Antitumor activity of FGF conjugated with Auristatin and Amanitin
1 Purpose
FGFR (Fibroblast Growth Factor Receptor) is overexpressed in many cancer types and is associated with the metastatic process in lung and breast cancer. Simultaneously, FGFR has a very low expression in healthy tissue, making it a very good target for cancer therapy. In this project we will test the antitumor activity of FGF2 (which binds specifically to FGFR) conjugated with two drugs simultaneously: auristatin and amanitin.
2 Distress
The experiments performed in this application involve procedures where the animals are given injections of tumor cells, followed by IV injections of the FGF2-drug conjugates. The dosage of the compounds will be adjusted to an acceptable level with minimal and tolerable side effects.
3 Expected benefit
Many cancers, including breast and lung cancer, have high levels of FGFR. Our approach can potentially provide a new and highly directed therapy, with little to no effect on healthy tissue, against these cancer types. This falls within the aims of personalized medicine and moves away from the traditional chemotherapy approach with generalized side effects for the patient. We will also use breast and lung cancer xenografts with varying levels of FGFR, as well as patient derived xenografts (PDXs). Using PDXs will allow us to obtain data more readily translated into human medicine.
4 Number of animals, and what kind
126 nude mice
70 NXG mice
5 How to adhere to 3R
Replacement:
We will test several variants of FGF2-drug conjugates in cell lines in vitro in order to select the one that is most potent against cancer cells and simultaneously the least harmful for healthy tissue. Only the most promising candidate will be used in the animal experiments.
Reduction:
Based on previous experience, the different groups have the minimum number of animals to achieve significant results. Also, we reduced the test groups from seven to two (control and treatment group). We will first use a reduced group of animals to identify the dosage that is expected to have an antitumor effect with as little as possible adverse effects on the health of the animals. We will also base our dosage levels on previously published research that used a similar FGF2-auristatin conjugate (FOTS Id 13177).
Refinement:
Anesthetic and analgesic regimes for pain relief are in place to reduce potential pain after implantation of tumor tissue. Appropriate experimental endpoints that allow for early intervention have been implemented to reduce animal suffering. Handling of the animals will be performed by experienced personnel so that the discomfort/stress to the mice will be at a minimum. The environment of the mice will be enriched with cardboard houses, plastic tubes, and/or shredded paper and allow for natural hiding, burrowing and nesting behavior..
FGFR (Fibroblast Growth Factor Receptor) is overexpressed in many cancer types and is associated with the metastatic process in lung and breast cancer. Simultaneously, FGFR has a very low expression in healthy tissue, making it a very good target for cancer therapy. In this project we will test the antitumor activity of FGF2 (which binds specifically to FGFR) conjugated with two drugs simultaneously: auristatin and amanitin.
2 Distress
The experiments performed in this application involve procedures where the animals are given injections of tumor cells, followed by IV injections of the FGF2-drug conjugates. The dosage of the compounds will be adjusted to an acceptable level with minimal and tolerable side effects.
3 Expected benefit
Many cancers, including breast and lung cancer, have high levels of FGFR. Our approach can potentially provide a new and highly directed therapy, with little to no effect on healthy tissue, against these cancer types. This falls within the aims of personalized medicine and moves away from the traditional chemotherapy approach with generalized side effects for the patient. We will also use breast and lung cancer xenografts with varying levels of FGFR, as well as patient derived xenografts (PDXs). Using PDXs will allow us to obtain data more readily translated into human medicine.
4 Number of animals, and what kind
126 nude mice
70 NXG mice
5 How to adhere to 3R
Replacement:
We will test several variants of FGF2-drug conjugates in cell lines in vitro in order to select the one that is most potent against cancer cells and simultaneously the least harmful for healthy tissue. Only the most promising candidate will be used in the animal experiments.
Reduction:
Based on previous experience, the different groups have the minimum number of animals to achieve significant results. Also, we reduced the test groups from seven to two (control and treatment group). We will first use a reduced group of animals to identify the dosage that is expected to have an antitumor effect with as little as possible adverse effects on the health of the animals. We will also base our dosage levels on previously published research that used a similar FGF2-auristatin conjugate (FOTS Id 13177).
Refinement:
Anesthetic and analgesic regimes for pain relief are in place to reduce potential pain after implantation of tumor tissue. Appropriate experimental endpoints that allow for early intervention have been implemented to reduce animal suffering. Handling of the animals will be performed by experienced personnel so that the discomfort/stress to the mice will be at a minimum. The environment of the mice will be enriched with cardboard houses, plastic tubes, and/or shredded paper and allow for natural hiding, burrowing and nesting behavior..