肝内胆管细胞癌免疫微环境的临床预后指标与靶点

Clinical prognosticators and targets in the immune microenvironment of intrahepatic cholangiocarcinoma

OncoImmunology

影响因子:6.3

分区:医学2区 / 免疫学2区肿瘤学2区

发表日期:2024

作者: Isis Lozzi, Alexander Arnold, Matthias Barone, Juliette Claire Johnson, Bruno V Sinn, Johannes Eschrich, Pimrapat Gebert, Ruonan Wang, Mengwen Hu, Linda Feldbrügge, Anja Schirmeier, Anja Reutzel-Selke, Thomas Malinka, Felix Krenzien, Wenzel Schöning, Dominik P Modest, Johann Pratschke, Igor M Sauer, Matthäus Felsenstein

DOI: 10.1080/2162402X.2024.2406052

摘要

肝内胆管细胞癌（ICC）是一种预后较差、治疗选择有限的疾病。我们研究了肿瘤免疫微环境（TIME），旨在寻找疾病预后的预测因子及治疗调控的靶点。收集了2008年至2019年期间诊断为ICC、接受根治性手术且未接受新辅助化疗的患者（n=139）的肝组织样本。通过免疫组化分析，使用组织微阵列（TMAs）检测肿瘤核心区和基质区的CD68、CD3、CD4、CD8、Foxp3、PD-L1、STAT1和p-STAT1的表达。结果采用QuPath软件进行数字分析，并与临床病理特征相关联。为了验证TIME相关生物标志物，在验证队列（n=53）中进行多重成像质谱（IMC）分析。结果显示，CD68+细胞是ICC TIME中最主要的免疫细胞类型。CD4+高密度T细胞与更好的总生存期（OS）相关。预测模型结合验证队列确认了CD4+细胞、基质中免疫细胞的PD-L1表达以及N期对总体预后的重要性。IMC分析表明，无声的CD3+CD4+簇对生存具有负面影响。在所有免疫细胞簇中，PD-L1最主要由CD4+FoxP3+细胞表达。具有高免疫细胞密度（“热”簇）的肿瘤与PD-L1表达相关，可能成为免疫检查点抑制（ICI）的候选对象。最终，较高的STAT1表达水平与淋巴细胞浸润和PD-L1表达增加相关。这些结果强调了CD4+ T细胞在抗ICC免疫反应中的重要作用。其次，具有“热”免疫微环境的肿瘤是潜在的ICI候选，激活STAT1通路可能成为将“冷”转变为“热”微环境的潜在靶点。

Abstract

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 (n = 139) diagnosed with ICC who underwent curative intent surgery without neoadjuvant chemotherapy. Samples from the discovery cohort (n = 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 (n = 53).CD68+ cells were the predominant immune cell type in the TIME of ICC. CD4+high 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.