Forsøksdyr: Role of succinate in acute myeloid leukemia (Transfer to Tromsø)


Godkjenningsdato 08.09.2020

Godkjenningsperiode 08.09.2020-28.02.2023

The aim of this project is to study the contribution of succinate in acute myeloid leukemia (AML) development and to investigate possible treatments. Results will lead to advances in AML knowledge and potential patient treatment improvement. To explore the role of succinate in various aspects of hematopoetic stem cell (HSC) biology, we will assess the impact of succinate injections in the hematopoietic system of adult healthy and leukemic mice. Mice models knockout for succinate receptor (McCreath et al., 2015, Diabetes) are needed to examine whether Sucnr1 aids AML development and whether it could be a new therapeutic target. 5-fluorouracil (5-FU) challenge in C57BL/6 and Sucnr1 -/- mice will allow us to study whether succinate induces changes in HSC proliferation and self-renewal under stress hematopoiesis (Cheng T et al., 2000, Science). Despite mild toxicity of 5FU (Sanchez-Aguilera et al., 2014, Cell Stem Cell), this is an approved cancer treatment drug regularly used in humans. Our findings will reveal for the first time the contribution of SUCNR1 receptor in bone marrow (BM) microenvironment. Mice will be taken when they show early signs of frailty to prevent unnecessary suffering and a Kaplan-Meier curve will describe survival characteristics. C57BL/6, Sucnr1 -/- and Sucnr1 tomato mice will be used as recipient of control and mutant BM cells from Mx1-Cre NRAS-G12D mice. Recipient mice with a specific deletion of the Sucnr1 gene in the BM microenvironment will also be used as recipients of transplantation to understand the specific role of this receptor in the BM microenvironment. Conversely, succinate receptor deletion will be performed selectively in the hematopoietic system to understand the cell-autonomous effect in leukemia. Prior to transplantation, recipient mice need to be myeloablated through whole body irradiation. Total whole body irradiation is used in the field of blood stem cell function and cancer (for more information: http://www.bu.edu/orccommittees/iacuc/policies-and-guidelines/irradiation-of-rodents/). It allows to kill proliferating blood cells without significant damage of resting tissues, and simultaneously promotes proliferation of transplanted cells. Mice might suffer from a temporary lose of their body weight up to 26. Animals should be on their way to recovery at day 21 (cut-off weight loss 15%), and recovered at day 30 (cut-off weight loss 10%). All mice will be transplanted after irradiation, done in two half doses to reduce adverse effects. The results will provide a novel platform for more efficient therapies against AML, one of the most aggressive and frequent blood disorder that affects adults and children. It is not the goal of this study to analyze terminal stages.The number of animals will be kept in the minimum necessary (n=1020 total) to obtain statistically meaningful results, using them in the most efficient way possible.