上海交通大学学报（医学版）2025，Vol.45Issue(9)：1126-1137,12.DOI:10.3969/j.issn.1674-8115.2025.09.005
PIKfyve抑制剂靶向治疗STING通路缺陷黑色素瘤的机制研究
Mechanistic study of targeting melanoma with STING pathway deficiencies via PIKfyve inhibitor
摘要
Abstract
Objective·To explore the antitumor effects and potential mechanisms of combining phosphoinositide 3-kinase,FYVE-type zinc finger containing(PIKfyve)inhibitor YM201636 with the stimulator of interferon genes(STING)agonist diABZI in STING pathway-deficient melanoma.Methods·The mRNA and protein expression levels of STING in human cancer cell lines were obtained from the Cancer Cell Line Encyclopedia(CCLE)and UniProt databases.Based on median expression values,melanoma cell lines with high STING mRNA but low protein expression were identified.Quantitative real-time PCR(qRT-PCR)and Western blotting were performed to validate STING mRNA and protein expression in human melanoma cells.The murine melanoma cell line YUMM1.7,characterized by low STING protein expression,was selected through Western blotting.The ability of YM201636 to restore STING protein expression in YUMM1.7 cells was evaluated.STING agonist diABZI was then applied in combination with YM201636 to analyze the synergistic tumor cell-killing effect through CCK-8 assay.Western blotting was used to detect the phosphorylation of TANK-binding kinase 1(TBK1)and interferon regulatory factor 3(IRF3),and qRT-PCR was used to evaluate type Ⅰ interferon expression.A mouse melanoma model was established and treated with YM201636,diABZI,or their combination.Tumor volume was measured,and treatment efficacy was assessed.RNA sequencing and immunofluorescence staining were performed to analyze immune cell infiltration in the tumor microenvironment.Results·Database analyses,qRT-PCR,and Western blotting confirmed that some human melanoma cell lines exhibited high STING mRNA expression but low STING protein levels.YM201636 significantly increased STING protein expression in YUMM1.7 cells(P<0.001).Combined treatment with YM201636 and diABZI significantly enhanced phosphorylation of TBK1 and IRF3(P<0.05),indicating effective activation of the STING signaling pathway.This combination also promoted the expression of type Ⅰ interferons(P<0.001)and enhanced tumor cell killing in vitro.In vivo,the combination therapy markedly suppressed melanoma growth compared to monotherapy.Immune profiling of the tumor microenvironment revealed significantly increased infiltration of CD4⁺ T cells and CD8⁺ T cells in the combination treatment group(P<0.05).Conclusion·The PIKfyve inhibitor YM201636 could restore STING protein expression in STING-deficient melanoma and enhance the antitumor efficacy of the STING agonist diABZI,offering a promising therapeutic strategy for tumors with defective STING signaling.关键词
黑色素瘤/STING激动剂/PIKfyve抑制剂Key words
melanoma/STING agonist/PIKfyve inhibitor分类
医药卫生引用本文复制引用
杨小雨,黄蕊,吴以加,张哲,房燕,沈键锋..PIKfyve抑制剂靶向治疗STING通路缺陷黑色素瘤的机制研究[J].上海交通大学学报（医学版）,2025,45(9):1126-1137,12.基金项目
国家重点研发计划(2021YFC2701103) （2021YFC2701103）
上海交通大学医学院"双百人"项目(20191817). National Key R&D Program of China(2021YFC2701103) （20191817）
"Two-hundred Talents"Program of Shanghai Jiao Tong University School of Medicine(20191817). （20191817）
