Efficacy And Molecular Mechanism Of PIKfyve And Tubulin Dual-target Inhibitors HZX-02-059-01 On Double-Hit Lymphoma

Double-hit lymphoma(DHL)is a specific subtype of diffuse large B-cell lymphoma(DLBCL),which is characterized by chromosome rearrangement of MYC,bcl-2,or bcl-6.DHL patients are not sensitive to the current first-line R-CHOP chemotherapy and generally have poor prognosis.The intense chemotherapy improves the progression-free survival of DHL to some extent,but the prognosis of DHL is still worse than that of other DLBCL.In addition,intense chemotherapy can also bring more serious side effects,increasing the treatment-related mortality of patients.Therefore,finding effective and safe treatment strategies is an urgent problem to be solved in the treatment of DHL patients.At present,PIKfyve inhibitors have a significant killing effect on non-hodgkin’s lymphoma(NHL),but no further progress has been made.In this study,a novel small molecule compound HZX-02-059-01 was taken as the research object.Its structure is different from all the reported PIKfyve inhibitors,and it has two prominent functional targets,PIKfyve and tubulin.In order to explore its effect on killing DHL and explore its mechanism,it is expected to provide new ideas and directions for the treatment of DHL.In this study,WILL-2,LR and TMD8 cell lines were used as research objects to investigate the killing effect and molecular mechanism of dual-target inhibitory compound HZX-02-059-01 on DHL.The significant proliferation inhibition effect of HZX-02-059-01 on DHL was verified by cell-counting kit 8.The IC50 of WILL-2,LR and TMD8 cells were 106.9 nMol/L,186 nMol/L and 257.7 nMol/L,respectively,after 72H treatment.It was further demonstrated that HZX-02-059-01 could induce caspase-independent cell death,instead induce the methuosis of DHL cells,which was confirmed by the down-regulation of the PIKfyve/mTOR/TFEB pathway detected by Western Blotting.Flow cytometry showed that HZX-02-059-01 could block DHL cells in G2/M phase,while Western Blotting suggested decreased C-MYC expression.In vivo experiments further confirmed the killing effect of HZX-02-059-01 on DHL.In addition,HZX-02-059-01 can also reduce the expression of PD-L1 and CD47 to avoid immune escape.The results of this study indicate that HZX-02-059-01 has a significant killing effect on DHL in vitro and in vivo,which can induce the methuosis of DHL cells and affect their cell cycle distribution.In summary,the present findings suggest that HZX-02-059-01 might represent a potential candidate agent for DHL treatment.