Department of Surgery, Experimental Surgery, CCM, CVK, Charité - Universitätsmedizin Berlin, Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany.
Department of Pathology, CCM, Charité - Universitätsmedizin Berlin, Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany.
Oncoimmunology. 2024 Oct 1;13(1):2406052. doi: 10.1080/2162402X.2024.2406052. eCollection 2024.
Intrahepatic cholangiocarcinoma (ICC) is a disease with poor prognosis and limited therapeutic options. We investigated the tumor immune microenvironment (TIME) to identify predictors of disease outcome and to explore targets for therapeutic modulation.
Liver tissue samples were collected during 2008-2019 from patients ( = 139) diagnosed with ICC who underwent curative intent surgery without neoadjuvant chemotherapy. Samples from the discovery cohort ( = 86) were immunohistochemically analyzed on tissue microarrays (TMAs) for the expression of CD68, CD3, CD4, CD8, Foxp3, PD-L1, STAT1, and p-STAT1 in tumor core and stroma areas. Results were digitally analyzed using QuPath software and correlated with clinicopathological characteristics. For validation of TIME-related biomarkers, we performed multiplex imaging mass cytometry (IMC) in a validation cohort ( = 53).
CD68+ cells were the predominant immune cell type in the TIME of ICC. CD4+ T cell density correlated with better overall survival (OS). Prediction modeling together with validation cohort confirmed relevance of CD4+ cells, PD-L1 expression by immune cells in the stroma and N-stage on overall disease outcome. In turn, IMC analyses revealed that silent CD3+CD4+ clusters inversely impacted survival. Among annotated immune cell clusters, PD-L1 was most relevantly expressed by CD4+FoxP3+ cells. A subset of tumors with high density of immune cells ("hot" cluster) correlated with PD-L1 expression and could identify a group of candidates for immune checkpoint inhibition (ICI). Ultimately, higher levels of STAT1 expression were associated with higher lymphocyte infiltration and PD-L1 expression.
These results highlight the importance of CD4+ T cells in immune response against ICC. Secondly, a subset of tumors with "hot" TIME represents potential candidates for ICI, while stimulation of STAT1 pathway could be a potential target to turn "cold" into "hot" TIME in ICC.
肝内胆管癌（ICC）是一种预后不良且治疗选择有限的疾病。我们研究了肿瘤免疫微环境（TIME），以确定疾病结局的预测因子，并探索治疗调节的靶点。
2008 年至 2019 年间，从接受无新辅助化疗的根治性手术治疗的 ICC 患者（n=139）中收集肝组织样本。在发现队列（n=86）中，使用组织微阵列（TMA）对 CD68、CD3、CD4、CD8、Foxp3、PD-L1、STAT1 和 p-STAT1 在肿瘤核心和基质区域的表达进行免疫组织化学分析。使用 QuPath 软件对结果进行数字分析，并与临床病理特征相关联。为了验证与 TIME 相关的生物标志物，我们在验证队列（n=53）中进行了多重成像质谱细胞术（IMC）分析。
CD68+细胞是 ICC TIME 中的主要免疫细胞类型。CD4+T 细胞密度与总生存（OS）相关。预测模型与验证队列共同证实了 CD4+细胞、基质中免疫细胞的 PD-L1 表达以及 N 期对整体疾病结局的相关性。反过来，IMC 分析显示，沉默的 CD3+CD4+簇对生存产生负面影响。在注释的免疫细胞簇中，PD-L1 最相关地表达于 CD4+Foxp3+细胞。具有高密度免疫细胞的肿瘤亚组（“热”簇）与 PD-L1 表达相关，可鉴定出一组接受免疫检查点抑制（ICI）的候选者。最终，更高水平的 STAT1 表达与更高的淋巴细胞浸润和 PD-L1 表达相关。
这些结果强调了 CD4+T 细胞在针对 ICC 的免疫反应中的重要性。其次，具有“热”TIME 的肿瘤亚组代表了 ICI 的潜在候选者，而激活 STAT1 通路可能是将 ICC 的“冷”TIME 转化为“热”TIME 的潜在靶点。
