Department of Lymphatic Medical Oncology, The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences, Hangzhou, 310022 Zhejiang, China.
Cancer Institute (Key Laboratory of Cancer Prevention & Intervention, National Ministry of Education, Provincial Key Laboratory of Molecular Biology in Medical Sciences), The Second Affiliated Hospital Zhejiang University School of Medicine, Hangzhou, 310009 Zhejiang, China.
Biomed Res Int. 2021 Jun 19;2021:5514726. doi: 10.1155/2021/5514726. eCollection 2021.
Our purpose was to characterize distinct molecular subtypes of diffuse large B cell lymphoma (DLBCL) patients treated with rituximab-CHOP (R-CHOP).
Two gene expression datasets of R-CHOP-treated DLBCL patients were downloaded from GSE10846 ( = 233, training set) and GSE31312 ( = 470, validation set) datasets. Cluster analysis was presented via the ConsensusClusterPlus package in R. Using the limma package, differential expression analysis was utilized to identify feature genes. Kaplan-Meier survival analysis was presented to compare the differences in the prognosis between distinct molecular subtypes. Correlation between molecular subtypes and clinical features was analyzed. Based on the sets of highly expressed genes, biological functions were explored by gene set enrichment analysis (GSEA). Several feature genes were validated in the molecular subtypes via qRT-PCR and western blot.
DLBCL samples were clustered into two molecular subtypes. Samples in subtype I displayed poorer overall survival time in the training set ( < 0.0001). Consistently, patients in subtype I had shorter overall survival ( = 0.0041) and progression-free survival time ( < 0.0001) than those in subtype II. Older age, higher stage, and higher international prognostic index (IPI) were found in subtype I. In subtype I, T cell activation, lymphocyte activation, and immune response were distinctly enriched, while cell adhesion, migration, and motility were significantly enriched in subtype II. T cell exhaustion-related genes including TIM3 ( < 0.001), PD-L1 ( < 0.0001), LAG3 ( < 0.0001), CD160 ( < 0.001), and CD244 ( < 0.001) were significantly highly expressed in subtype I than subtype II.
Two molecular subtypes were constructed in DLBCL, which were characterized by different clinical outcomes and molecular mechanisms. Our findings may offer a novel insight into risk stratification and prognosis prediction for DLBCL patients.
本研究旨在分析接受利妥昔单抗联合环磷酰胺、多柔比星、长春新碱和泼尼松（R-CHOP）治疗的弥漫性大 B 细胞淋巴瘤（DLBCL）患者的不同分子亚型特征。
从 GSE10846（n = 233，训练集）和 GSE31312（n = 470，验证集）数据集下载了两个接受 R-CHOP 治疗的 DLBCL 患者的基因表达数据集。使用 R 中的 ConsensusClusterPlus 包进行聚类分析。使用 limma 包进行差异表达分析，以鉴定特征基因。通过 Kaplan-Meier 生存分析比较不同分子亚型之间的预后差异。分析分子亚型与临床特征的相关性。基于高表达基因集，通过基因集富集分析（GSEA）探索生物学功能。通过 qRT-PCR 和 Western blot 在分子亚型中验证了几个特征基因。
将 DLBCL 样本聚类为两个分子亚型。在训练集中，亚型 I 的样本总生存时间更差（<0.0001）。一致地，与亚型 II 相比，亚型 I 的患者总生存时间（=0.0041）和无进展生存时间（<0.0001）更短。在亚型 I 中发现年龄较大、分期较高和国际预后指数（IPI）较高。在亚型 I 中，T 细胞激活、淋巴细胞激活和免疫反应明显富集，而细胞黏附、迁移和运动在亚型 II 中明显富集。T 细胞耗竭相关基因，包括 TIM3（<0.001）、PD-L1（<0.0001）、LAG3（<0.0001）、CD160（<0.001）和 CD244（<0.001），在亚型 I 中明显高于亚型 II。
在 DLBCL 中构建了两个分子亚型，其特征在于不同的临床结局和分子机制。我们的研究结果可能为 DLBCL 患者的风险分层和预后预测提供新的见解。
