Low dose ruxolitinib plus cyclosporine to treat murine immune-mediated bone marrow failure
Background and methods: Ruxolitinib (RUX), a Jak1/2 inhibitor, has been reported to attenuate murine bone marrow failure recently. Its potential toxocicty of anemia and thrombocytopenia in human remains a concern. To minimize its potential toxoxicity, we tested therapeutic effectsof low dose ruxolitinib plus cyclosporine in murine model of immune-mediated bone marrow failure. Bone marrow CD8 and CD4 T cells were sorted from treated or untreated bone marrow failure mice. RNA-Seq and analysis was performed using SMART-Seq mRNA LP Kit (Takara) and the Illumina Novaseq6000, according to the Institute's protocols. Results: low dose of ruxolitinib plus cyclosporine improved pancytopenia and BM cellularity and decreased BM T cell infiltration in bone marrow failure mice. RNA sequencing demonstrated that low dose of ruxolitinib plus cyclosporine suppressed immune-related pathways in bone marrow infiltrated CD8 T cells and MHC-II expression in CD4 T cells compared with untreated mice. Conclusion: Our results demonstrate that low dose of ruxolitinib plus cyclosporine remains the efficacy in attenuation of disease and extending survival of immune-mediated bone marrow failure mice. Overall design: We compared transcriptomes of CD8 and CD4 T cells in the bone marrow failure mice treated with and without low dose of ruxolitinib plus cyclosporine, and described differentially expressed genes of these cells. CD8+ and CD4+ T cells were isolated from bone marrow cells of low dose of ruxolitinib plus cyclosporine-treated and control BMF mice by FACS-sorting. To get enough cells for RNA extraction, CD8+ and CD4+ T cells from every 2-3 mice in the same group were pooled together, we obtained 3 pools/group.