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《中华肺部疾病杂志》[ISSN:1006-6977/CN:61-1281/TN]

卷:

期数:

2024年06期

页码:

901-906

栏目:

论著

出版日期:

2024-12-25

文章信息/Info

Title:

Anti-tumor activity and mechanism of the novel OX40 antibody agonist in humanized mouse NSCLC model

作者:

刘 岩¹; 赵沛妍¹; 田 琳¹; 崔贺然²; 狄 娜²; 李 慧¹; 程 颖¹; 3

130012 长春,吉林省肿瘤医院肿瘤转化医学实验室¹
130012 长春,吉林省肿瘤医院生物样本库²
130012 长春,吉林省肿瘤医院胸部肿瘤内科³

Author(s):

Liu Yan¹;  Zhao Peiyan¹;  Tian Lin¹;  Cui Heran²;  Di Na²;  Li Hui¹;  Cheng Ying¹; 3.

¹Medical Oncology Translational research lab, Jilin Cancer Hospital, Changchun 130012, China; ² Biobank, Jilin Cancer Hospital, Changchun 130012, China; ³Department of Medical Thoracic Oncology, Jilin Cancer Hospital, Changchun 130012, China

关键词:

非小细胞肺癌;  人源化小鼠;  肿瘤坏死因子受体超家族4;  抗体激动剂;  转录组测序

Keywords:

Non-small cell lung cancer;  Humanized mouse;  Tumor necrosis factor receptor superfamily member 4;  ES102;  Transcriptome sequencing

分类号:

R734.2

DOI:

10.3877/cma.j.issn.1674-6902.2024.06.009

摘要:

目的 分析肿瘤坏死因子受体超家族4(tumor necrosis factor receptor superfamily member 4)(OX40)抗体激动剂(ES102)对小细胞肺癌(non-small cell lung cancer, NSCLC)人源化OX40小鼠模型的抑制作用和机制。方法 使用流式细胞术检测小鼠Lewis肺癌细胞(Lewis lung cancer cells, LLC)中CD44表达; 分别构建OX40人源化小鼠NSCLC荷瘤模型,溶媒对照组、不同剂量ES102组和结肠癌荷瘤模型,阳性溶媒对照组和阳性对照组,判断每组中ES102的抑瘤水平; 采用ELISA检测血清中干扰素-γ(interferon-γ, IFN-γ)和白介素-17(interleukin-17, IL-17); 蛋白质印迹法(Western blot)检测PI3K、NFAT、CD44、NapsinA和SYN蛋白表达; RNA转录组测序检测免疫浸润细胞表达。结果 LLC细胞中CD44阳性率99.58%,NapsinA蛋白低表达,提示LLC鼠源肺癌细胞株具有NSCLC特性。人源化OX40小鼠NSCLC模型中,ES102对NSCLC抑瘤能力无显著差异,MC38阳性对照组中注射ES102的第11天、第14天、第18天和第21天,ES102抑瘤能力与阳性溶媒对照组相比增强(P=0.0004,t=5.712; P<0.0001,t=9.368; P<0.0001,t=10.64; P<0.0001,t=13.37)。人源化小鼠血浆IL-17的含量在不同样本中表达差异显著(F=7.703,P=0.0004); 阳性对照组中IL-17分泌含量(58.24±18.11)pg/ml分别高于ES102(2 mg/ml)组(21.87±11.74)pg/ml,ES102(5 mg/ml)组(18.49±10.17)pg/ml和ES102(10 mg/ml)组(20.47±10.52)pg/ml(P<0.05); NSCLC ES102(10 mg/ml)组中IFN-γ含量高于对照组、ES102(2 mg/ml)组和ES102(5 mg/ml)组(P<0.05)。PI3K在阳性对照组中未见表达,在未处理组和ES102实验组(2 μg/kg～10 μg/kg)中高表达(P<0.05)。NFAT作为活化型T细胞的核因子,表达趋势与PI3K一致,组间差异无统计学意义(P>0.05)。转录组测序检测NSCLC和阳性对照组小鼠肿瘤组织,转录组测序和反卷积分析显示,Tregs细胞浸润比例为(1.95±0.02)%,低于阳性对照组中Tregs细胞浸润比例(7.2±0.03)%(P<0.05)。结论 ES102对NSCLC人源化小鼠模型疗效低于结肠癌,疗效差异与微环境中IL-17分泌降低及调节性T细胞浸润有关。

Abstract:

Objective To explore the efficacy and mechanism of OX40 antibody agonist(ES102)in humanized OX40 mouse NSCLC model. Methods Mouse Lewis lung cancer cells were detected using flow cytometry(Lewis lung cancer cells, CD44 expression in LLC); OX40 humanized mouse NSCLC tumor-bearing model(vehicle control group, ES102 treatment group at different doses)and colon cancer tumor-bearing model(positive vehicle control group and positive control group)were constructed respectively, To evaluate the tumor suppressive level of ES102 in different groups; Serum interferon- γ was measured by ELISA(interferon- γ, IFN- γ)and interleukin-17(interleukin-17, IL-17)secretory level; Expression of PI3K, NFAT, CD44, NapsinA and SYN by western blot(Western blot); The expression of the immune-infiltrating cells was determined by RNA transcriptome sequencing. Results The CD44 positive rate in LLC cells showed 99.58% and low expression of NapsinA protein, suggesting that LLC murine-derived lung cancer cell lines had NSCLC properties. In the humanized OX40 mouse NSCLC model, ES102 inhibited NSCLC, but on Day 11, Day 14, Day 18, and Day 21 in the positive vehicle control(P=0.0004, t=5.712; P<0.0001, t=9.368; P<0.0001, t=10.364; P<0.0001, t=13.37). The content of plasma IL-17 in humanized mice was significantly differently expressed between different samples(F=7.703, P=0.0004); The secretion content of IL-17 in the positive control group(58.24±18.11)pg/ml than the ES102(2 mg/ml)group(21.87±11.74)pg/ml, The ES102(5 mg/ml)group(18.49±10.17)pg/ml and the ES102(10 mg/ml)group(20.47±10.52)pg/ml(P<0.05); Higher IFN- γ in NSCLC ES102(10 mg/ml)than the control, ES102(2 mg/ml)and ES102(5 mg/ml)(P<0.05). PI3K expression was not seen in the positive control group and was highly expressed in the untreated group and the ES102 experimental group(2 μg/kg to 10 μg/kg)(P<0.05). As a nuclear factor in activated T cells, NFAT was expressed in a trend consistent with PI3K, without a statistically significant difference between the groups(P>0.05). After transcriptome sequencing of NSCLC and positive control mice, further transcriptome sequencing and deconvolution analysis showed that the proportion of Tregs cells infiltrated was(1.95±0.02)%, which was lower than that of Tregs cells infiltrated in the positive control group(7.2±0.03)%(P<0.05). Conclusion The efficacy of ES102 in humanized NSCLC mice is lower than colon cancer, and the difference may be related to the decreased secretion of IL-17 and the infiltration of Tregs cells in the microenvironment.

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备注/Memo

备注/Memo:

基金项目: 吉林省科技厅基础研究专项(202002063JC)
吉林省卫健委项目(2021JC094)
通信作者: 李 慧, Email: 181966963@qq.com
程 颖, Email: chengying@csco.org.cn

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更新日期/Last Update: 2024-12-25