Laboratory of Viral Immunopathology, Unit of Infectious Diseases and Hepatology, Azienda Ospedaliero-Universitaria di Parma, Parma, Italy.
Department of Medicine and Surgery, University of Parma, Via Gramsci 14, 43126, Parma, Italy.
Cancer Immunol Immunother. 2020 Aug;69(8):1589-1603. doi: 10.1007/s00262-020-02561-4. Epub 2020 Apr 19.
Functional rescue of NK-cells in solid tumors represents a central aim for new immunotherapeutic strategies. We have conducted a genomic, phenotypic and functional analysis of circulating NK-cells from patients with HCV-related liver cirrhosis and hepatocellular carcinoma. NK-cells were sorted from patients with HCC or liver cirrhosis and from healthy donors. Comparative mRNA gene expression profiling by whole-human-genome microarrays of sorted NK-cells was followed by phenotypic and functional characterization. To further identify possible mediators of NK-cell dysfunction, an in vitro model using media conditioned with patients' and controls' plasma was set up. Metabolic and cell motility defects were identified at the genomic level. Dysregulated gene expression profile has been translated into reduced cytokine production and degranulation despite a prevalent phenotype of terminally differentiated NK-cells. NKG2D-downregulation, high SMAD2 phosphorylation and other phenotypic and molecular alterations suggested TGF-β as possible mediator of this dysfunction. Blocking TGF-β could partially restore functional defects of NK-cells from healthy donors, exposed to TGF-β rich HCC patients' plasma, suggesting that TGF-β among other molecules may represent a suitable target for immunotherapeutic intervention aimed at NK-cell functional restoration. By an unbiased approach, we have identified energy metabolism and cell motility defects of circulating NK-cells as main mechanisms responsible for functional NK-cell impairment in patients with hepatocellular carcinoma. This opens the way to test different approaches to restore NK-cell response in these patients.
在实体肿瘤中功能性 NK 细胞的恢复是新免疫治疗策略的主要目标。我们对与 HCV 相关的肝硬化和肝细胞癌患者的循环 NK 细胞进行了基因组、表型和功能分析。我们从 HCC 或肝硬化患者以及健康供体中分离 NK 细胞。通过全人类基因组微阵列对分选的 NK 细胞进行比较 mRNA 基因表达谱分析,然后进行表型和功能特征分析。为了进一步鉴定 NK 细胞功能障碍的可能介质,我们建立了使用患者和对照血浆条件培养基的体外模型。在基因组水平上鉴定到代谢和细胞迁移缺陷。尽管 NK 细胞表现出终末分化的表型,但失调的基因表达谱已转化为细胞因子产生和脱颗粒减少。NKG2D 下调、SMAD2 高磷酸化和其他表型和分子改变表明 TGF-β 可能是这种功能障碍的介导物。阻断 TGF-β可部分恢复暴露于富含 TGF-β 的 HCC 患者血浆的健康供体 NK 细胞的功能缺陷,这表明 TGF-β 等分子可能是针对旨在恢复 NK 细胞功能的免疫治疗干预的合适靶点。通过一种无偏见的方法,我们鉴定了循环 NK 细胞的能量代谢和细胞迁移缺陷,作为 HCC 患者功能性 NK 细胞损伤的主要机制。这为在这些患者中恢复 NK 细胞反应的不同方法提供了可能。