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.
CELL INTELLIGENCE · 肿瘤细胞治疗研究
肿瘤细胞治疗研究
用于鉴定抗体依赖性细胞毒性调节化合物的高内涵筛选检测
英文原题：High-Content Screening Assay for the Identification of Antibody-Dependent Cellular Cytotoxicity Modifying Compounds.
查看英文原题
High-Content Screening Assay for the Identification of Antibody-Dependent Cellular Cytotoxicity Modifying Compounds.
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中文摘要
采用抗原特异性抗体或免疫检查点抑制剂的免疫疗法已改变乳腺癌治疗格局。表达人表皮生长因子受体 2（HER2）的乳腺癌细胞可由抗 HER2 抗体 trastuzumab 靶向。抗体依赖性细胞毒作用（ADCC）是 HER2 抗肿瘤作用的重要机制之一。与癌细胞结合的 trastuzumab 可被 ADCC 效应细胞（如自然杀伤（NK）细胞、巨噬细胞和粒细胞）的 Fc 受体识别，触发这些免疫细胞的细胞毒活性，最终导致癌细胞死亡。
本研究拟开发一种基于成像的 ADCC 定量检测方法，通过高内涵筛选发现新的 ADCC 调节化合物。检测中，将 HER2 过表达的 JIMT-1 乳腺癌细胞与 NK-92 细胞在 trastuzumab 存在下共培养，并通过自动显微镜和定量图像分析测定靶细胞死亡。根据 EGFP 荧光区分靶细胞与效应细胞。本文展示了如何用该检测筛选化合物库，以发现 ADCC 调节药物。研究使用从实验室现有试剂中随机选取的精细化学品建立化合物库测试板，并额外纳入 3 种预期会影响 NK 细胞迁移和脱颗粒的微管去稳定剂（colchicine、vincristine 和 podophyllotoxin）。测试筛选将全部 3 种阳性对照化合物识别为命中，证明该方法适用于从化学库中筛选 ADCC 调节药物。利用该检测开展化合物库筛选，可能发现能够增强 ADCC 的化合物，并将其作为抗癌免疫治疗患者的辅助治疗药物。
此外，该方法还可识别癌症患者因其他适应证服用的治疗药物可能产生的非预期 ADCC 抑制副作用。
展开英文摘要原文
Immunotherapy with antigen-specific antibodies or immune checkpoint inhibitors has revolutionized the therapy of breast cancer. Breast cancer cells expressing the epidermal growth factor receptor HER2 can be targeted by the anti-HER-2 antibody trastuzumab.
Antibody-dependent cellular cytotoxicity (ADCC) is an important mechanism implicated in the antitumor action of HER-2. Trastuzumab bound to cancer cells can be recognized by the Fc receptors of ADCC effector cells (e. g. , natural killer (NK) cells, macrophages, and granulocytes), triggering the cytotoxic activity of these immune cells leading to cancer cell death.
We set out to develop an image-based assay for the quantification of ADCC to identify novel ADCC modulator compounds by high-content screening. In the assay, HER2 overexpressing JIMT-1 breast cancer cells are co-cultured with NK-92 cells in the presence of trastuzumab, and target cell death is quantified by automated microscopy and quantitative image analysis. Target cells are distinguished from effector cells based on their EGFP fluorescence.
We show how compound libraries can be tested in the assay to identify ADCC modulator drugs. For this purpose, a compound library test plate was set up using randomly selected fine chemicals off the lab shelf. Three microtubule destabilizing compounds (colchicine, vincristine, podophyllotoxin) expected to interfere with NK cell migration and degranulation were also included in the test library.
The test screen identified all three positive control compounds as hits proving the suitability of the method to identify ADCC-modifying drugs in a chemical library. With this assay, compound library screens can be performed to identify ADCC-enhancing compounds that could be used as adjuvant therapeutic agents for the treatment of patients receiving anticancer immunotherapies.
In addition, the method can also be used to identify any undesirable ADCC-inhibiting side effects of therapeutic drugs taken by cancer patients for different indications.
论文信息
- 作者
- Guti E、Bede ÁM、Váróczy C、Hegedűs C、Demény MÁ、Virág L
- 第一作者单位
- Department of Medical Chemistry, Faculty of Medicine, University of Debrecen; Doctoral School of Molecular Medicine, University of Debrecen.
- 通讯作者单位
- Department of Medical Chemistry, Faculty of Medicine, University of Debrecen; ELKH-DE Cell Biology and Signaling Research Group; lvirag@med.unideb.hu.
- 文献类型
- 音视频资料 · 非美国政府资助研究
- 期刊
- Journal of visualized experiments : JoVE2023 Aug 18
- 原文标识
- PubMed 37607075 · DOI 10.3791/64485