PO.TB10.16 · 肿瘤生物学
通过破坏nNOS/NO与COX-2之间的相互作用抑制干扰素-gamma刺激的黑色素瘤进展
Inhibiting interferon-gamma-stimulated melanoma progression by disrupting the crosstalk between nNOS/NO and COX-2
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摘要 Abstract
中文摘要
黑色素瘤肿瘤微环境中的干扰素gamma（IFN-gamma）发挥相反的作用，既协调促肿瘤活性又协调抗癌免疫应答。我们此前的研究证明了神经元型一氧化氮合酶（nNOS）在IFN-gamma刺激的黑色素瘤进展中的作用。然而，其潜在机制尚未得到充分阐明。我们对患者和细胞蛋白质组数据进行生物信息学分析，以鉴定与黑色素瘤中IFN-gamma处理相关的目标蛋白。我们的组学分析显示，COX-2的诱导可显著预测黑色素瘤细胞中的IFN-gamma处理。在IFN-gamma存在下，PGE 2进一步增强PD-L1表达并放大nNOS的诱导，从而增加细胞内NO水平。与选择性COX-2抑制剂塞来昔布联合处理可有效减弱PGE 2诱导的这些变化。使用选择性小分子抑制剂（HH044）阻断nNOS可有效抑制IFN-gamma诱导的黑色素瘤细胞中PGE 2产生和COX-2表达。STAT3抑制剂napabucasin在IFN-gamma存在和不存在的情况下均抑制COX-2表达。体内实验中，HH044处理显著降低了人黑色素瘤异种移植小鼠模型（A375/Nu/Nu）中的肿瘤PGE 2水平。在同基因小鼠黑色素瘤模型（Cloudman S91/DBA/2）中，HH044与塞来昔布联合处理产生最强的肿瘤抑制，将肿瘤体积降至对照的19%，而单用HH044或塞来昔布分别为38%和52%。此外，转录组分析显示HH044处理后参与基质重塑和转移的基因发生显著变化。离体将人PBMC与黑色素瘤细胞共培养可抑制T细胞活化，在IFN-gamma存在和不存在时均减少分泌IL-2的T细胞。来自相当比例供体（64%）的PBMC经HH044预处理后被重新激活，与黑色素瘤细胞共孵育后IL-2+ T细胞显著增加。我们的研究揭示了一个正反馈环路，将nNOS介导的NO信号与COX-2/PGE 2信号轴相连，从而进一步增强IFN-gamma的促肿瘤活性。用选择性抑制剂破坏nNOS/NO与COX-2之间的相互作用，可能通过调节肿瘤免疫微环境有效抑制黑色素瘤肿瘤生长。
查看英文原文 English abstract
Interferon gamma (IFN-gamma) in the melanoma tumor microenvironment plays opposing roles, orchestrating both pro-tumorigenic activity and anticancer immune responses. Our previous studies demonstrated the role of neuronal nitric oxide synthase (nNOS) in IFN-gamma-stimulated melanoma progression. However, the underlying mechanism has not been well defined. Bioinformatic analysis of patient and cellular proteomic data was conducted to identify proteins of interest associated with IFN-gamma treatment in melanoma. Our omics analysis revealed that the induction of COX-2 was significantly predictive of IFN-gamma treatment in melanoma cells. In the presence of IFN-gamma, PGE 2 further enhanced PD-L1 expression and amplified the induction of nNOS, which increased intracellular NO levels. Cotreatment with celecoxib, a selective COX-2 inhibitor, effectively diminished these changes induced by PGE 2 . nNOS blockade using a selective small molecule inhibitor (HH044) efficiently inhibited IFN-gamma-Induced PGE 2 production and COX-2 expression in melanoma cells. STAT3 inhibitor napabucasin also inhibited COX-2 expression both in the presence and absence of IFN-gamma. In vivo , HH044 treatment significantly reduced tumor PGE 2 levels in a human melanoma xenograft mouse model (A375/Nu/Nu). Combination treatment with HH044 and celecoxib yielded the greatest tumor suppression in the syngeneic murine melanoma model (Cloudman S91/DBA/2), reducing tumor volume to 19% of control, compared to 38% and 52% with HH044 or celecoxib alone, respectively. Furthermore, transcriptomic analysis revealed significant changes in genes involved in matrix remodeling and metastasis after HH044 treatment. Ex vivo co-culturing human PBMCs with melanoma cells inhibited T cell activation, decreasing IL-2-secreting T cells in the presence and absence of IFN-gamma. PBMCs from a significant portion of donors (64%) were reactivated by HH044 pretreatment, displaying a significant increase in IL-2+ T cells after coincubation with melanoma cells.Our study reveals a positive feedback loop linking nNOS-mediated NO signaling to the COX-2/PGE 2 signaling axis in melanoma, thereby further enhancing the pro-tumorigenic activity of IFN-gamma. Disrupting crosstalk between nNOS/NO and COX-2 with selective inhibitors effectively suppressed melanoma tumor growth, possibly by modulating the tumor immune microenvironment.
利益披露 Disclosure
A. Patel, None..
K. A. Lozada, None..
M. Roosan, None..
B. Syed, None..
J. Totonchy, None..
A. Awasthi, None..
R. B. Silverman, None.
S. Yang,
Merck, Inc ).
Amgen ).