ING4肿瘤抑制因子的缺陷与晚期肿瘤和癌症患者生存率低有关。显示ING4在几种癌症中抑制NF-kB。由于NF-kB是免疫应答的关键介质,ING4/NF-kB轴可能在肿瘤免疫调节中表现,但尚未进行研究.为了表征与ING4缺陷肿瘤相关的肿瘤免疫微环境,本研究采用了三种方法:第一,由246个原发性乳腺肿瘤(包括97个ING4缺陷型肿瘤)组成的组织微阵列评估了选择性免疫标志物的存在,CD68,CD4,CD8和PD-1,使用免疫组织化学染色。第二,利用CRISPR介导的来自Tp53缺失的乳腺肿瘤细胞系中Ing4的缺失,设计了一种免疫活性小鼠ING4缺陷型乳腺癌模型;我们使用流式细胞术评估了乳腺肿瘤的免疫标志物.最后,对METABRIC基因表达数据集评估了与Ing4缺失肿瘤相关的免疫标志物相关的患者生存率.结果显示CD68、CD4、CD8或PD-1与ING4缺陷型乳腺肿瘤无显著相关性,表明巨噬细胞没有富集,T细胞,或耗尽的T细胞类型。在老鼠身上,Ing4缺失的乳腺肿瘤的生长速率与Ing4完整的肿瘤相当,但显示出增加的肿瘤渗透率和转移。Ing4缺失肿瘤的免疫标记分析显示肿瘤相关巨噬细胞(Gr-1loCD11bF4/80)显着增加,颗粒酶B阳性(GzmB)CD4T细胞减少,表明抑制和/或较少的肿瘤杀伤免疫微环境。METABRIC数据分析显示GZMB的低表达与患者的低生存率显著相关。ING4低表达,在乳腺癌的基底亚型中。GZMB低/ING4低肿瘤患者的生存结局最差(HR=2.80,95%CI1.36-5.75,p=0.0004),支持GZMB低免疫环境有助于ING4缺陷肿瘤进展的观点。总的来说,研究结果表明,ING4缺陷型肿瘤存在有助于免疫逃避和转移的微环境.
Deficiencies in the ING4 tumor suppressor are associated with advanced stage tumors and poor patient survival in cancer. ING4 was shown to inhibit NF-kB in several cancers. As NF-kB is a key mediator of immune response, the ING4/NF-kB axis is likely to manifest in tumor-immune modulation but has not been investigated. To characterize the tumor immune microenvironment associated with ING4-deficient tumors, three approaches were employed in this study: First, tissue microarrays composed of 246 primary breast tumors including 97 ING4-deficient tumors were evaluated for the presence of selective immune markers, CD68, CD4, CD8, and PD-1, using immunohistochemical staining. Second, an immune-competent mouse model of ING4-deficient breast cancer was devised utilizing CRISPR-mediated deletion of Ing4 in a Tp53 deletion-derived mammary tumor cell line; mammary tumors were evaluated for immune markers using flow cytometry. Lastly, the METABRIC gene expression dataset was evaluated for patient survival related to the immune markers associated with Ing4-deleted tumors. The results showed that CD68, CD4, CD8, or PD-1, was not significantly associated with ING4-deficient breast tumors, indicating no enrichment of macrophages, T cells, or exhausted T cell types. In mice, Ing4-deleted mammary tumors had a growth rate comparable to Ing4-intact tumors but showed increased tumor penetrance and metastasis. Immune marker analyses of Ing4-deleted tumors revealed a significant increase in tumor-associated macrophages (Gr-1loCD11b+F4/80+) and a decrease in granzyme B-positive (GzmB+) CD4+ T cells, indicating a suppressive and/or less tumoricidal immune microenvironment. The METABRIC data analyses showed that low expression of GZMB was significantly associated with poor patient survival, as was ING4-low expression, in the basal subtype of breast cancer. Patients with GZMB-low/ING4-low tumors had the worst survival outcomes (HR = 2.80, 95% CI 1.36-5.75, p = 0.0004), supportive of the idea that the GZMB-low immune environment contributes to ING4-deficient tumor progression. Collectively, the study results demonstrate that ING4-deficient tumors harbor a microenvironment that contributes to immune evasion and metastasis.