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Updated: Jun 12, 2026
Macrophage Differentiation and Polarization into an M2-Like Phenotype using a Human Monocyte-Like THP-1 Leukemia Cell Line
Published on: August 2, 2021
Tumor-derived HMGB2 induces M2-like macrophage polarization via TRIM65-mediated NLRP3 degradation to promote DLBCL
Sanxiu He1, Yi Liu1, Yifeng Tang1
1Department of Hematology Oncology, Hematologic Oncology Intelligent Diagnosis and Treatment Engineering Research Center of Chongqing Education Commission of China, Chongqing Key Laboratory for the Mechanism and Intervention of Cancer Metastasis, Chongqing University Cancer Hospital, Chongqing, China.
Background:
Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin's lymphoma. Although standard immunochemotherapy is available, 30%-40% of patients develop refractory or relapsed disease, underscoring the need to elucidate the underlying mechanisms of therapy resistance. High mobility group box 2 (HMGB2) is overexpressed in DLBCL; however, its role in the pathogenesis of DLBCL and in mediating therapy resistance remains unclear.
Methods:
HMGB2 expression and prognostic value in DLBCL were analyzed using public databases, validated by quantitative reverse transcription-PCR and western blot analysis. In vivo and in vitro HMGB2 knockdown models were constructed to explore its role in DLBCL. Macrophage-DLBCL co-culture systems were established to investigate HMGB2-mediated macrophage polarization. Transcriptome sequencing was performed to identify downstream targets of HMGB2, with molecular mechanisms confirmed by western blot analysis, co-immunoprecipitation, and immunofluorescence. Exosome isolation and protease protection assays were used to determine the HMGB2 secretion pathway.
Results:
HMGB2 is overexpressed in DLBCL and correlates with poor prognosis. While HMGB2 knockdown showed no cell-intrinsic effects in vitro, it significantly suppressed tumor progression in vivo by remodeling the tumor microenvironment. We identified a novel mechanism whereby exosomal HMGB2 derived from DLBCL cells upregulates the E3 ubiquitin ligase tripartite motif containing 65 (TRIM65) in macrophages, promoting ubiquitin-mediated degradation of the NOD-like receptor protein 3 (NLRP3) inflammasome. This cascade drives M2-like macrophage polarization and impairs phagocytic function. Therapeutic targeting of this axis restored macrophage phagocytosis and synergized with CD20 immunotherapy to enhance its antitumor efficacy.
Conclusion:
Our study defines the HMGB2-TRIM65-NLRP3 axis as a pivotal immunosuppressive pathway in DLBCL. Targeting this axis represents a promising combinatory strategy to reprogram the tumor microenvironment and overcome therapy resistance.
