肿瘤坏死因子受体 2 通过调节乳腺癌化疗耐药中的糖酵解促进内皮细胞介导的 CD8+T 细胞抑制。

Tumor necrosis factor receptor 2 promotes endothelial cell-mediated suppression of CD8+ T cells through tuning glycolysis in chemoresistance of breast cancer.

机构信息

Henan China-Germany International Joint Laboratory of Tumor Immune Microenvironment and Disease, Medical Research Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China.

Department of Medicine, Luohe Medical College, Luohe, 462000, China.

出版信息

J Transl Med. 2024 Jul 20;22(1):672. doi: 10.1186/s12967-024-05472-5.

DOI:10.1186/s12967-024-05472-5

PMID:39033271

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11265105/

Abstract

BACKGROUND

T cells play a pivotal role in chemotherapy-triggered anti-tumor effects. Emerging evidence underscores the link between impaired anti-tumor immune responses and resistance to paclitaxel therapy in triple-negative breast cancer (TNBC). Tumor-related endothelial cells (ECs) have potential immunoregulatory activity. However, how ECs regulate T cell activity during TNBC chemotherapy remains poorly understood.

METHODS

Single-cell analysis of ECs in patients with TNBC receiving paclitaxel therapy was performed using an accessible single-cell RNA sequencing (scRNA-seq) dataset to identify key EC subtypes and their immune characteristics. An integrated analysis of a tumor-bearing mouse model, immunofluorescence, and a spatial transcriptome dataset revealed the spatial relationship between ECs, especially Tumor necrosis factor receptor (TNFR) 2+ ECs, and CD8+ T cells. RNA sequencing, CD8+ T cell proliferation assays, flow cytometry, and bioinformatic analyses were performed to explore the immunosuppressive function of TNFR2 in ECs. The downstream metabolic mechanism of TNFR2 was further investigated using RNA sequencing, cellular glycolysis assays, and western blotting.

RESULTS

In this study, we identified an immunoregulatory EC subtype, characterized by enhanced TNFR2 expression in non-responders. By a mouse model of TNBC, we revealed a dynamic reduction in the proportion of the CD8+ T cell-contacting tumor vessels that could co-localize spatially with CD8+ T cells during chemotherapy and an increased expression of TNFR2 by ECs. TNFR2 suppresses glycolytic activity in ECs by activating NF-κB signaling in vitro. Tuning endothelial glycolysis enhances programmed death-ligand (PD-L) 1-dependent inhibitory capacity, thereby inducing CD8+ T cell suppression. In addition, TNFR2+ ECs showed a greater spatial affinity for exhausted CD8+ T cells than for non-exhausted CD8+ T cells. TNFR2 blockade restores impaired anti-tumor immunity in vivo, leading to the loss of PD-L1 expression by ECs and enhancement of CD8+ T cell infiltration into the tumors.

CONCLUSIONS

These findings reveal the suppression of CD8+ T cells by ECs in chemoresistance and indicate the critical role of TNFR2 in driving the immunosuppressive capacity of ECs via tuning glycolysis. Targeting endothelial TNFR2 may serve as a potent strategy for treating TNBC with paclitaxel.

摘要

背景

T 细胞在化疗引发的抗肿瘤作用中发挥关键作用。新出现的证据强调了在三阴性乳腺癌（TNBC）中，抗肿瘤免疫反应受损与紫杉醇治疗耐药之间的联系。肿瘤相关的内皮细胞（ECs）具有潜在的免疫调节活性。然而，ECs 在 TNBC 化疗过程中如何调节 T 细胞活性仍知之甚少。

方法

使用可及的单细胞 RNA 测序（scRNA-seq）数据集对接受紫杉醇治疗的 TNBC 患者的 ECs 进行单细胞分析，以鉴定关键的 EC 亚型及其免疫特征。在荷瘤小鼠模型、免疫荧光和空间转录组数据集的综合分析中，揭示了 ECs（尤其是肿瘤坏死因子受体（TNFR）2+ECs）与 CD8+T 细胞之间的空间关系。进行 RNA 测序、CD8+T 细胞增殖测定、流式细胞术和生物信息学分析，以探讨 EC 中 TNFR2 的免疫抑制功能。进一步使用 RNA 测序、细胞糖酵解测定和 Western blot 研究 TNFR2 的下游代谢机制。

结果

在这项研究中，我们鉴定了一种免疫调节型 EC 亚型，其特征是无反应者中 TNFR2 表达增强。通过 TNBC 小鼠模型，我们揭示了在化疗过程中，与 CD8+T 细胞接触的肿瘤血管的比例动态减少，并且 EC 中 TNFR2 的表达增加，这与 CD8+T 细胞在空间上共定位。TNFR2 通过激活 NF-κB 信号通路在体外抑制 EC 的糖酵解活性。调节内皮糖酵解可增强程序性死亡配体（PD-L）1 依赖性抑制能力，从而诱导 CD8+T 细胞抑制。此外，TNFR2+ECs 与耗竭的 CD8+T 细胞比非耗竭的 CD8+T 细胞具有更大的空间亲和力。TNFR2 阻断在体内恢复了受损的抗肿瘤免疫，导致 EC 中 PD-L1 表达丧失，并增强 CD8+T 细胞浸润肿瘤。