gdf15  (Novus Biologicals)

Journal: Cancer research

Article Title: GDF15 Reprograms the Microenvironment to Drive Liver Metastasis of Uveal Melanoma

doi: 10.1158/0008-5472.CAN-25-0536

Figure Lengend Snippet: A: UMAP of 6 individual clinical UM “and healthy” scRNA-seq datasets. Data shows major cell types identified and then GDF15 expression in the UM cells (malignant cells) B: Violin plots showing expression of GDF15 in a scRNA-Seq analysis of human Class 1 and Class 2 UM. C: Western Blot showing the expression of BAP1, PRAME and GDF15 in a panel of UM cell lines. D: Silencing of BAP1 is associated with increased expression of GDF15 in Mel202 and 92.1 UM cells. BAP1 was silenced using siRNA and resulting lysates probed for expression of BAP1 and GDF15. E: ChIP-Seq analysis of Mel202 cells expressing BAP1 or silenced for BAP1 demonstrates an increase in H3K27ac at the GDF15 promoter following BAP1 knockdown. F: Analysis of GDF15 secretion by ELISA assay demonstrates that co-culture of UM cells with HSCs is associated with increased GDF15 expression, even in UM cell lines with high basal expression of GDF15. Cell lines derived from primary or metastatic specimens are indicated.

Article Snippet: The effectiveness of blocking GDF15 on tube formation was assessed using an anti-GDF15 blocking antibody (ponsegromab: 120ng/ml, #HY-P99241, MedChemExpress, NJ, USA).

Techniques: Expressing, Western Blot, ChIP-sequencing, Knockdown, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Derivative Assay

Journal: Cancer research

Article Title: GDF15 Reprograms the Microenvironment to Drive Liver Metastasis of Uveal Melanoma

doi: 10.1158/0008-5472.CAN-25-0536

Figure Lengend Snippet: A: RNA-seq analysis of HSCs (LX2) treated with GDF15 (1μg/ml, 24 hr) identifies increased expression of genes encoding multiple ECM proteins ( COL7A1 ), and molecules involved in angiogenesis ( VCAM1, ANGPLT4, PDGFB, FLT1 ). B: Pathway analysis identifies GDF15 to increase expression of genes involved in inflammatory signaling ( TNFA, IFNγ, IL6 ), angiogenesis (hypoxia, angiogenesis) and metabolism. C: Human primary HSCs (HHStec) were stimulated with either GDF15 (1μg/ml) or SPP1 (osteopontin, (1μg/ml)) for 24 hr and Western Blotting used to determine expression of IL-8 and phospho-STAT3. D: HSC (HHStec cells) were grown in monoculture, MP41 cells were grown in monoculture, HSC+MP41 cells were grown in co-culture, HSCs were treated with conditioned media (CM) from MP41 cells or GDF15 (1μg/ml) for 24 hr before probing for the expression of COL1A1 and PDGFRB by Western Blot. E: Immunofluorescence staining of COL1A1 in HSCs (HHStec) cells treated with basal media or conditioned media from MP41 or UMM061 cells. F: CellChat analysis of human UM samples identifies HSC-derived collagens as being a potential outgoing signal to multiple immune subtypes and UM cells.

Article Snippet: The effectiveness of blocking GDF15 on tube formation was assessed using an anti-GDF15 blocking antibody (ponsegromab: 120ng/ml, #HY-P99241, MedChemExpress, NJ, USA).

Techniques: Expressing, RNA Sequencing, Western Blot, Co-Culture Assay, Immunofluorescence, Staining, Derivative Assay

