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Circulating biomarkers levels in chronic heart failure patients and controls

Journal: ESC Heart Failure

Article Title: Integrated miRNA-cytokine profiling reveals a molecular signature of cachexia in patients with chronic heart failure

doi: 10.1093/eschf/xvag229

Figure Lengend Snippet: Circulating biomarkers levels in chronic heart failure patients and controls

Article Snippet: Serum concentrations of GDF-15 (#ELH-GDF15-1) and FGF-21 (#ELH-FGF21-1) were measured using ELISA (RayBiotech, Norcross, GA, USA) according to manufacturers’ instructions.

Techniques:

Circulating miRNAs and cytokines as molecular signatures of heart failure. Differences in serum levels of GDF-15 and FGF-21 in CHF patients and controls. Serum levels of GDF-15 by ELISA assay in patients with heart failure (HF) ( n = 25) and the control group ( n = 10). ( A ) Data showed significant upregulation of GDF-15 in CHF vs controls ( P < .001). ( B ) FGF-21 levels did not differ significantly between groups. Correlations between GDF-15 and selected miRNAs in CHF patients. ( C , D ) In CHF patients, GDF-15 levels were positively associated with miR-29b ( ρ = 0.42, P = .037) and miR-486 ( ρ = 0.42, P = .039). Data are expressed as fold change and are presented as mean ± SEM. Student's t -test or Mann–Whitney test was used for normally distributed data or non-normally distributed data, respectively. *** P ≤ .001.

Journal: ESC Heart Failure

Article Title: Integrated miRNA-cytokine profiling reveals a molecular signature of cachexia in patients with chronic heart failure

doi: 10.1093/eschf/xvag229

Figure Lengend Snippet: Circulating miRNAs and cytokines as molecular signatures of heart failure. Differences in serum levels of GDF-15 and FGF-21 in CHF patients and controls. Serum levels of GDF-15 by ELISA assay in patients with heart failure (HF) ( n = 25) and the control group ( n = 10). ( A ) Data showed significant upregulation of GDF-15 in CHF vs controls ( P < .001). ( B ) FGF-21 levels did not differ significantly between groups. Correlations between GDF-15 and selected miRNAs in CHF patients. ( C , D ) In CHF patients, GDF-15 levels were positively associated with miR-29b ( ρ = 0.42, P = .037) and miR-486 ( ρ = 0.42, P = .039). Data are expressed as fold change and are presented as mean ± SEM. Student's t -test or Mann–Whitney test was used for normally distributed data or non-normally distributed data, respectively. *** P ≤ .001.

Article Snippet: Serum concentrations of GDF-15 (#ELH-GDF15-1) and FGF-21 (#ELH-FGF21-1) were measured using ELISA (RayBiotech, Norcross, GA, USA) according to manufacturers’ instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Control, MANN-WHITNEY

Molecular profiles in CHF patients. Differences in serum levels of GDF-15 and FGF-21 among heart failure patients with and without cardiac cachexia. Circulating levels of GDF-15 and FGF-21 were analyzed in CHF patients with cardiac cachexia (CC) ( n = 10) and without cardiac cachexia (nCC) ( n = 15), and a control group ( n = 10). ( A ) Significant difference in GDF-15 levels was observed between patients with CC or nCC vs controls (all P < .001). ( B ) Levels of FGF-21 were significantly higher in CHF with CC compared with nCC ( P = .012) and vs controls ( P = .010) ( C ). Spearman’s correlation analysis revealed that in CHF patients with CC, GDF-15 negatively correlated with miR-21 ( ρ = −0.81, P = .007). ( D ) In the same group, FGF-21 negatively correlated with miR-486 ( ρ = −0.69, P = .035). Data are expressed as fold change and are presented as mean ± SEM. One-way ANOVA test or Kruskal–Wallis test was used for normally distributed data or non-normally distributed data, respectively. * P < .05, *** P ≤ .001.

