Department of Pulmonary Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, Huaihai West Road No.241, Shanghai, 200030, China.
Department of Critical Care, Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
Cancer Immunol Immunother. 2021 Aug;70(8):2261-2274. doi: 10.1007/s00262-021-02848-0. Epub 2021 Jan 27.
High expression of PD-L1 predicts PD-1/PD-L1 inhibitor benefit, meanwhile a few PD-L1-negative patients still benefit from these drugs. In this study, we aimed to explore the underlying cellular and molecular characteristics via single-cell sequencing. Before and after treatment with Pembrolizumab, peripheral blood mononuclear cells (PBMCs) were isolated via Ficoll gradient. Thereafter, single-cell RNA sequencing was performed, and clinical significance was validated with The Cancer Genome Atlas (TCGA) cohort. All 3423 cells of 16 clusters were classified into eight cell types, including NKG7+ T, NKG7+ NK, Naïve T, CDC1C+ dendritic cells, CD8+ T cells, B cells, macrophages and erythrocytes. Cell proportion, the clinical significance of differentially expressed genes and significant pathways of NKG7+ T, NKG7+ NK, Naïve T and CD8+ T cells were analyzed. Ubiquitin-mediated proteolysis/cell cycle/natural killer cell-mediated cytotoxicity were identified as PD-1 blockage-responsive pathways in NKG7+ NK cells. Apoptosis/Th1 and Th2 cell differentiation were proposed as Pembrolizumab-affected pathways in NKT cells. In gene level, ID2, PIK3CD, UQCR10, MATK, MZB1, IL7R and TRGC2 showed a significant correlation with PD-1 expression after TCGA dataset validation, which could possess potential as predictive markers for patients with PD-L1-negative lung squamous cell carcinoma who can benefit from Pembrolizumab.
高表达的 PD-L1 预测 PD-1/PD-L1 抑制剂的获益，同时少数 PD-L1 阴性患者仍能从这些药物中获益。在这项研究中，我们旨在通过单细胞测序来探索潜在的细胞和分子特征。在接受 Pembrolizumab 治疗前后，通过 Ficoll 梯度分离外周血单核细胞（PBMCs）。然后进行单细胞 RNA 测序，并通过 The Cancer Genome Atlas（TCGA）队列验证其临床意义。在 16 个簇的 3423 个细胞中，将所有细胞分为 8 种细胞类型，包括 NKG7+T、NKG7+NK、幼稚 T、CDC1C+树突状细胞、CD8+T 细胞、B 细胞、巨噬细胞和红细胞。分析细胞比例、差异表达基因的临床意义以及 NKG7+T、NKG7+NK、幼稚 T 和 CD8+T 细胞的显著通路。泛素介导的蛋白水解/细胞周期/自然杀伤细胞介导的细胞毒性被鉴定为 NKG7+NK 细胞中 PD-1 阻断反应途径。细胞凋亡/Th1 和 Th2 细胞分化被认为是 NKT 细胞中 Pembrolizumab 作用的途径。在基因水平上，ID2、PIK3CD、UQCR10、MATK、MZB1、IL7R 和 TRGC2 在 TCGA 数据集验证后与 PD-1 表达呈显著相关，它们可能作为 PD-L1 阴性肺鳞癌患者从 Pembrolizumab 中获益的潜在预测标志物。
