Journal of Shanghai Jiao Tong University (Medical Science) >

2025 , Vol. 45 >Issue 9: 1126 - 1137

DOI: https://doi.org/10.3969/j.issn.1674-8115.2025.09.005

Basic research

Mechanistic study of targeting melanoma with STING pathway deficiencies via PIKfyve inhibitor

  • YANG Xiaoyu ,
  • HUANG Rui ,
  • WU Yijia ,
  • ZHANG Zhe ,
  • FANG Yan ,
  • SHEN Jianfeng
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  • Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology, Shanghai 200011, China
SHEN Jianfeng, E-mail: jfshen@shsmu.edu.cn.
FANG Yan, E-mail: yanyan2021fang@sjtu.edu.cn.

Received date: 2025-03-30

  Accepted date: 2025-04-30

  Online published: 2025-09-30

Supported by

National Key R&D Program of China(2021YFC2701103);“Two-hundred Talents” Program of Shanghai Jiao Tong University School of Medicine(20191817)

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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 STINGpathway-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.

Key words： melanoma; STING agonist; PIKfyve inhibitor

Cite this article

YANG Xiaoyu , HUANG Rui , WU Yijia , ZHANG Zhe , FANG Yan , SHEN Jianfeng . Mechanistic study of targeting melanoma with STING pathway deficiencies via PIKfyve inhibitor[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2025 , 45(9) : 1126 -1137 . DOI: 10.3969/j.issn.1674-8115.2025.09.005

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