Journal: ESC Heart Failure

Article Title: Integrated miRNA-cytokine profiling reveals a molecular signature of cachexia in patients with chronic heart failure

doi: 10.1093/eschf/xvag229

Figure Lengend Snippet: Molecular profiles in CHF patients. Differences in serum levels of GDF-15 and FGF-21 among heart failure patients with and without cardiac cachexia. Circulating levels of GDF-15 and FGF-21 were analyzed in CHF patients with cardiac cachexia (CC) ( n = 10) and without cardiac cachexia (nCC) ( n = 15), and a control group ( n = 10). ( A ) Significant difference in GDF-15 levels was observed between patients with CC or nCC vs controls (all P < .001). ( B ) Levels of FGF-21 were significantly higher in CHF with CC compared with nCC ( P = .012) and vs controls ( P = .010) ( C ). Spearman’s correlation analysis revealed that in CHF patients with CC, GDF-15 negatively correlated with miR-21 ( ρ = −0.81, P = .007). ( D ) In the same group, FGF-21 negatively correlated with miR-486 ( ρ = −0.69, P = .035). Data are expressed as fold change and are presented as mean ± SEM. One-way ANOVA test or Kruskal–Wallis test was used for normally distributed data or non-normally distributed data, respectively. * P < .05, *** P ≤ .001.

Article Snippet: Serum concentrations of GDF-15 (#ELH-GDF15-1) and FGF-21 (#ELH-FGF21-1) were measured using ELISA (RayBiotech, Norcross, GA, USA) according to manufacturers’ instructions.

Techniques: Control, Analysis

Molecular profiles in cachectic patients with severe CHF. Circulating miRNA levels and GDF-15 correlations according to ejection fraction and cachexia status in CHF patients. Circulating miRNA levels were analyzed in CHF patients with preserved ejection fraction without cachexia (HFpEF + nCC) ( n = 7) and with reduced ejection fraction with cachexia (HFrEF + CC) ( n = 5) and a control group ( n = 10). ( A ) Data shows significant downregulation of miR-29b ( P = .019) in HFrEF + CC vs HFpEF + nCC and vs controls ( P = .002). ( B ) In HFpEF + nCC, GDF-15 showed a strong positive correlation with miR-29b ( r = 0.82, P = .034). Data are expressed as fold change and are presented as mean ± SEM. One-way ANOVA test or Kruskal–Wallis test was used for normally distributed data or non-normally distributed data, respectively. * P < .05, ** P ≤ .01.

Journal: ESC Heart Failure

Article Title: Integrated miRNA-cytokine profiling reveals a molecular signature of cachexia in patients with chronic heart failure

doi: 10.1093/eschf/xvag229

Figure Lengend Snippet: Molecular profiles in cachectic patients with severe CHF. Circulating miRNA levels and GDF-15 correlations according to ejection fraction and cachexia status in CHF patients. Circulating miRNA levels were analyzed in CHF patients with preserved ejection fraction without cachexia (HFpEF + nCC) ( n = 7) and with reduced ejection fraction with cachexia (HFrEF + CC) ( n = 5) and a control group ( n = 10). ( A ) Data shows significant downregulation of miR-29b ( P = .019) in HFrEF + CC vs HFpEF + nCC and vs controls ( P = .002). ( B ) In HFpEF + nCC, GDF-15 showed a strong positive correlation with miR-29b ( r = 0.82, P = .034). Data are expressed as fold change and are presented as mean ± SEM. One-way ANOVA test or Kruskal–Wallis test was used for normally distributed data or non-normally distributed data, respectively. * P < .05, ** P ≤ .01.

Article Snippet: Serum concentrations of GDF-15 (#ELH-GDF15-1) and FGF-21 (#ELH-FGF21-1) were measured using ELISA (RayBiotech, Norcross, GA, USA) according to manufacturers’ instructions.

Techniques: Control

Schematic representation of the circulating molecular profile associated with chronic heart failure and cardiac cachexia, characterized by dysregulation of muscle- and remodelling-related miRNAs and increased stress-responsive cytokines. CHF patients showed reduced miR-15b, miR-29b, miR-133a, and miR-486 and increased GDF-15, while cardiac cachexia was further associated with lower miR-29b and higher FGF-21 levels.

Journal: ESC Heart Failure

Article Title: Integrated miRNA-cytokine profiling reveals a molecular signature of cachexia in patients with chronic heart failure

doi: 10.1093/eschf/xvag229

Figure Lengend Snippet: Schematic representation of the circulating molecular profile associated with chronic heart failure and cardiac cachexia, characterized by dysregulation of muscle- and remodelling-related miRNAs and increased stress-responsive cytokines. CHF patients showed reduced miR-15b, miR-29b, miR-133a, and miR-486 and increased GDF-15, while cardiac cachexia was further associated with lower miR-29b and higher FGF-21 levels.

