Jia Tengfei, Guo Yingxi, Cheng Xin Meng, Zhou Zeyang, Xu Xiaojiang, Liu Hebin, Yang Xiaodong
Institutes of Biology and Medical Sciences, Soochow University, Suzhou, China.
Department of Gastrointestinal Surgery, The Second Affiliated Hospital of Soochow University, Suzhou, China.
Clin Transl Med. 2025 Aug;15(8):e70425. doi: 10.1002/ctm2.70425.
Colorectal cancer (CRC) ranks among the most prevalent malignant tumours of the digestive system globally and is associated with unfavourable survival outcomes. The exhaustion of CD8⁺ T cells serves a crucial role in facilitating tumour immune escape. Yet, the dynamic evolution of CD8⁺ T cell exhaustion and its impact on clinical prognosis across TNM (tumour-node-metastasis) stages in CRC remains incompletely characterized.
Tumour and adjacent tissues (20 samples total) from 6 CRC patients spanning diverse TNM stages were analyzed using integrated single-cell transcriptomic profiling (scRNA-seq), single-cell T cell receptor/B cell receptor sequencing (scVDJ-seq), and spatial transcriptomics. T cell exhaustion markers, immune clonality, gene expression profiles, and the spatial distribution of both tumour cells and immune cells were systematically profiled. Functional enrichment and intercellular communication analyses were conducted. Key findings were validated using immunofluorescence and public datasets.
Our results illustrate how advancing TNM stages in CRC shape CD8⁺ T cell exhaustion through divergent TNFRSF18/CXCL13 dynamics and ribosomal stemness. TNFRSF18 expression was notably higher in T cells infiltrating tumour tissues relative to their counterparts in adjacent non-tumorous areas, with high-expressing CD8⁺ T cells exhibiting marked exhaustion features. During CRC progression, TNM-stage-driven remodelling of the tumour microenvironment (TME) induced progressive CD8⁺ T cell exhaustion marked by declining TNFRSF18 and rising CXCL13 expression in tumour-infiltrating T cells elevation of both markers in the tumour compared with adjacent tissues. Moreover, we show that tumour cells displayed elevated expression of stemness-associated ribosomal genes (RPS7, RPL8, RPL30), peaking at stage T4, which correlated with poor prognosis and immune escape.
This integrative multi-omics study uncovers CD8⁺ T cell exhaustion dynamics and ribosomal stemness-mediated immune evasion across CRC progression. CXCL13, TNFRSF18, and ribosomal proteins (RPS7/RPL8/RPL30) are identified as novel biomarkers with direct prognostic value and therapeutic relevance, providing therapeutic targets for precision immunotherapy in CRC.
Multi-omics analysis reveals dynamic CD8 T cell exhaustion patterns across CRC samples with different TNM stages. TNFRSF18 is highly expressed in exhausted tumour-infiltrating CD8 T cells and declines with disease progression. Ribosomal stemness in tumour cells promotes immune evasion by impairing TNF-mediated CD8 T cell function.
结直肠癌（CRC）是全球消化系统中最常见的恶性肿瘤之一，且与不良生存结果相关。CD8⁺T细胞耗竭在促进肿瘤免疫逃逸中起关键作用。然而，CRC中CD8⁺T细胞耗竭的动态演变及其对TNM（肿瘤-淋巴结-转移）各阶段临床预后的影响仍未完全明确。
对6例处于不同TNM阶段的CRC患者的肿瘤及相邻组织（共20个样本）进行综合单细胞转录组分析（scRNA-seq）、单细胞T细胞受体/B细胞受体测序（scVDJ-seq）和空间转录组学分析。系统分析T细胞耗竭标志物、免疫克隆性、基因表达谱以及肿瘤细胞和免疫细胞的空间分布。进行功能富集和细胞间通讯分析。利用免疫荧光和公共数据集对关键发现进行验证。
我们的结果表明，CRC中TNM分期的进展如何通过不同的TNFRSF18/CXCL13动态变化和核糖体干性塑造CD8⁺T细胞耗竭。相对于相邻非肿瘤区域的T细胞，肿瘤浸润T细胞中TNFRSF18表达显著更高，高表达的CD8⁺T细胞表现出明显的耗竭特征。在CRC进展过程中，TNM分期驱动的肿瘤微环境（TME）重塑诱导肿瘤浸润T细胞中TNFRSF18表达下降和CXCL13表达上升，从而导致CD8⁺T细胞逐渐耗竭，与相邻组织相比，肿瘤中这两种标志物均升高。此外，我们发现肿瘤细胞中干性相关核糖体基因（RPS7、RPL8、RPL30）表达升高，在T4期达到峰值，这与预后不良和免疫逃逸相关。
这项综合多组学研究揭示了CRC进展过程中CD8⁺T细胞耗竭动态变化以及核糖体干性介导的免疫逃逸。CXCL13、TNFRSF18和核糖体蛋白（RPS7/RPL8/RPL30）被确定为具有直接预后价值和治疗相关性的新型生物标志物，为CRC精准免疫治疗提供了治疗靶点。
多组学分析揭示了不同TNM分期的CRC样本中CD8 T细胞耗竭的动态模式。TNFRSF18在耗竭的肿瘤浸润CD8 T细胞中高表达，并随疾病进展而下降。肿瘤细胞中的核糖体干性通过损害TNF介导的CD8 T细胞功能促进免疫逃逸。
