NCCRP1 缺失克服肺腺癌的免疫逃逸
Loss of NCCRP1 overcomes immune evasion in lung adenocarcinoma.
作者
作者单位
- Anhui Provincial Key Laboratory of Immunology in Chronic Diseases, Bengbu Medical University, Bengbu, Anhui, China.
- Bengbu Medical University Key Laboratory of Cancer Research and Clinical Laboratory Diagnosis, School of Laboratory Medicine, Bengbu Medical University, Bengbu, Anhui, China.
- Anhui Provincial Key Laboratory of Immunology in Chronic Diseases, Bengbu Medical University, Bengbu, Anhui, China linweiwei@bbmu.edu.cn.
摘要
中文
肺腺癌患者对免疫治疗的低反应率主要是由于肿瘤免疫逃逸。肿瘤免疫抑制微环境协调这种逃逸，但潜在机制仍不清楚。在这里，我们确定了非特异性细胞毒性细胞受体蛋白 1（NCCRP1）作为这一过程的关键且此前未表征的调节因子。我们首先分析了公开可用的患者来源的单细胞 RNA 测序和 RNA 测序数据，以研究 NCCRP1 在肺腺癌中的表达及其对肿瘤免疫微环境的影响。随后，使用 CRISPR-Cas9 在小鼠肺腺癌细胞中敲除 Nccrp1 基因。然后使用 NSG 和 C57BL/6 小鼠的皮下移植模型评估 NCCRP1 缺失对肿瘤生长的影响。采用单细胞 RNA 测序、流式细胞术和多种靶向体内干预来评估 NCCRP1 对肿瘤免疫微环境的影响。为了探索 NCCRP1 调节肿瘤免疫微环境的潜在机制，我们进行了免疫共沉淀、RNA pull-down、泛素化测定、质谱、相对和绝对定量同位素标记蛋白质组学、双荧光素酶报告基因测定和 ELISA。NCCRP1 缺失在免疫活性 C57BL/6 小鼠模型中抑制肺肿瘤生长并延长生存。NCCRP1 缺陷上调 CX3CL1 以招募 CX3CR1+ 抗肿瘤巨噬细胞。这些巨噬细胞随后分泌 CXCL9 和 CXCL10，增强 CD8+ T 细胞和 NK 细胞向肿瘤微环境的浸润。机制上，NCCRP1 和 STAU1 竞争性结合 NEDD4。NCCRP1 缺陷增强 STAU1-NEDD4 相互作用，促进 STAU1 的泛素化和蛋白酶体降解，从而增加 CX3CL1 信使 RNA 稳定性。值得注意的是，消除 NCCRP1 与抗程序性细胞死亡蛋白 1 或干扰素-γ 治疗协同作用，导致完全肿瘤根除。我们的发现强调靶向 NCCRP1 是重新编程免疫抑制微环境和克服肺腺癌“冷肿瘤”表型的一种有前景的治疗策略。
English
The low response rate to immunotherapy in patients with lung adenocarcinoma is primarily due to tumor immune evasion. The tumor immunosuppressive microenvironment orchestrates this evasion, yet the underlying mechanisms remain elusive. Here we identify non-specific cytotoxic cell receptor protein 1 (NCCRP1) as a critical and previously uncharacterized regulator of this process. We first analyzed publicly accessible patient-derived single-cell RNA sequencing and RNA sequencing data to investigate the expression of NCCRP1 in lung adenocarcinoma and its impact on the tumor immune microenvironment. Subsequently, the Nccrp1 gene was knocked out in mouse lung adenocarcinoma cells using CRISPR-Cas9. The effect of NCCRP1 deletion on tumor growth was then evaluated using subcutaneous transplantation models in both NSG and C57BL/6 mice. Single-cell RNA sequencing, flow cytometry and multiple targeted in vivo interventions were employed to assess the influence of NCCRP1 on the tumor immune microenvironment. To explore the underlying mechanisms by which NCCRP1 regulates the tumor immune microenvironment, we conducted co-immunoprecipitation, RNA pull-down, ubiquitination assay, mass spectrometry, isobaric tags for relative and absolute quantitation proteomics, dual-luciferase reporter gene assay, and ELISA. Loss of NCCRP1 inhibits lung tumor growth and prolongs survival in immunocompetent C57BL/6 mouse models. NCCRP1 deficiency upregulates CX3CL1 to recruit CX3CR1+ antitumoral macrophages. These macrophages subsequently secrete CXCL9 and CXCL10, enhancing the infiltration of CD8+ T cells and NK cells into the tumor microenvironment. Mechanistically, NCCRP1 and STAU1 competitively bind to NEDD4. NCCRP1 deficiency enhances the STAU1-NEDD4 interaction, promoting ubiquitination and proteasomal degradation of STAU1, thereby increasing CX3CL1 messenger RNA stability. Remarkably, ablating NCCRP1 synergized with anti-programmed cell death protein 1 or interferon-γ therapy, leading to complete tumor eradication. Our findings highlight NCCRP1 targeting as a promising therapeutic strategy to reprogram the immunosuppressive microenvironment and overcome the "cold tumor" phenotype in lung adenocarcinoma.
分类与指标
- 研究类型
- 基础研究
- 病种
- 肺癌
- JCR 分区
- Q1
- 影响因子
- 11.7
- 新锐分区
- 1区