Article Snippet: Serum concentrations of GDF-15 (#ELH-GDF15-1) and FGF-21 (#ELH-FGF21-1) were measured using ELISA (RayBiotech, Norcross, GA, USA) according to manufacturers’ instructions.

Techniques:

(A) Proteome Profiler Human XL cytokine array (105 cytokines) of conditioned medium from proliferating, doxorubicin-induced (Dox), and Gem/Cis-induced senescent-like RBE cells. (B) Gdf15 mRNA in proliferating versus senescent-like murine CCA (SB1) cells. (C) Gdf15 mRNA in control vs AP-treated SB-p16-ATTAC tumors. (D) Schematic of in vitro studies. (E-F) Arg1 and iNOS mRNA in bone-marrow-derived macrophage (BMDM) incubated for 72 h with conditioned medium from vehicle- or alisertib-treated SB1 cells expressing shCtrl or shGdf15 . (G) Bulk RNA-seq heatmap (row Z-scores) of naïve bone marrow-derived macrophages (BMDM) cultured 72 h with conditioned media (CM) from SB-p16-shCtrl or SB- p16-Gdf15 OE cells. (H) ARG1 mRNA in human monocyte-derived macrophages ± recombinant human GDF-15 (Qkine Qk017; TGF-β1-free), qRT-PCR relative to vehicle; mean ± SEM, n = 6. (I) Division index (CFSE dilution, normalized to T-cells-only) of anti-CD3/CD28-activated CD8⁺ T cells co-cultured with BMDMs incubated for 72 h with conditioned medium from vehicle- or alisertib-treated SB1 cells expressing shCtrl or shGdf15 . Data are mean ± SEM; *, P<0.05; **, P<0.01; ****, P<0.0001; ns, not significant.

Journal: bioRxiv

Article Title: Therapy-induced senescent-like cancer cells drive macrophage-mediated immunosuppression in cholangiocarcinoma

doi: 10.64898/2026.06.24.734341

Figure Lengend Snippet: (A) Proteome Profiler Human XL cytokine array (105 cytokines) of conditioned medium from proliferating, doxorubicin-induced (Dox), and Gem/Cis-induced senescent-like RBE cells. (B) Gdf15 mRNA in proliferating versus senescent-like murine CCA (SB1) cells. (C) Gdf15 mRNA in control vs AP-treated SB-p16-ATTAC tumors. (D) Schematic of in vitro studies. (E-F) Arg1 and iNOS mRNA in bone-marrow-derived macrophage (BMDM) incubated for 72 h with conditioned medium from vehicle- or alisertib-treated SB1 cells expressing shCtrl or shGdf15 . (G) Bulk RNA-seq heatmap (row Z-scores) of naïve bone marrow-derived macrophages (BMDM) cultured 72 h with conditioned media (CM) from SB-p16-shCtrl or SB- p16-Gdf15 OE cells. (H) ARG1 mRNA in human monocyte-derived macrophages ± recombinant human GDF-15 (Qkine Qk017; TGF-β1-free), qRT-PCR relative to vehicle; mean ± SEM, n = 6. (I) Division index (CFSE dilution, normalized to T-cells-only) of anti-CD3/CD28-activated CD8⁺ T cells co-cultured with BMDMs incubated for 72 h with conditioned medium from vehicle- or alisertib-treated SB1 cells expressing shCtrl or shGdf15 . Data are mean ± SEM; *, P<0.05; **, P<0.01; ****, P<0.0001; ns, not significant.

Article Snippet: Human recombinant GDF-15 was from Qkine (Qk017) and murine recombinant GDF-15 from R&D Systems (8944-GD/CF).

