Journal: Cardiology Discovery

Article Title: Cardioprotective Effect of Growth Differentiation Factor 15 Against Isoproterenol-Induced Cardiomyocyte Apoptosis via Regulation of the Mitochondrial Fusion

doi: 10.1097/cd9.0000000000000051

Figure Lengend Snippet: Figure 1: GDF15 is highly expressed in the serum and heart tissue of patients with HF. (A) Serum GDF15 levels in HF group (n=57) and the non-HF group (n=57). (B) Correlation analysis between GDF15 concentration and NYHA class (n=114). (C) Immunohistochemical analysis demonstrated higher GDF15 levels in myocardial tissue obtained from HF group compared to non-HF group. GDF15: Growth differentiation factor 15; HF: Heart failure; NYHA: New York Heart Association.

Article Snippet: Human or mouse GDF15 concentrations were determined in serum in single measurements using a quantitative sandwich monoclonal enzyme-linked immunosorbent assay (DGD150, R&D System, Inc., Minneapolis, Minnesota, USA).

Techniques: Concentration Assay, Immunohistochemical staining

Journal: Cardiology Discovery

Article Title: Cardioprotective Effect of Growth Differentiation Factor 15 Against Isoproterenol-Induced Cardiomyocyte Apoptosis via Regulation of the Mitochondrial Fusion

doi: 10.1097/cd9.0000000000000051

Figure Lengend Snippet: Figure 2: GDF15 expression in the ISO-induced HF mouse model. HF group treated with ISO (30mg/(kg·day)) for 7days (n=6) or 28days (n=6), non-HF group received saline for 28days (n=5). (A and B) EF and FS values in mice treated with ISO for 7 and 28days or non-HF group for 28days. (C) Serum GDF15 levels in ISO-induced mice after ISO treatment for 7 and 28days or non-HF group for 28days. (D) Western blotting for GDF15 expression in heart tissue from ISO-treated mice for 7 and 28days or non-HF group for 28days. (E) HE, Masson staining, and GDF15 expression in heart tissue with or without ISO administration. ∗P<0.01, †P<0.001 versus non-HF group. EF: Ejection fraction; FS: Fractional shortening; GADPH: Glyceraldehyde-3-phosphate dehydrogenase; GDF15: Growth differentiation factor 15; HE: Hematoxylin-eosin; HF: Heart failure; ISO: Isoproterenol.

Article Snippet: Human or mouse GDF15 concentrations were determined in serum in single measurements using a quantitative sandwich monoclonal enzyme-linked immunosorbent assay (DGD150, R&D System, Inc., Minneapolis, Minnesota, USA).

Techniques: Expressing, Saline, Western Blot, Staining

Journal: Cardiology Discovery

Article Title: Cardioprotective Effect of Growth Differentiation Factor 15 Against Isoproterenol-Induced Cardiomyocyte Apoptosis via Regulation of the Mitochondrial Fusion

doi: 10.1097/cd9.0000000000000051

Figure Lengend Snippet: Figure 3: ISO increased GDF15 levels in CMs, and this was associated with oxidative inflammation and apoptosis. (A and B) Representative Western blotting and quantification for GDF15, COX2, and cleaved caspase 3 in ISO (0.5mmol/L)-treated CMs over time. (C) GDF15 mRNA levels in ISO (0.5mmol/L)-treated CMs over time. (D and E) Representative Western blotting and quantification for GDF15, COX2, and cleaved caspase 3 in ISO (0.5mmol/L)-treated primary CMs. (F and G) TUNEL staining and quantification analysis for CMs apoptosis with or without ISO (0.5mmol/L) treatment for 24 and 48hours. Differences were determined by 2- sided Student’s t tests. ∗P<0.05, †P<0.01, ‡P<0.001 versus control group. CM: Cardiomyocytes; COX2: Cyclooxygenase 2; GADPH: Glyceraldehyde-3- phosphate dehydrogenase; GDF15: Growth differentiation factor 15; ISO: Isoproterenol.

Article Snippet: Human or mouse GDF15 concentrations were determined in serum in single measurements using a quantitative sandwich monoclonal enzyme-linked immunosorbent assay (DGD150, R&D System, Inc., Minneapolis, Minnesota, USA).

Techniques: Western Blot, TUNEL Assay, Staining, Control

Journal: Cardiology Discovery

Article Title: Cardioprotective Effect of Growth Differentiation Factor 15 Against Isoproterenol-Induced Cardiomyocyte Apoptosis via Regulation of the Mitochondrial Fusion

doi: 10.1097/cd9.0000000000000051

Figure Lengend Snippet: Figure 4: Silencing GDF15 exaggerated ISO-induced CM damage. (A) GDF15 mRNA levels in cells transfected with GDF15 siRNA. (B and C) Western blotting and quantification for GDF15 in GDF15 siRNA-transfected cells. (D) Ratio of apoptosis in ISO-treated cells with or without GDF silencing for 24hours. (E–H) Representative blotting and quantification for GDF15, COX2, and cleaved caspase 3 in ISO-treated cells with or without GDF15 silencing for 24hours. (I–L) Representative blotting and quantification for GDF15, COX2, and cleaved caspase 3 in ISO-treated cells with or without pcDNA3.1-GDF15 transfection for 24 hours. Differences were determined by 2-sided Student’s t tests. ∗P<0.01, †P<0.001 versus control group. ‡P<0.05 versus siControl group. CMs: Cardiomyocytes; COX2: Cyclooxygenase-2; GADPH: Glyceraldehyde-3-phosphate dehydrogenase; GDF15: Growth differentiation factor 15; ISO: Isoproterenol; siControl: siRNA for control; siGDF15: siRNA for GDF15.

