Pre-clinical evaluation of CAR Treatment in solid tumor
Purpose/expected benefit
This project identifies candidates for the use of chimeric antigen receptor (CAR) T cells in adoptive T cell therapy in the treatment of cancer patients. Adoptive cell therapy involves the administration of a large number of selected cells with high avidity and specificity for tumour antigen. The chimeric antigen receptor (CAR) has been identified, cloned and tested in vitro. We have shown that CAR T cells efficiently kill tumour cell lines corresponding to the tumour type the CAR is directed against. The potential for CAR gene transfer must be demonstrated in vivo before it can be transferred to the clinic and is a requirement from SLV. Efficacy in tumour eradication and other parameters will be monitored and will be essential information for the design of future clinical trials.
Distress
The project is classified to inflict moderate load of distress on the animals. The treatment will not cause the animals much discomfort. Both tumour and treatment with redirected T cells will be administered using syringe, any discomfort last for seconds. Tumour growth is monitored weekly with IVIS performed or MR under gas anaesthesia. Tumour development may impair the general health status to the animals, the animals' general health status is monitored daily and the animals will be killed if found to have a large tumour load or impaired health according to given criteria.
Number of animals
In total we will need 740 NOD SCID mice.
3R
Replacement; Testing the potential of the selected CAR in vivo is a requirement before these protocols can be transferred to the clinic. Reduction; we use a minimal number of animals that will support a statistically significant result. We use a tumour model that has been modified with a luciferase reporter system , which allows us to monitor tumour development in a gentle way using the in-vivo imaging system (IVIS), and thus a reduction in the number of animals in each group. In certain models we also use non-modified patient-derived xenografts (PDX) that are slow growing and injected intraperitoneally. In the PDX models, tumour growth is monitored using MR as the tumours do not contain the luciferase gene. Refinement; the animal's cage is equipped with environmental enrichment such as paper for nest construction (always) and alternation between cardboard houses, plastic pipes or plastic igloo.
This project identifies candidates for the use of chimeric antigen receptor (CAR) T cells in adoptive T cell therapy in the treatment of cancer patients. Adoptive cell therapy involves the administration of a large number of selected cells with high avidity and specificity for tumour antigen. The chimeric antigen receptor (CAR) has been identified, cloned and tested in vitro. We have shown that CAR T cells efficiently kill tumour cell lines corresponding to the tumour type the CAR is directed against. The potential for CAR gene transfer must be demonstrated in vivo before it can be transferred to the clinic and is a requirement from SLV. Efficacy in tumour eradication and other parameters will be monitored and will be essential information for the design of future clinical trials.
Distress
The project is classified to inflict moderate load of distress on the animals. The treatment will not cause the animals much discomfort. Both tumour and treatment with redirected T cells will be administered using syringe, any discomfort last for seconds. Tumour growth is monitored weekly with IVIS performed or MR under gas anaesthesia. Tumour development may impair the general health status to the animals, the animals' general health status is monitored daily and the animals will be killed if found to have a large tumour load or impaired health according to given criteria.
Number of animals
In total we will need 740 NOD SCID mice.
3R
Replacement; Testing the potential of the selected CAR in vivo is a requirement before these protocols can be transferred to the clinic. Reduction; we use a minimal number of animals that will support a statistically significant result. We use a tumour model that has been modified with a luciferase reporter system , which allows us to monitor tumour development in a gentle way using the in-vivo imaging system (IVIS), and thus a reduction in the number of animals in each group. In certain models we also use non-modified patient-derived xenografts (PDX) that are slow growing and injected intraperitoneally. In the PDX models, tumour growth is monitored using MR as the tumours do not contain the luciferase gene. Refinement; the animal's cage is equipped with environmental enrichment such as paper for nest construction (always) and alternation between cardboard houses, plastic pipes or plastic igloo.