富FAM76B组氨酸区缺失通过液-液相分离增强巨噬细胞介导的骨肉瘤抑制

IF 4.7 2区医学 Q2 IMMUNOLOGY

International immunopharmacology Pub Date : 2025-08-05 DOI:10.1016/j.intimp.2025.115289

Tiansheng Zhou , Xiangyu Yang , Ding Zhao , Xiaoqian Tan , Yingqi Liu , Tian Li , Guanghui Zhu

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引用次数: 0

摘要

骨肉瘤（OS）是一种主要发生于年轻人的常见恶性肿瘤，由于骨髓细胞介导的免疫抑制微环境，它经常逃避基于T细胞的免疫治疗。本研究探讨FAM76B在OS中的作用及其对巨噬细胞极化和肿瘤免疫应答的影响。方法利用荧光共聚焦显微镜和光漂白后荧光恢复（FRAP）实验，预测FAM76B的内在无序区（IDRs），并探讨其在体外的相分离性能。同时进行免疫荧光染色。我们利用CRISPR Cas9和位点定向诱变技术改造骨肉瘤细胞，研究FAM76B相分离对细胞功能的影响。FAM76B的作用通过菌落形成、MTT试验、伤口愈合试验、巨噬细胞功能和极化的流式细胞术、T细胞和NK细胞活性、皮下肿瘤异种移植模型和免疫组织化学来评估。结果FAM76B重编程巨噬细胞的缺失，增强了OS患者的抗肿瘤免疫功能。FAM76B缺失逆转免疫抑制，导致T细胞浸润和活化增加。结果发现FAM76B具有组氨酸富区（HRR）相分离特性；该区域的缺失增强了其诱导M1样巨噬细胞的能力，抑制了OS细胞的生长。结论针对FAM76B相分离可能提供一种新的治疗方法来对抗肿瘤诱导的免疫抑制，提高T细胞免疫治疗骨肉瘤和其他实体瘤的疗效。

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Deletion of FAM76B histidine-rich region enhances macrophage-mediated osteosarcoma inhibition via liquid-liquid phase separation

Background

Osteosarcoma (OS), a prevalent malignancy primarily in young individuals, often evades T cell based immunotherapy due to an immunosuppressive myeloid cell-mediated microenvironment. This study investigated the role of FAM76B in OS and its impact on macrophage polarization and tumor immune response.

Methods

We predicted intrinsically disordered regions (IDRs) of FAM76B and explored its phase separation properties in vitro using fluorescence confocal microscopy and fluorescence recovery after photobleaching (FRAP) experiments. Immunofluorescence staining was also performed. We engineered osteosarcoma cells using CRISPR Cas9 and site directed mutagenesis to study the impact of FAM76B phase separation on cellular function. The effects of FAM76B were assessed through colony formation, MTT assays, wound healing assays, flow cytometry for macrophage function and polarization, T cell and NK cell activity, subcutaneous tumor xenograft models, and immunohistochemistry.

Results

Deletion of FAM76B reprogrammed macrophages, enhancing antitumor immunity in OS. Absence of FAM76B reversed immunosuppression, leading to increased T cell infiltration and activation. We found FAM76B exhibited histidine rich region (HRR) dependent phase separation; deletion of this region enhanced its ability to induce M1 like macrophages, inhibiting OS cell growth.

Conclusion

Our findings suggest that targeting FAM76B phase separation may offer a novel therapeutic approach to counteract tumor induced immunosuppression and enhance efficacy of T cell based immunotherapy in osteosarcoma and potentially other solid tumors.

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来源期刊

International immunopharmacology 医学-免疫学

CiteScore

8.40

自引率

3.60%

发文量

935

审稿时长

53 days

期刊介绍： International Immunopharmacology is the primary vehicle for the publication of original research papers pertinent to the overlapping areas of immunology, pharmacology, cytokine biology, immunotherapy, immunopathology and immunotoxicology. Review articles that encompass these subjects are also welcome. The subject material appropriate for submission includes: • Clinical studies employing immunotherapy of any type including the use of: bacterial and chemical agents; thymic hormones, interferon, lymphokines, etc., in transplantation and diseases such as cancer, immunodeficiency, chronic infection and allergic, inflammatory or autoimmune disorders. • Studies on the mechanisms of action of these agents for specific parameters of immune competence as well as the overall clinical state. • Pre-clinical animal studies and in vitro studies on mechanisms of action with immunopotentiators, immunomodulators, immunoadjuvants and other pharmacological agents active on cells participating in immune or allergic responses. • Pharmacological compounds, microbial products and toxicological agents that affect the lymphoid system, and their mechanisms of action. • Agents that activate genes or modify transcription and translation within the immune response. • Substances activated, generated, or released through immunologic or related pathways that are pharmacologically active. • Production, function and regulation of cytokines and their receptors. • Classical pharmacological studies on the effects of chemokines and bioactive factors released during immunological reactions.