Article Snippet: Human or mouse GDF15 concentrations were determined in serum in single measurements using a quantitative sandwich monoclonal enzyme-linked immunosorbent assay (DGD150, R&D System, Inc., Minneapolis, Minnesota, USA).

Techniques: Transfection, Western Blot, Control

Journal: Cardiology Discovery

Article Title: Cardioprotective Effect of Growth Differentiation Factor 15 Against Isoproterenol-Induced Cardiomyocyte Apoptosis via Regulation of the Mitochondrial Fusion

doi: 10.1097/cd9.0000000000000051

Figure Lengend Snippet: Figure 5: GDF15 regulated the balance of mitochondrial fission and fusion. (A–C) Western blotting and quantification for mitochondrial marker proteins MFN2, OPA-1, FIS-1, MFF, and DRP-1 in CMs treated with ISO for 0, 24, and 48hours. (D) MitoTracker staining of CMs with or without ISO treatment. (E–J) Expression and quantification of mitochondrial marker proteins MFN2, OPA-1, FIS-1, MFF, and DRP-1 in ISO-treated CMs with or without GDF15 silencing. (K) MitoTracker and GDF15 co-staining in CMs with or without GDF15 silencing. Differences were determined by 2-sided Student’s t tests. ∗P<0.01, †P<0.001 versus siControl group without ISO treatment or 0 hour. ‡P<0.05 versus siControl group without ISO treatment or 0 hour. CMs: Cardiomyocytes; DRP-1: Dynamin-related protein-1; FIS-1: Fission 1 protein; GADPH: Glyceraldehyde-3-phosphate dehydrogenase; GDF15: Growth differentiation factor 15; ISO: Isoproterenol; MFF: Mitochondrial fission factor; MFN2: Mitofusin 2; OPA-1: Optic atrophy 1; siControl: siRNA for control; siGDF15: siRNA for GDF15.

Article Snippet: Human or mouse GDF15 concentrations were determined in serum in single measurements using a quantitative sandwich monoclonal enzyme-linked immunosorbent assay (DGD150, R&D System, Inc., Minneapolis, Minnesota, USA).

Techniques: Western Blot, Marker, Staining, Expressing, Control

Journal: Journal for immunotherapy of cancer

Article Title: Loss of ADAR1 in macrophages in combination with interferon gamma suppresses tumor growth by remodeling the tumor microenvironment.

doi: 10.1136/jitc-2023-007402

Figure Lengend Snippet: Figure 3 ADAR1 loss in macrophage with IFN-γ treatment affects the secretion of key cytokines through PKR/EIF2α signaling. (A, C) Human XL cytokine arrays for detecting differential factors between THP-1 cells with scrambled shRNA and shADAR1#1 under the treatment of IFN-γ (A) and between THP-1 cells with empty vector and WT ADAR, co-cultured with A549 (C). (B, D) Bar plots showing the expression levels of differential factors on ADAR1 knockdown (B) and ADAR1 overexpression (D). (E) Venn gram showing key cytokines identified by ADAR1 knockdown and overexpression experiments and their potential effects on the tumor microenvironment. (F) RT-qPCR-based mRNA expression levels of ADAR, CCL20, GDF15, IFN-G, IL-18, IL-18BP, and HAVCR2 in different THP-1 cells (transfected with scrambled shRNA, shADAR1#1 or shADAR1#2) with IFN-γ treatment. β-actin was used as an internal control. (G) RT-qPCR-based mRNA expression levels of Ccl20, Gdf15, Il-18, Il-18bp, and Havcr2 in BMDMs from C57BL/6 mice (Adarfl/fl and Adarfl/flLyz2Cre) treated with IFN-γ. Gapdh was used as an internal control. (H) Immunofluorescent staining for anti-dsRNA (J2) in THP-1 cells treated with IFN-γ. RNase III treatment was used as the negative control for the dsRNA signal. Scale bars, 10 µm. (I) Western blot showing the protein expression of p-PKRThr446/

Article Snippet: 2- AP (GlpBio, GC61906), Phorbol 12- myristate 13- acetate (PMA) (MedChemExpress, HY- 18739), Recombinant Human IFN-γ (Novoprotein, C014), Recombinant Mouse IL- 2 (Novoprotein, P04351), Recombinant M- CSF (Novoprotein, CB34), Recombinant Mouse IFN-γ (Novoprotein, C746), Recombinant Mouse TIM- 3 (Novoprotein, CM54), Recombinant Mouse CCL20 (PeproTech, 250–2), Recombinant Mouse GDF15 (R&D Systems, 8944- GD025), Recombinant Mouse IL- 18 Binding Protein Isoform d (Novoprotein, CM45), ShortCut RNase III (NEB, M0245S), poly (I:C) (GlpBio, GC14710).

