背景:据报道CD93促进肿瘤血管生成。然而,CD93是否调节抗肿瘤免疫仍不清楚。方法:肺肿瘤组织,恶性胸腔积液(MPEs)来自肺癌患者.从健康志愿者和接受抗PD-1治疗的肺癌患者获得血液。此外,p53fl/flLSL-KrasG12D,Ccr7-/-,产生Cd93-/-小鼠和CD11c-DTR小鼠。具体来说,EM,利用NTA和蛋白质印迹来鉴定肿瘤细胞外囊泡(TEV)。EV标签,体外和体内EV摄取的检测,进行EV蛋白和RNA的降解以检测TEV在肿瘤进展中的作用。分离胸膜间皮细胞(pMCs)以研究相关的信号通路。产生重组蛋白和抗体以测试哪种抗体是最有效的抗体以增加p-pMC中的CCL21a。RNA-Seq,miRNA阵列,荧光素酶报告分析,内皮管形成试验,蛋白质标记和检测,siRNA和miRNA模拟物和抑制剂的转染,趋化性测定,免疫组织化学染色,流式细胞术,实时PCR,进行ELISA实验。结果:我们显示pMC的CD93通过阻止pMC分泌CCL21来减少树突状细胞的肺肿瘤迁移,从而抑制系统性抗肺肿瘤T细胞反应。TEV衍生的miR-5110通过下调pMCCD93促进CCL21分泌,而C1q,在肿瘤个体中增加,抑制CD93介导的CCL21分泌。CD93阻断抗体(抗CD93)比VEGF受体阻断抗体更好地抑制肺肿瘤生长,因为抗CD93抑制肿瘤血管生成并促进CCL21从pMC分泌。抗CD93还克服了肺肿瘤对抗PD-1疗法的抗性。此外,血清EV来源的miR-5193(人miR-5110同源物)较高的肺癌患者对抗PD-1治疗更敏感,而血清C1q较高的患者不太敏感,与它们对CD93的调节功能一致。结论:我们的研究确定了CD93在控制抗肺肿瘤免疫中的关键作用,并为肺肿瘤治疗提供了有希望的方法。
Background: CD93 reportedly facilitates tumor angiogenesis. However, whether CD93 regulates antitumor immunity remains undeciphered. Methods: Lung tumor tissues, malignant pleural effusions (MPEs) were obtained from lung cancer patients. Blood was obtained from healthy volunteers and lung cancer patients with anti-PD-1 therapy. Furthermore, p53fl/flLSL-KrasG12D, Ccr7-/-, Cd93-/- mice and CD11c-DTR mice were generated. Specifically, EM, NTA and western blotting were utilized to identify Tumor extracellular vesicles (TEVs). EV labeling, detection of EV uptake in vitro and in vivo, degradation of EV proteins and RNAs were performed to detect the role of TEVs in tumor progression. Pleural mesothelial cells (pMCs) were isolated to investigate related signaling pathways. Recombinant proteins and antibodies were generated to test which antibody was the most effective one to increase CCL21a in p-pMCs. RNA-Seq, MiRNA array, luciferase reporter assay, endothelial tube formation assay, protein labeling and detection, transfection of siRNAs and the miRNA mimic and inhibitor, chemotaxis assay, immunohistochemical staining, flow cytometry, Real-time PCR, and ELISA experiments were performed. Results: We show that CD93 of pMCs reduced lung tumor migration of dendritic cells by preventing pMCs from secreting CCL21, thereby suppressing systemic anti-lung tumor T-cell responses. TEV-derived miR-5110 promotes CCL21 secretion by downregulating pMC CD93, whereas C1q, increasing in tumor individuals, suppresses CD93-mediated CCL21 secretion. CD93-blocking antibodies (anti-CD93) inhibit lung tumor growth better than VEGF receptor-blocking antibodies because anti-CD93 inhibit tumor angiogenesis and promote CCL21 secretion from pMCs. Anti-CD93 also overcome lung tumor resistance to anti-PD-1 therapy. Furthermore, lung cancer patients with higher serum EV-derived miR-5193 (human miR-5110 homolog) are more sensitive to anti-PD-1 therapy, while patients with higher serum C1q are less sensitive, consistent with their regulatory functions on CD93. Conclusions: Our study identifies a crucial role of CD93 in controlling anti-lung tumor immunity and suggests a promising approach for lung tumor therapy.