The CCL17-CCR4 axis is critical for mutant STAT6-mediated microenvironmental remodelling and therapeutic resistance in Relapsed/Refractory Diffuse Large B Cell Lymphoma

Abstract

Relapsed and refractory Diffuse Large B Cell Lymphoma (rrDLBCL) presents a significant challenge in hematology-oncology, with approximately 30-40% of DLBCL patients experiencing relapse or resistance to treatment. This underscores the urgent need to better understand the molecular mechanisms governing therapeutic resistance. Signal Transducer and Activator of Transcription 6 (STAT6) has been previously identified as a gene with recurrent D419 gain-of-function mutations in rrDLCBL. When STAT6D419 mutations are present in DLBCL tumour cells, we have demonstrated that transcription of the chemokine CCL17 (aka TARC) is increased, and tumours have increased infiltration of CD4+ T cells. However, the significance of increased T cell infiltration had not been determined. In the present study, we developed a mouse model of STAT6D419N mutant DLBCL, that recapitulates the critical features of human STAT6D419 mutant DLBCL, including increased expression of phospho-STAT6, increased CD4+ T cell invasion, and resistance to doxorubicin treatment. With this model, we found CD4+ T cells in STAT6D419N tumours have higher expression of the receptor for CCL17, CCR4. Using ex vivo functional assays we demonstrate that STAT6D419N tumour cells are directly chemoattractive to CCR4+ CD4+ T cells, and when CCR4 is inhibited using a small molecule antagonist, CD4+ T cells in STAT6D419N tumours are reduced and STAT6D419N tumours regain therapeutic sensitivity to doxorubicin. Using PhenoCycler imaging of human rrDLBCL samples, we find that STAT6D419 tumours indeed have increased expression of phospho-STAT6+ and increased cellular interactions between phospho-STAT6+ tumour cells and CD4+/ CCR4+ CD4+ T cells. Thus, our data identify CCR4 as an attractive therapeutic target in STAT6D419 mutant rrDLBCL.