IRF4 contributes to chemoresistance in IGH::BCL2-positive diffuse large B-cell lymphomas by mediating BCL2-induced SOX9 expression

IF 7.9 1区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Clinical and Translational Medicine Pub Date : 2025-05-12 DOI:10.1002/ctm2.70336

Yirong Zhang, Zizhen Xu, Ruixin Sun, Yixuan Gao, Innocent Agida, Kasimujiang Aximujiang, Lin Yuan, Jiao Ma

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Abstract

Background

Diffuse large B-cell lymphoma (DLBCL), an aggressive type of non-Hodgkin's lymphoma, has a high relapse/refractory rate. We previously identified sex-determining region Y (SRY)-box transcription factor (SOX9) as a transcription factor that serves as a prognostic biomarker, particularly in BCL2-overexpressing DLBCL, and plays a vital role in lymphomagenesis. However, the molecular mechanisms that modulate the aberrant expression of SOX9 in this DLBCL subset remain unknown.

Methods

Cell viability, apoptosis and cell cycle assays were performed to determine whether SOX9 contributes to DLBCL chemoresistance and rescues silencing IRF4-induced phenotypes. Protein‒protein interactions and protein ubiquitination were elucidated using immunoprecipitation, immunohistochemistry, immunofluorescence and immunoblotting. Chromatin immunoprecipitation sequencing (ChIP-seq), ChIP and dual-luciferase reporter assays were used to investigate IRF4 binding to the SOX9 promoter. The therapeutic potential of IRF4 inhibition was evaluated in vitro and in a mouse model of DLBCL xenografts.

Results

SOX9 enhanced the resistance of the BCL2-overexpressing DLBCL subset to chemotherapy or a BCL2 inhibitor. Moreover, BCL2 inhibition downregulated SOX9 in an immunoglobulin heavy chain/BCL2-positive DLBCL subset. We further identified IRF4 as a key regulator of BCL2-induced SOX9 expression, and ChIP-seq confirmed that IRF4 is a key transcription factor for SOX9 in DLBCL. In addition, BCL2 promotes IRF4 entry into the nucleus by enhancing protein stability and downregulating proteasomal ubiquitination, thereby enforcing SOX9-mediated phenotypes. Finally, in a DLBCL cell line and xenografted mouse model, in vivo inhibition of IRF4 with an hIRF4 antisense oligonucleotide repressed lymphomagenesis and DLBCL chemoresistance.

Conclusions

Our data support the conclusion that IRF4 plays an essential role in BCL2-induced upregulation of SOX9 expression, and targeting IRF4 may represent a promising therapeutic strategy to cure relapsed and refractory DLBCL.

Keypoints/Highlights

BCL2 activated IRF4 by enhancing its nuclear activity to induce sex-determining region Y (SRY)-box 9 protein (SOX9) aberrant expression, which is a critical pathway for drug resistance in BCL2-overexpressing diffuse large B-cell lymphoma (DLBCL).
Targeting IRF4 may be worth investigating further regarding its potential to overcome the chemoresistance of BCL2-overexpressing DLBCL to standard therapies.

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IRF4通过介导bcl2诱导的SOX9表达参与IGH:: bcl2阳性弥漫大b细胞淋巴瘤的化疗耐药

弥漫性大b细胞淋巴瘤（DLBCL）是一种侵袭性非霍奇金淋巴瘤，具有很高的复发/难治率。我们之前发现性别决定区Y (SRY)-box转录因子（SOX9）是一种转录因子，可作为预后生物标志物，特别是在bcl2过表达的DLBCL中，并在淋巴瘤发生中起重要作用。然而，调控SOX9在DLBCL亚群中异常表达的分子机制尚不清楚。方法通过细胞活力、细胞凋亡和细胞周期测定来确定SOX9是否有助于DLBCL化疗耐药，并挽救沉默irf4诱导的表型。采用免疫沉淀、免疫组织化学、免疫荧光和免疫印迹等方法对蛋白相互作用和蛋白泛素化进行了分析。采用染色质免疫沉淀测序（ChIP-seq）、ChIP和双荧光素酶报告基因检测来研究IRF4与SOX9启动子的结合。在体外和DLBCL异种移植小鼠模型中评估了IRF4抑制的治疗潜力。结果SOX9增强了过表达BCL2的DLBCL亚群对化疗或BCL2抑制剂的耐药性。此外，BCL2抑制下调了免疫球蛋白重链/BCL2阳性DLBCL亚群中的SOX9。我们进一步发现IRF4是bcl2诱导SOX9表达的关键调节因子，ChIP-seq证实IRF4是DLBCL中SOX9的关键转录因子。此外，BCL2通过增强蛋白质稳定性和下调蛋白酶体泛素化来促进IRF4进入细胞核，从而加强sox9介导的表型。最后，在DLBCL细胞系和异种移植小鼠模型中，体内用hIRF4反义寡核苷酸抑制IRF4抑制淋巴瘤发生和DLBCL化疗耐药。我们的数据支持IRF4在bcl2诱导的SOX9表达上调中发挥重要作用的结论，靶向IRF4可能是治疗复发和难治性DLBCL的一种有希望的治疗策略。BCL2通过增强IRF4核活性激活IRF4，诱导性别决定区Y (SRY)-box 9蛋白（SOX9）异常表达，这是BCL2过表达弥漫大b细胞淋巴瘤（DLBCL）耐药的关键途径。靶向IRF4可能值得进一步研究其克服bcl2过表达的DLBCL对标准治疗的化疗耐药的潜力。

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来源期刊

Clinical and Translational Medicine Multiple-

CiteScore

15.90

自引率

1.90%

发文量

450

审稿时长

4 weeks

期刊介绍： Clinical and Translational Medicine (CTM) is an international, peer-reviewed, open-access journal dedicated to accelerating the translation of preclinical research into clinical applications and fostering communication between basic and clinical scientists. It highlights the clinical potential and application of various fields including biotechnologies, biomaterials, bioengineering, biomarkers, molecular medicine, omics science, bioinformatics, immunology, molecular imaging, drug discovery, regulation, and health policy. With a focus on the bench-to-bedside approach, CTM prioritizes studies and clinical observations that generate hypotheses relevant to patients and diseases, guiding investigations in cellular and molecular medicine. The journal encourages submissions from clinicians, researchers, policymakers, and industry professionals.