• Origin: The MC38 cell line is derived from C57BL6 murine colon adenocarcinoma cells. The cell line is a commonly used murine model for colorectal carcinoma.
• Background Information: Growth differentiation factor 15 (GDF-15) is a cytokine linked to anorexia and weight loss in preclinical models and is associated with cachexia and poor survival in cancer patients. It regulates energy balance by activating the glial cell-derived neurotrophic factor receptor alpha-like (GFRAL), which is found in the area postrema (AP) and nucleus of the solitary tract (NTS) in the hindbrain.
• Gene targeting strategy: The exogenous promoter and human GDF15 coding sequence were inserted to replace part of murine exon 1 and all of exons 2. The insertion disrupts the endogenous murine Gdf15 gene, resulting in a non-functional transcript.
• Tumorigenicity: Confirmed in B-hGDF15 mice.
• Application: The B-hGDF15 MC38 tumor models can be used for preclinical evaluation of monoclonal antibody drugs and bispecific antibody drugs targeting human GDF15.
• Notes: 
• The B-hGDF15 MC38 tumor was only implanted in B-hGDF15 mice and has not been implanted in C57BL/6 mice.

GDF15 expression analysis in B-hGDF15 MC38 cells by ELISA. Cell culture supernatant was collected from wild-type MC38 and B-hGDF15 MC38 #2-B06. Expression level of mouse and human GDF15 were analyzed by ELISA (Mouse/Rat GDF-15 Quantikine ELISA Kit: R&D, MGD150; Human GDF-15 Quantikine ELISA Kit: R&D, DGD150). Human GDF15 was exclusively detectable in homozygous B-hGDF15 MC38. The 2-B06 clone of B-hGDF15 MC38 cells was used for in vivo experiments.

Subcutaneous tumor growth of B-hGDF15 MC38 cells. B-hGDF15 MC38 cells (5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into homozygous B-hGDF15 mice (female, 6-8 weeks old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume. (B) Body weight. The B-hGDF15 MC38 cell inoculation group shows a decreasing trend in body weight compared to the wild-type MC38 cell inoculation group. Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². Results indicate that B-hGDF15 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM. 

Note: The B-hGDF15 MC38 tumor was only implanted in B-hGDF15 mice and has not been implanted in C57BL/6 mice.

GDF15 expression evaluated on B-hGDF15 MC38 tumor cells by ELISA. B-hGDF15 MC38 cells were subcutaneously transplanted into homozygous B-hGDF15 mice (n=6). Upon conclusion of the experiment, serum was collected from homozygous B-hGDF15 mice. As shown, human GDF15 was highly expressed of tumor cells. Therefore, B-hGDF15 MC38 cells can be used for in vivo efficacy studies evaluating novel GDF15 therapeutics.

Antitumor activity of anti-human GDF15 antibody (ponsegromab analog, in-house) in B-hGDF15 mice. (A) B-hGDF15 MC38 tumor growth in B-hGDF15 mice. Murine colon cancer B-hGDF15 MC38 cells (5×10⁵) were subcutaneously implanted into homozygous B-hGDF15 mice (female, 9 weeks old, n=6). Mice were grouped when tumor volume reached approximately 100-150 mm³, at which time they were intraperitoneally injected with anti-human GDF15 antibody indicated in panel. (B) Body weight changes during treatment. (C) The weight change of each mouse in the group compared to Day 0. As shown in panel B and C, anti-human GDF15 antibody was effective in controlling the weight loss induced by B-hGDF15 MC38 in B-hGDF15 mice, demonstrating that the B-hGDF15 mice and B-hGDF15 MC38 provide a powerful preclinical model for in vivo evaluation of anti-human GDF15 antibodies. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test.  *P < 0.05, **P < 0.01, ***P < 0.001. 

The overage of this tumor model is 50%.