

doi: 10.1101/2023.06.02.542661

Figure Lengend Snippet: A : Schema of the experimental paradigm. Single housed, adult male and female C57Bl/6J mice were injected with 0.01mg/kg of Fc-GDF15–15 fusion protein (Fc_GDF15) or vehicle control (PBS). Food intake was measured overnight (from 5pm to 9am) before (black bar) and after treatment (red bar) with standard, short acting human recombinant GDF-15 (hrGDF15 0.1mg/kg). Body weight was measured at the same timepoints. B : Food intake recorded overnight (5pm-9am) the day before (black dots) and after an acute bolus of hrGDF15 (red dots) in mice with and without pre-treatment with Fc_GDF15. C: Body weight at 9am the day before (black dots) and 9 am the day after (red dots) an acute bolus of hrGDF15 in mice with and without pre-treatment with Fc_GDF15. N=17 (12 male, 5 female) in Control and 19 in FC_GDF15 group (13 male, 6 female). Data were analysed using mixed-effect analysis, post-hoc testing comparing before and after acute GDF-15 treatment was undertaken with the Sidak test to correct for multiple testing. *P<0.05, ***P<0.001, ****P<0.0001, ns = non-significant, P>0.05.

Article Snippet: To perform Mendelian Randomization between circulating GDF15 levels with HG risk, we harmonized Roche-based GDF15 pQTL, GWAS, and LD reference panels to obtain valid estimates.

Techniques: Injection, Control, Recombinant