Techniques: Control, In Vitro, Derivative Assay, Incubation, Expressing, RNA Sequencing, Cell Culture, Recombinant, Quantitative RT-PCR

(A) Macrophages express TGFBR2, the candidate non-canonical GDF-15 receptor, in murine and human CCA single-cell RNA-seq. (B) α-TGFBR2 western blot (WB) of streptavidin pull-down (top) and input (middle) from naïve BMDMs treated with biotin-supplemented conditioned media (CM) from CCA cells expressing GDF-15-BioID2-HA or SEC61B-BioID2-HA (ER control); α-HA blot of CM (bottom) confirms bait secretion. (C) GDF-15 induces macrophage phospho-STAT6 but not phospho-SMAD2/3, reflecting non-canonical TGFBR2 output. (D) WB of BMDMs stimulated with control or GDF-15-overexpressing (GDF-15 OE ) CM for 0 or 1 h: p-STAT6, STAT6, p-SMAD3, SMAD3. (E) Arg1 mRNA in BMDMs transfected with siCtrl or siTgfbr2 and stimulated with control or rGDF-15. n = 6. Data are mean ± SEM; *, P<0.05; **, P<0.01; ****, P<0.0001; ns, not significant.

Journal: bioRxiv

Article Title: Therapy-induced senescent-like cancer cells drive macrophage-mediated immunosuppression in cholangiocarcinoma

doi: 10.64898/2026.06.24.734341

Figure Lengend Snippet: (A) Macrophages express TGFBR2, the candidate non-canonical GDF-15 receptor, in murine and human CCA single-cell RNA-seq. (B) α-TGFBR2 western blot (WB) of streptavidin pull-down (top) and input (middle) from naïve BMDMs treated with biotin-supplemented conditioned media (CM) from CCA cells expressing GDF-15-BioID2-HA or SEC61B-BioID2-HA (ER control); α-HA blot of CM (bottom) confirms bait secretion. (C) GDF-15 induces macrophage phospho-STAT6 but not phospho-SMAD2/3, reflecting non-canonical TGFBR2 output. (D) WB of BMDMs stimulated with control or GDF-15-overexpressing (GDF-15 OE ) CM for 0 or 1 h: p-STAT6, STAT6, p-SMAD3, SMAD3. (E) Arg1 mRNA in BMDMs transfected with siCtrl or siTgfbr2 and stimulated with control or rGDF-15. n = 6. Data are mean ± SEM; *, P<0.05; **, P<0.01; ****, P<0.0001; ns, not significant.

Article Snippet: Human recombinant GDF-15 was from Qkine (Qk017) and murine recombinant GDF-15 from R&D Systems (8944-GD/CF).

Techniques: Single Cell, RNA Sequencing, Western Blot, Expressing, Control, Transfection

(A) Strategy for p16-promoter-restricted Gdf15 silencing (shRNA) and Gdf15 overexpression. (B) qPCR validation of Gdf15 knockdown and overexpression in senescent-like SB1 cells. (C-D) Subcutaneous model and tumor-burden quantification. (E) GeoMx spatial profiling of GDF-15 expression and macrophage infiltration in human CCA. Region-of-interest (ROI)-level analysis plotting GDF-15 expression against macrophage score; each point is one ROI, with the linear fit and 95% confidence band shown. Spearman ρ = 0.63, P = 0.0067. Data are mean ± SEM; **, P<0.01; ****, P<0.0001.

Journal: bioRxiv

Article Title: Therapy-induced senescent-like cancer cells drive macrophage-mediated immunosuppression in cholangiocarcinoma

doi: 10.64898/2026.06.24.734341

Figure Lengend Snippet: (A) Strategy for p16-promoter-restricted Gdf15 silencing (shRNA) and Gdf15 overexpression. (B) qPCR validation of Gdf15 knockdown and overexpression in senescent-like SB1 cells. (C-D) Subcutaneous model and tumor-burden quantification. (E) GeoMx spatial profiling of GDF-15 expression and macrophage infiltration in human CCA. Region-of-interest (ROI)-level analysis plotting GDF-15 expression against macrophage score; each point is one ROI, with the linear fit and 95% confidence band shown. Spearman ρ = 0.63, P = 0.0067. Data are mean ± SEM; **, P<0.01; ****, P<0.0001.

Article Snippet: Human recombinant GDF-15 was from Qkine (Qk017) and murine recombinant GDF-15 from R&D Systems (8944-GD/CF).

Techniques: shRNA, Over Expression, Biomarker Discovery, Knockdown, Expressing