Syngeneic tumor models in mice can be applied to evaluate antibodies, small molecules and vaccines. Triskelion offers syngeneic tumor models for melanoma (B16F10 in female C57BL/6 mice), colorectal cancer (CT26 in female Balb/c mice) and prostate cancer (TrampC2 in male C57BL/6 mice). Our syngeneic mouse models demonstrate sensitivity to treatment with well-known ICI (i.e. anti-PD-1, anti-CTLA-4). In addition, we develop tailor-made in vivo models with other tumor cell lines, which can be validated upon request with ICI (e.g. anti-PD1, anti-CTLA4 and anti-PDL1) available in house.
Readout parameters include but are not limited to:
- Tumor volume
- Advanced flow cytometry analysis of tumor infiltrating lymphocytes*
- Advanced flow cytometry analysis of lymphocyte subsets and myeloid cells in spleen, lymph nodes and blood*
- ELISPOT analysis of IFN-γ producing T cells
- Tumor histopathology (incl. semi-quantitative analysis of immune cell subsets).
- Immunohistochemistry of immune cells in the tumor (e.g.CD3)
- Body weight
* Our standard panels for flow cytometry analysis comprise the following markers:
CD3, CD8, CD44, CD62L, CD127, PD-1, Tim3, CD4, CD25, Foxp3, Tbet, Eomes, IFN-y, IL-2, TNF-α, CD11b, GR1, CD11c, F4/80 and CD19.
Human in vitro test system for immune checkpoint inhibitors
Triskelion offers a highly translatable human in vitro system to evaluate the efficacy and/or the mode of action of your drug candidates. Peripheral blood mononuclear cells (PBMC) from healthy donors are pre-incubated with your (combination of) ICI drug candidates. Subsequently, the PBMC are stimulated with superantigen (e.g. Staphyloccocus Enterotoxin B or anti-CD3/CD28). As a primary readout for (CD8+) T-cell activation, we measure cytokines, such as IL-2, in the culture supernatants with a Luminex-based method. ICI can amplify the production of IL-2 in this human translatable assay and thus Triskelion applies equivalents of Abatacept (CTLA4-Ig), Ipilimumab (anti-CTLA4) and Pembrolizumab (anti-PD1) as reference compounds.
Xenograft tumor models
Triskelion is at the forefront of developing tumor models which are translatable to the situation in cancer patients. We offer a humanized mouse model for skin cancer in which human MV-3 tumor cells are grafted into immune deficient NOG (NOD/Shi-scid/IL-2Rγnull) mice. Tumor-bearing mice receive peripheral blood mononuclear cells (PBMC) intravenously. Subsequently, the mice are subjected to anti-tumor therapy to evaluate the efficacy of your drug candidates.
These xenograft models represent a unique tool to evaluate the efficacy and the mode of action of drug candidates specifically targeting the human immune system and/or tumor tissue. Upon request, tailor-made humanized tumor models can be developed. We have ample experience with the analysis of the human T cells in the organs of xenografted mice, using flow cytometry and ElISPOT.
- Tumor volume
- Flow cytometry analysis of graft acceptance (PBMC)
- Advanced flow cytometry analysis of human lymphocytes in tumor and lymph nodes
- Tumor histopathology of the tumor (incl. semi-quantitative analysis of immune cells)
- Immunohistochemistry of immune cells in the tumor (e.g. human CD3)
- Body weight
Humanized tumor models with a complete human immune system
Triskelion has commenced the development of unique humanized tumor models for efficacy testing of cancer immunotherapies. Cancer patient tissues will be transplanted into immune-deficient recipient mice, together with human immune cells from the same donor (autologous grafting). These models will enable optimal translation of preclinical efficacy to the patient, which is lacking in the models currently available. Because these novel models will provide (key mechanistic) insight into human anti-tumor responses, they will accelerate the development of new drugs for cancer treatment.