Evaluation of in situ expression and anti-tumor efficacy of plasmid DNA encoded immunotherapy against tumors in Balb/c nu/nu mice
1 Purpose
Nykode Therapeutics AS is dedicated to the discovery and development of novel cancer immunotherapies. Nykode’s technology platform is highly adaptable and can be modified for expression of a wide range of therapeutic molecules. The most impactful immunotherapies against cancer targets the immune checkpoint molecules CTLA-4 and PD-1/PD-L1(1,2) or mutated receptors expressed on cancer cells, and this large field of therapy now encompasses several drugs approved by the FDA for their ability to diminish tumor growth by inducing immune responses against tumor(s)(3–5). The purpose of this experiment is to evaluate new plasmid DNA-based immunotherapy in terms of secretion efficiency and anti-tumor efficacy
2 Distress
The pDNA will be delivered by i.m. injection followed by electroporation using a short pulse pattern which gives less pain and soreness after awakening. To increase transfection efficiency and protein production, we will make use of the adjuvant Hyaluronidase. We expect low systemic levels of product and low toxicity as similar therapies have been tested in mice extensively (4,6). The animals will experience short-term stress/discomfort during blood sampling. The mice will be injected with a lethal dose of tumor cells, and the experiment is classified as moderate in severity. We have long experience with subcutaneous tumor models and expect subcutaneous tumor growth to cause only mild-to-moderate stress to the animals.
3 Expected benefit
Immunotherapy have proven to have an unprecedented impact in the field of cancer immunotherapy as well the treatment of autoimmune- and infectious diseases. We will employ Nykode’s technology platform for delivery of immunotherapy in situ. The proof-of-principle data that we will generate through pre-clinical studies in mouse models will provide mechanistic insight into the secretion level of our pDNA constructs and their potential anti-tumor effect. The pre-clinical experiments will provide an opportunity for Nykode to submit applications to conduct DNA immunotherapy in clinical trials and provide a new line of treatment opportunities for patients.
4 Number of animals, and what kind
In this experiment we will use 410 BALB/c nu/nu mice (Mus musculus).
5 How to adhere to 3R
Animal experiments are the only available option to evaluate secretion, systemic circulation and the anti-tumor efficacy of the DNA constructs against tumors. In vitro experiments will be performed to evaluate a panel of constructs for their secretion levels and capacity to bind mutated/overexpressed target receptor. To further reduce the number of animals used, we include as few animals per group as is necessary to observe and interpret differences between the treatment groups. The animals will be sedated during the procedures and only mild short-term pain is expected, and the animals will be monitored closely over the entire experiment.
Nykode Therapeutics AS is dedicated to the discovery and development of novel cancer immunotherapies. Nykode’s technology platform is highly adaptable and can be modified for expression of a wide range of therapeutic molecules. The most impactful immunotherapies against cancer targets the immune checkpoint molecules CTLA-4 and PD-1/PD-L1(1,2) or mutated receptors expressed on cancer cells, and this large field of therapy now encompasses several drugs approved by the FDA for their ability to diminish tumor growth by inducing immune responses against tumor(s)(3–5). The purpose of this experiment is to evaluate new plasmid DNA-based immunotherapy in terms of secretion efficiency and anti-tumor efficacy
2 Distress
The pDNA will be delivered by i.m. injection followed by electroporation using a short pulse pattern which gives less pain and soreness after awakening. To increase transfection efficiency and protein production, we will make use of the adjuvant Hyaluronidase. We expect low systemic levels of product and low toxicity as similar therapies have been tested in mice extensively (4,6). The animals will experience short-term stress/discomfort during blood sampling. The mice will be injected with a lethal dose of tumor cells, and the experiment is classified as moderate in severity. We have long experience with subcutaneous tumor models and expect subcutaneous tumor growth to cause only mild-to-moderate stress to the animals.
3 Expected benefit
Immunotherapy have proven to have an unprecedented impact in the field of cancer immunotherapy as well the treatment of autoimmune- and infectious diseases. We will employ Nykode’s technology platform for delivery of immunotherapy in situ. The proof-of-principle data that we will generate through pre-clinical studies in mouse models will provide mechanistic insight into the secretion level of our pDNA constructs and their potential anti-tumor effect. The pre-clinical experiments will provide an opportunity for Nykode to submit applications to conduct DNA immunotherapy in clinical trials and provide a new line of treatment opportunities for patients.
4 Number of animals, and what kind
In this experiment we will use 410 BALB/c nu/nu mice (Mus musculus).
5 How to adhere to 3R
Animal experiments are the only available option to evaluate secretion, systemic circulation and the anti-tumor efficacy of the DNA constructs against tumors. In vitro experiments will be performed to evaluate a panel of constructs for their secretion levels and capacity to bind mutated/overexpressed target receptor. To further reduce the number of animals used, we include as few animals per group as is necessary to observe and interpret differences between the treatment groups. The animals will be sedated during the procedures and only mild short-term pain is expected, and the animals will be monitored closely over the entire experiment.