Forsøksdyr: implantation of mammary tumor cells in Foxp3-RFP+AKT3-/- transgenic mice mice


Godkjenningsdato 12.01.2018

1. Purpose of the study.
In this project we want first to assess the role of AKT3 in both tumor development and metastasis. We want to implant tumor cells in the mammary gland and tail vein (metastasis) in Foxp3-RFP+AKT3-/- transgenic mice (FOTS 9596) and compare it with wild type mice.
The second purpose is to use the AKT3 null mice to characterize and isolate immune cells lacking Akt3 and compare those cells with wild type immune cells expressing AKT3.
The third purpose of the study will be to characterize and isolate immune cells lacking Akt3 in the presence of tumor.
The last aim of the study will be to use AKT3 null mice to test check points inhibitor treatment and to see if the absence of AKT3 potentiate those treatments
2. Expected harm/severity for the animals.
A part of the project will consist in acute experiments were we will characterize immune cells lacking AKT3, mostly by flow cytometry in both normal and cancer conditions. We will implant tumor cells in mammary gland or tail vein, the animals will be monitored twice a week and the experiment will be stopped when distress is observed. The tumor implantation will be done under gas anesthesia.
Check point inhibitors treatments will consist in IP injections twice a week.
3. Expected benefit for science or society
Axl inhibitors are into clinical trials, we aim to prove that as downstream effector of axl, inhibition of AKT3 could be an additional therapeutic target. AKT3 will be as well used to test check points inhibitor treatment and to see if the absence of AKT3 potentiate those treatments. The checkpoints inhibitors are new promising treatments,we may potentiate those treatment and increase the responders by associating it with akt3 inhibitors.
We will use the AKT3 null mice to first characterize and isolate immune cells lacking Akt3 and compare those cells with wild type immune cells expressing AKT3 in physiological conditions or in the presence of tumor cells.
4. How many and what kind of animals will be used
We will need 360 animals, it will be mus musculus Foxp3-RFP+AKT3-/- transgenic mice (fots 9596) in a C57bl6/J, in addition wild type control mice will be ordered.
5. How are compliance with the requirements for replacement, reduction and refinement safe guarded.
The purpose of this experiment is to use a mouse strain, in which expression of Akt3 is suppressed (Fots 9596). Akt3 expression differs between individuals. To understand the importance of akt3 in developing animals and in tumor development it is necessary to perform in vivo studies, since in vitro studies cannot mimic the body complexity.
This project aim to assess different aspects of AKT3 biology. We will do the experiments sequentially, we will start by studying the role of akt3 in normal immune system, then we will implant tumors and asses the same immune cells with mammary tumor cells.
At the end of the project we will test checkpoints inhibitors in AKT3 null mice and compare it with wild type mice.