Techniques: shRNA, Plasmid Preparation, Cell Culture, Expressing, Knockdown, Over Expression, Quantitative RT-PCR, Transfection, Control, Staining, Negative Control, Western Blot

Journal: Journal for immunotherapy of cancer

Article Title: Loss of ADAR1 in macrophages in combination with interferon gamma suppresses tumor growth by remodeling the tumor microenvironment.

doi: 10.1136/jitc-2023-007402

Figure Lengend Snippet: Figure 5 The effect of ADAR1 loss and key cytokines on tube formation. (A) Schematic diagram of tube formation experiment of SVEC4-10 cells. (B) Tube formation of SVEC4-10 cells treated with conditioned media from BMDMs (Adarfl/fl vs Adarfl/flLyz2Cre) and CCL20, GDF15, or IFN-γ. Scale bar, 500 µm. (C) Bar plots showing relative vessel area, the total number of junctions, and the vessel length for SVEC4-10 tube formation. (D) Tube formation of SVEC4-10 cells treated with conditioned media from BMDMs (Adarfl/flLyz2Cre) and IFN-γ after pretreatment with or without 2-AP (5 mM). Scale bar, 500 µm. (E) Bar plots showing relative vessel area, the total number of junctions, and vessel length for SVEC4-10 tube formation. (F) Tube formation of HUVEC cells treated with conditioned media from THP-1 cells (transfected with shADAR1#1) and IFN-γ after pretreatment with or without 2-AP (5 mM). Scale bar, 200 µm. (G) Bar plots showing relative vessel area, the total number of junctions, and the vessel length for HUVEC tube formation. (C, E, and G) P values are based on unpaired Student’s t-test. Data are presented as mean±SD. ADAR, adenosine deaminases acting on RNA; BMDMs, bone marrow-derived macrophages; IFN, interferon; 2-AP, 2-aminopurine.

Article Snippet: 2- AP (GlpBio, GC61906), Phorbol 12- myristate 13- acetate (PMA) (MedChemExpress, HY- 18739), Recombinant Human IFN-γ (Novoprotein, C014), Recombinant Mouse IL- 2 (Novoprotein, P04351), Recombinant M- CSF (Novoprotein, CB34), Recombinant Mouse IFN-γ (Novoprotein, C746), Recombinant Mouse TIM- 3 (Novoprotein, CM54), Recombinant Mouse CCL20 (PeproTech, 250–2), Recombinant Mouse GDF15 (R&D Systems, 8944- GD025), Recombinant Mouse IL- 18 Binding Protein Isoform d (Novoprotein, CM45), ShortCut RNase III (NEB, M0245S), poly (I:C) (GlpBio, GC14710).

Techniques: Transfection, Derivative Assay

Journal: Journal for immunotherapy of cancer

Article Title: Loss of ADAR1 in macrophages in combination with interferon gamma suppresses tumor growth by remodeling the tumor microenvironment.

doi: 10.1136/jitc-2023-007402

Figure Lengend Snippet: Figure 8 Schematic summary of this study. ADAR1-deficient macrophages combined with IFN-γ treatment reprogram the tumor microenvironment by two mechanisms: (1) inhibit angiogenesis by decreased secretion of GDF15 and CCL20 and increased secretion of IFN-γ, and (2) activate CD8+ T cells by decreased secretion of TIM-3 and IL-18BP and increased secretion of IL-18. These effects collectively convert a “cold tumor” into a “hot tumor.” Combined treatment with ADAR1- deficient macrophages and IFN-γ may represent an effective therapeutic approach. dsRNA, double-stranded RNA; IFN, interferon; IL, interleukin; uORF, upstream Open Reading Frame.

Article Snippet: 2- AP (GlpBio, GC61906), Phorbol 12- myristate 13- acetate (PMA) (MedChemExpress, HY- 18739), Recombinant Human IFN-γ (Novoprotein, C014), Recombinant Mouse IL- 2 (Novoprotein, P04351), Recombinant M- CSF (Novoprotein, CB34), Recombinant Mouse IFN-γ (Novoprotein, C746), Recombinant Mouse TIM- 3 (Novoprotein, CM54), Recombinant Mouse CCL20 (PeproTech, 250–2), Recombinant Mouse GDF15 (R&D Systems, 8944- GD025), Recombinant Mouse IL- 18 Binding Protein Isoform d (Novoprotein, CM45), ShortCut RNase III (NEB, M0245S), poly (I:C) (GlpBio, GC14710).

Techniques: