RNF43 p.G659fs 通过 PI3K/AKT/mTOR 信号通路和 HLA-E 上调导致 MSI-high 结直肠癌中 NK 细胞功能障碍
RNF43 p.G659fs leads to natural killer cell dysfunction in MSI-high colorectal cancer through PI3K/AKT/mTOR signaling and HLA-E up-regulation.
肿瘤细胞治疗研究
英文原题:Bulk RNA sequencing reveals the comprehensive genetic characteristics of human cord blood-derived natural killer cells.
Bulk RNA sequencing reveals the comprehensive genetic characteristics of human cord blood-derived natural killer cells.
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我们展示了人类 CBNKCs 的特征。使用 CBNKCs 的细胞疗法有望用于治疗 GBM。
先天免疫细胞在肿瘤免疫治疗中具有重要作用。NK 细胞(NKC)也被归类为先天免疫细胞,能够控制肿瘤生长和转移扩散。胶质母细胞瘤(GBM)是成人中最常见且最具侵袭性的原发性脑肿瘤。基于NKC的免疫治疗是治疗GBM的一种有前景的策略。我们此前报道了一种无饲养层扩增系统,可从人脐带血(CB)中获得大规模高纯度且具有细胞毒性的NKC。在本研究中,我们对由人CB生成的NKC(CBNKC)与人外周血(PB)生成的NKC(PBNKC)进行了全面的基因组学分析比较。
冻存的T细胞无CB单核细胞在抗NKp46和抗CD16抗体固定化条件下,与重组人白细胞介素(rhIL)-18和rhIL-2共培养。扩增14天后,提取CBNKCs或PBNKCs的总RNA,并进行转录组分析,以确定它们的相似性和差异。我们还检测了CBNKC和PBNKC对GBM细胞系的活性。
差异表达基因分析显示,与PBNKCs相比,CBNKCs中部分NK激活性和抑制性受体显著下调。此外,CBNKCs中与抗凋亡和增殖相关的基因上调。富集分析确定,与免疫应答和细胞因子相关的基因集在CBNKCs中富集。基因集富集分析表明,免疫应答通路在CBNKCs中上调。使用基于阻抗的细胞分析仪进行的细胞毒性实验显示,与PBNKCs相比,CBNKCs增强了NKC介导的对GBM细胞的细胞毒性。
Frozen T cell-free CB mononuclear cells were cultured with recombinant human interleukin (rhIL)-18 and rhIL-2 in anti-NKp46 and anti-CD16 antibody immobilization settings. After 14-day expansion, the total RNA of the CBNKCs or PBNKCs was extracted and transcriptomic analyses was performed to determine their similarities and differences. We also examined CBNKC and PBNKC activity against a GBM cell line.
Differential expression gene analysis revealed that some NK activating and inhibitory receptors were significantly downregulated in the CBNKCs compared to PBNKCs. Furthermore, genes related to anti-apoptosis and proliferation were upregulated in the CBNKCs. Enrichment analysis determined that the gene sets related to immune response and cytokines were enriched in the CBNKCs. Gene set enrichment analysis demonstrated that the immune response pathway was upregulated in the CBNKCs. Cytotoxic assays using impedance-based cell analyzer revealed that the CBNKCs enhanced NKC-mediated cytotoxicity on GBM cells as compared to the PBNKCs.
We demonstrated the characteristics of human CBNKCs. Cell-based therapy using the CBNKCs is promising for treating GBM.
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