Journal: Cancer research

Article Title: GDF15 Reprograms the Microenvironment to Drive Liver Metastasis of Uveal Melanoma

doi: 10.1158/0008-5472.CAN-25-0536

Figure Lengend Snippet: A: CellChat analysis of scRNA-Seq from human UM identifies endothelial cells as being a major target of secreted GDF15. B: Exogenous GDF15 and IL8 increase vascular network formation. HUVEC cells were plated on Matrigel and treated with basal media, IL-8 or GDF15 (both 1μg/ml for 0–24 hr). C: Scoring of endothelial network formation data from B. D: Conditioned media (CM) from UM-HSC co-cultures increases vascular network formation. HUVECs were plated onto Matrigel and treated with basal media or CM from MP41, UMM061 and MM28 cells grown in co-culture with LX2 cells. E: Vascular network formation from D was quantified. F: GDF15-blocking antibodies reverse the pro-angiogenic effects of CM. HUVEC cells were treated with CM from MP41-LX2 co-cultures in the absence and presence of the GDF15 blocking antibody (Ponsegromab,120ng/ml) and vascular network formation imaged. G : Quantification of endothelial cell network formation from F.

Article Snippet: The effectiveness of blocking GDF15 on tube formation was assessed using an anti-GDF15 blocking antibody (ponsegromab: 120ng/ml, #HY-P99241, MedChemExpress, NJ, USA).

Techniques: Co-Culture Assay, Blocking Assay

Journal: Cancer research

Article Title: GDF15 Reprograms the Microenvironment to Drive Liver Metastasis of Uveal Melanoma

doi: 10.1158/0008-5472.CAN-25-0536

Figure Lengend Snippet: A: IVIS imaging of mice with either shCRTL or shGDF15#2 MP41 cells at days 49 and 77, respectively. B: Metastasis-free survival of mice following tail vein injection of either shCRTL or shGDF15#2 MP41 cells. C: H&E staining of livers following tail vein injection of shCRTL or GDF15 silenced (GDF15 shRNA#2) MP41 UM cells. Livers were collected after 7 weeks (shCRTL) or 11 weeks (shGDF15). Scale Bar = 6 mm. D: Quantification of liver metastases following tail vein injection of GDF15-expressing or silenced MP41 cells (GDF15 shRNA#1 and shRNA#2). E: IVIS imaging of mice with either shCRTL or shGDF15#2 OMM1 cells at days 56 and 70, respectively. F: Metastasis-free survival of mice following tail vein injection of either shCRTL or shGDF15#2 OMM1 cells. G: H&E staining of livers following tail vein injection of shCRTL or GDF15 silenced (GDF15 shRNA#2) OMM1 UM cells. Livers were collected after 7 weeks (shCRTL) or 11 weeks (shGDF15). Scale Bar = 6 mm. H: Quantification of liver metastases following tail vein injection of GDF15-expressing or silenced OMM1 cells (GDF15 shRNA#1 and shRNA#2).

Article Snippet: The effectiveness of blocking GDF15 on tube formation was assessed using an anti-GDF15 blocking antibody (ponsegromab: 120ng/ml, #HY-P99241, MedChemExpress, NJ, USA).

Techniques: Imaging, Injection, Staining, shRNA, Expressing

Journal: Cancer research

Article Title: GDF15 Reprograms the Microenvironment to Drive Liver Metastasis of Uveal Melanoma

doi: 10.1158/0008-5472.CAN-25-0536

Figure Lengend Snippet: A: H&E staining of livers from the eye to liver metastasis model, demonstrating fewer and smaller liver lesions following GDF15 shRNA silencing (shGDF15#1). B: Quantification of the size and number of liver metastases from A. C: IHC staining of liver metastases for fibronectin in MP41 tumors either expressing or with GDF15 silenced. D: Quantification of data from C. E: IHC staining of liver metastases from MP41 tumors for the ECM protein Collagen 1A1. F: Quantification of data from F. G: IHC staining of liver metastases for Ki67 in MP41 tumors either expressing or with GDF15 silenced. H: Quantification of data from G. I: IHC staining of liver metastases for the endothelial cell marker CD31 in MP41 tumors either expressing or with GDF15 silenced. J: Quantification of data from I.

Article Snippet: The effectiveness of blocking GDF15 on tube formation was assessed using an anti-GDF15 blocking antibody (ponsegromab: 120ng/ml, #HY-P99241, MedChemExpress, NJ, USA).

Techniques: Staining, shRNA, Immunohistochemistry, Expressing, Marker