The role of AKT inhibitors combined with Ruxolitinib in ameliorating myeloproliferative disorders in mice with CALR gene mutations
10.3760/cma.j.cn121090-20240920-00360
- VernacularTitle:AKT抑制剂联合芦可替尼对CALR基因突变骨髓增殖性肿瘤小鼠的治疗作用
- Author:
Liwei ZHANG
1
;
Qigang ZHANG
1
;
Mengchu JI
1
;
Kunming QI
1
;
Zhenyu LI
1
;
Kailin XU
1
;
Chunling FU
1
Author Information
1. 徐州医科大学血液病研究所，徐州医科大学附属医院血液科，徐州 221000
- Publication Type:Journal Article
- Keywords:
Myeloproliferative tumor;
Ruxolitinib;
AKT inhibitor;
CALR
- From:
Chinese Journal of Hematology
2025;46(8):750-757
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the combined therapeutic role of the AKT inhibitor MK2206 and Ruxolitinib in treating Myeloproliferative Neoplasms (MPN) driven by a calreticulin (CALR) gene mutation.Methods:① Murine bone marrow c-kit + cells were isolated by sacrificing mice and harvesting bone marrow from the femur, tibia, and ilium for subsequent c-kit + cell sorting. ② A CALR transplantation mouse model was established. GFP-tagged retroviral vectors containing either the CALR gene mutation or the migR1 control were constructed, packaged in Platinum-E cells, and used to transduce murine bone marrow c-kit + cells. These transduced cells were then transplanted into lethally irradiated female recipient mice via tail vein injection. ③ Following successful engraftment, the mice were randomly assigned to four treatment groups for intragastric administration. Complete blood counts were monitored periodically, and the spleen size and weight of transplanted mice were measured. ④ Flow cytometry was used to quantify the proportions of GFP + tumor cells, megakaryocytic lineage cells, and hematopoietic stem cells in both splenic and bone marrow tissues. Histopathological examination was performed to evaluate the degree of tumor cell infiltration in these organs. Results:① Following gavage treatment, peripheral blood platelet (PLT) and white blood cell counts were significantly lower in the combined AKT inhibitor MK2206 and Ruxolitinib group compared to the MK2206, Ruxolitinib, and control groups ( P<0.05). ② In comparison with the MK2206 and Ruxolitinib monotherapy groups, the combination therapy group exhibited a significant reduction in spleen weight and a marked improvement in splenomegaly at 30 weeks post-transplantation ( P<0.05). ③ After four weeks of continuous treatment, combined administration resulted in a significant decrease in the proportion of megakaryocytic lineage cells and GFP + tumor cells in the bone marrow and spleen ( P<0.05). Additionally, the proportion of hematopoietic stem cells in the bone marrow was also significantly reduced ( P<0.05). ④ Histopathological analysis (H&E staining) of bone marrow and spleen tissues confirmed that the combined regimen decreased both tumor cell infiltration and the proportion of abnormal megakaryocytes in these organs. Conclusion:The combination of AKT inhibitor MK2206 and Ruxolitinib is effective at significantly ameliorating disease symptoms and reducing tumor infiltration in vivo in mice with a myeloproliferative tumor transplantation driven by a CALR gene mutation.
