PKCα/ZFP64/CSF1 轴重置肿瘤微环境并促进肝细胞癌中的抗 PD1 耐

StephenW

PKCα/ZFP64/CSF1 轴重置肿瘤微环境并促进肝细胞癌中的抗 PD1 耐药性

    魏传元#
    朱梦轩#
    张鹏飞#
    范家斌
    艾无克
    贾凡
   
开放存取发布时间：2022 年 2 月 23 日 DOI：https://doi.org/10.1016/j.jhep.2022.02.019


强调

    •
    ZFP64 在抗 PD1 抗性 HCC 中经常上调。
    •
    PKCα/ZFP64/CSF1 轴对于触发免疫逃避和抗 PD1 耐受至关重要。
    •
    Gö6976 和 lenvatinib 通过阻断 PKCα/ZFP64/CSF1 轴克服抗 PD1 耐药性。
    •
    Gö6976 联合抗 PD1 可能是一种有效的 HCC 治疗新策略。

背景与目标
尽管治疗取得了显着进展，但大多数肝细胞癌 (HCC) 患者对抗程序性细胞死亡 1 (anti-PD1) 治疗反应不佳。迫切需要更深入地了解 HCC 对这种疗法的耐受机制。
方法
我们进行了二代测序、多重免疫荧光和双色免疫组织化学，并构建了原位 HCC 异种移植肿瘤模型，以确定与抗 PD1 耐受性相关的关键基因。使用自发致瘤转基因小鼠模型、体外共培养系统、大规模细胞术和多重免疫荧光来探索锌指蛋白 64 (ZFP64) 在肿瘤进展和免疫逃逸中的生物学功能。 RNA测序、染色质免疫沉淀测序和质谱等分子和生化策略被用来深入了解ZFP64的潜在机制。
结果
我们发现 ZFP64 在抗 PD1 耐药 HCC 患者的肿瘤组织中经常上调。升高的 ZFP64 通过将巨噬细胞极化转变为替代激活表型 (M2) 并促进抑制性肿瘤微环境来驱动抗 PD1 抗性。从机制上讲，我们主要证明蛋白激酶 C α (PKCα) 在 S226 处直接磷酸化 ZFP64，导致其核转位和巨噬细胞集落刺激因子 (CSF1) 的转录激活。 HCC 衍生的 CSF1 将巨噬细胞转化为 M2 表型，以驱动免疫逃逸和抗 PD1 耐受。值得注意的是，蛋白激酶抑制剂 Gö6976 和多激酶抑制剂 lenvatinib 通过阻断 PKCα/ZFP64/CSF1 轴来重置肿瘤微环境并恢复对抗 PD1 的敏感性。
结论
我们提出 PKCα/ZFP64/CSF1 轴对于触发免疫逃避和抗 PD1 耐受至关重要。用 Gö6976 或 lenvatinib 抑制该轴可克服 HCC 中的抗 PD1 耐药性。
总结
尽管治疗取得了显着进展，但大多数肝细胞癌患者对抗 PD1 疗法（一种免疫疗法）反应不佳。迫切需要更深入地了解这种疗法的耐受机制。在此，我们揭示了一种先前未探索的与肿瘤进展、巨噬细胞极化和抗 PD1 抗性相关的机制，并为抗 PD1 联合治疗提供了一个有吸引力的新靶点，这可能使肝细胞癌患者受益。

StephenW

PKCα/ZFP64/CSF1 axis resets the tumor microenvironment and fuels anti-PD1 resistance in hepatocellular carcinoma

    Chuan-Yuan Wei #
    Meng-Xuan Zhu #
    Peng-Fei Zhang #
    Jia-Bin Fan
    Ai-Wu Ke
    Jia Fan
   
Open AccessPublished:February 23, 2022DOI:https://doi.org/10.1016/j.jhep.2022.02.019


Highlights

    •
    ZFP64 is frequently upregulated in anti-PD1 resistant HCC.
    •
    PKCα/ZFP64/CSF1 axis is critical for triggering immune evasion and anti-PD1 tolerance.
    •
    Gö6976 and lenvatinib overcome anti-PD1 resistance by blocking the PKCα/ZFP64/CSF1 axis.
    •
    Gö6976 combined with anti-PD1 could be an effective new strategy in HCC therapy.

Background & Aims
Despite remarkable advances in treatment, most patients with hepatocellular carcinoma (HCC) respond poorly to anti-programmed cell death 1 (anti-PD1) therapy. A deeper insight into the tolerance mechanism of HCC against this therapy is urgently needed.
Methods
We performed next-generation sequencing, multiplex immunofluorescence, and dual-color immunohistochemistry and constructed an orthotopic HCC xenograft tumor model to identify the key gene associated with anti-PD1 tolerance. A spontaneously tumorigenic transgenic mouse model, an in vitro coculture system, mass cytometry, and multiplex immunofluorescence were used to explore the biological function of zinc finger protein 64 (ZFP64) on tumor progression and immune escape. Molecular and biochemical strategies like RNA-sequencing, chromatin immunoprecipitation-sequencing and mass spectrometry were used to gain insight into the underlying mechanisms of ZFP64.
Results
We showed that ZFP64 is frequently upregulated in tumor tissues from patients with anti-PD1-resistant HCC. Elevated ZFP64 drives anti-PD1 resistance by shifting macrophage polarization toward an alternative activation phenotype (M2) and fostering an inhibitory tumor microenvironment. Mechanistically, we primarily demonstrated that protein kinase C alpha (PKCα) directly phosphorylates ZFP64 at S226, leading to its nuclear translocation and the transcriptional activation of macrophage colony-stimulating factor (CSF1). HCC-derived CSF1 transforms macrophages to the M2 phenotype to drive immune escape and anti-PD1 tolerance. Notably, Gö6976, a protein kinase inhibitor, and lenvatinib, a multi-kinase inhibitor, reset the tumor microenvironment and restore sensitivity to anti-PD1 by blocking the PKCα/ZFP64/CSF1 axis.
Conclusions
We propose that the PKCα/ZFP64/CSF1 axis is critical for triggering immune evasion and anti-PD1 tolerance. Inhibiting this axis with Gö6976 or lenvatinib overcomes anti-PD1 resistance in HCC.
Lay summary
Despite remarkable treatment progress, most patients with hepatocellular carcinoma respond poorly to anti-PD1 therapy (a type of immunotherapy). A deeper insight into the tolerance mechanisms to this therapy is urgently needed. Herein, we unravel a previously unexplored mechanism linking tumor progression, macrophage polarization, and anti-PD1 resistance, and offer an attractive novel target for anti-PD1 combination therapy, which may benefit patients with hepatocellular carcinoma.

StephenW

http://www.journal-of-hepatology.eu/article/S016882782200126X/pdf