Wang Sheng, Ye Geni, Xu Xiaoling, Lin Yuning, Li Xiaobo, Zhang Zhang, Qi Ming, Tan Lin, Cheng Minjing, Zhang Haishan, Pan Jinghua, Lin Changwei, Huang Dandan, Deng Rong, Li Xiaomei, Wang Guangsuo, Qiu Shenghui, Chu Xiaodong, Chen Yuhong, Zeng Huhu, Zhang Junqiu, Wang Chenran, Shuai Hanlin, Shi Changzheng, Yuan Xia, Cao Yihai, Chen Minfeng, Huang Maohua, Ye Wencai, Zhang Dongmei
State Key Laboratory of Bioactive Molecules and Druggability Assessment, Guangdong Basic Research Center of Excellence for Natural Bioactive Molecules and Discovery of Innovative Drugs, Jinan University, Guangzhou, Guangdong, China.
College of Pharmacy, Jinan University, Guangzhou, Guangdong, China.
Gut. 2025 Jul 23. doi: 10.1136/gutjnl-2024-334618.
Circulating tumour cell (CTC)-neutrophil clusters represent a key driver and a hallmark of tumour metastasis; however, efficient approaches for their elimination are still lacking.
This study sought to elucidate the location and mechanisms of CTC-neutrophil cluster formation and develop more effective antimetastasis strategies.
Immunofluorescence staining of clinical colorectal cancer (CRC) samples was performed to identify the location of CTC-neutrophil clusters. The correlation between the expression of nicotinamide N-methyltransferase (NNMT) in pericytes and patient prognosis, as well as its association with CTC-neutrophil cluster formation, was assessed using clinical specimens and public CRC datasets. The formation process of CTC-neutrophil clusters was visualised using intravital microscope and microfluidic vascular chip models. Bulk RNA sequencing, xenograft and allograft models and pericyte-specific genetic deficiency mice were used to investigate the effect of pericyte NNMT on CTC-neutrophil cluster formation and CRC metastasis.
CTC-neutrophil clusters formed at the vascular-immune interface of CRC primary tumours. Pericytes with high NNMT expression could serve as a poor prognostic biomarker that indicated an increased risk of developing metastasis in CRC. NNMT highly NNMT-expressing pericytes facilitated the formation of CTC-neutrophil clusters by mediating the cellular interaction among CRC cells, neutrophils and endothelial cells through activating the CXCL5/CXCR2 axis. Genetic and pharmacological inhibition of NNMT in pericytes eliminated CTC-neutrophil clusters and suppressed CRC liver metastasis.
This study uncovers the previously undefined location and mechanism of CTC-neutrophil cluster formation and underscores the potential of pericyte-driven CTC-neutrophil clusters as a valuable prognostic indicator and therapeutic target for CRC metastasis.
循环肿瘤细胞（CTC）-中性粒细胞簇是肿瘤转移的关键驱动因素和标志；然而，仍缺乏有效的消除方法。
本研究旨在阐明CTC-中性粒细胞簇形成的位置和机制，并开发更有效的抗转移策略。
对临床结直肠癌（CRC）样本进行免疫荧光染色，以确定CTC-中性粒细胞簇的位置。使用临床标本和公开的CRC数据集评估周细胞中烟酰胺N-甲基转移酶（NNMT）的表达与患者预后之间的相关性，以及其与CTC-中性粒细胞簇形成的关联。使用活体显微镜和微流控血管芯片模型观察CTC-中性粒细胞簇的形成过程。采用批量RNA测序、异种移植和同种异体移植模型以及周细胞特异性基因缺陷小鼠，研究周细胞NNMT对CTC-中性粒细胞簇形成和CRC转移的影响。
CTC-中性粒细胞簇在CRC原发肿瘤的血管-免疫界面形成。高表达NNMT的周细胞可作为预后不良的生物标志物，表明CRC发生转移的风险增加。高表达NNMT的周细胞通过激活CXCL5/CXCR2轴介导CRC细胞、中性粒细胞和内皮细胞之间的细胞相互作用，促进CTC-中性粒细胞簇的形成。对周细胞中NNMT进行基因和药物抑制可消除CTC-中性粒细胞簇，并抑制CRC肝转移。
本研究揭示了CTC-中性粒细胞簇形成的先前未明确的位置和机制，并强调了周细胞驱动的CTC-中性粒细胞簇作为CRC转移的有价值的预后指标和治疗靶点的潜力。
