Shang Bingqing, Guo Liping, Shen Rongfang, Cao Chuanzhen, Xie Ruiyang, Jiang Weixing, Wen Li, Bi Xingang, Shi Hongzhe, Zheng Shan, Li Changling, Ma Jianhui, Zhang Kaitai, Feng Lin, Shou Jianzhong
Department of Urology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Peking University People's Hospital, Peking University Institute of Hematology, National Clinical Research Center for Hematologic Disease, Beijing Key Laboratory of Hematopoietic Stem Cell Transplantation, Beijing, China.
Front Oncol. 2021 Dec 21;11:771545. doi: 10.3389/fonc.2021.771545. eCollection 2021.
Non-metastatic renal cell carcinoma (RCC) with tumor thrombus showed a greater tendency for developing metastases after surgery. Early identification of patients with high risk of poor prognosis is especially important to explore adjuvant treatment of improving outcomes. Neutrophil-to-lymphocyte ratio (NLR) was a systemic inflammation marker and outcome predictor in RCC, reflecting the chaos in systemic immune status in cancer as myeloid cell expansion and lymphatic cell suppression. Neutrophil extracellular traps (NET) formation (NETosis) is the process of neutrophils generating an extracellular DNA net-like structure. NETosis in tumor was demonstrated to conduce to the subsequent metastases of tumor. However, the role of NLR for systemic immune status and tumor local immune infiltration, especially for neutrophil-associated NETs, in non-metastatic RCC with thrombus remains unclear.
In our clinical cohort, we enrolled the clinical, pathologic, and preoperative laboratory parameters of 214 RCC patients with tumor thrombus who were treated surgically. The clinical endpoint was defined as cancer-specific survival (CSS). In our basic research cohort, RNA-seq, TCR-seq, and scRNA-seq data were analyzed. Patients who reached the endpoint as recurrence-free survival (RFS) were defined as the "High-risk" group. Otherwise, they were separated into the "Low-risk" group.
In the clinical cohort, NLR≥4 was an independent risk factor for 203 localized RCC with tumor thrombus. In the basic research cohort, tumor thrombi were separated into NETosis-thrombi belonging to the "High-risk" group and non-NETosis-thrombi to the "Low-risk" group. NETs induced by tumor-derived G-CSF in tumor thrombus has a mechanistic role in unfavorable prognosis. Besides, NETs-score from single sample GSEA (ssGSEA) algorithm was an independent prognostic factor validated in the TCGA data. Apart from the neutrophils-associated NETosis, systemic immune perturbations of lymphocytes occurred in the "High-risk" group, represented with decreased TCR diversity and increasingly high proportion of CD4-positive effector memory T (Tem) cells, which indirectly represented the state of lymphopenia.
Our findings firstly demonstrated that neutrophils-associated NETosis and systemic lymphocytes perturbations were considered as tumor progression in patients of localized RCC with tumor thrombus, which reflected NLR≥4 as an independent risk factor for patients.
伴有肿瘤血栓的非转移性肾细胞癌（RCC）术后发生转移的倾向更大。早期识别预后不良的高危患者对于探索改善预后的辅助治疗尤为重要。中性粒细胞与淋巴细胞比值（NLR）是RCC中的一种全身炎症标志物和预后预测指标，反映了癌症中全身免疫状态的紊乱，即髓样细胞扩张和淋巴细胞抑制。中性粒细胞胞外陷阱（NET）形成（NETosis）是中性粒细胞产生细胞外DNA网状结构的过程。肿瘤中的NETosis被证明有助于肿瘤的后续转移。然而，NLR在伴有血栓的非转移性RCC中对全身免疫状态和肿瘤局部免疫浸润，尤其是对中性粒细胞相关NETs的作用仍不清楚。
在我们的临床队列中，我们纳入了214例接受手术治疗的伴有肿瘤血栓的RCC患者的临床、病理和术前实验室参数。临床终点定义为癌症特异性生存（CSS）。在我们的基础研究队列中，分析了RNA测序、TCR测序和单细胞RNA测序数据。达到无复发生存（RFS）终点的患者被定义为“高危”组。否则，他们被分为“低危”组。
在临床队列中，NLR≥4是203例伴有肿瘤血栓的局限性RCC的独立危险因素。在基础研究队列中，肿瘤血栓被分为属于“高危”组的NETosis血栓和属于“低危”组的非NETosis血栓。肿瘤血栓中肿瘤来源的粒细胞集落刺激因子诱导的NETs在不良预后中起作用。此外，单样本基因集富集分析（ssGSEA）算法得出的NETs评分是在TCGA数据中得到验证的独立预后因素。除了中性粒细胞相关的NETosis外，“高危”组还出现了淋巴细胞的全身免疫紊乱，表现为TCR多样性降低和CD4阳性效应记忆T（Tem）细胞比例越来越高，这间接代表了淋巴细胞减少的状态。
我们的研究结果首次表明，中性粒细胞相关的NETosis和全身淋巴细胞紊乱被认为是伴有肿瘤血栓的局限性RCC患者的肿瘤进展，这表明NLR≥4是患者的独立危险因素。
