Wang Jiaqi, Zhu Jiying, Hu Junchi, Wang Ziruoyu, Wang Xiaobo, Pan Jianbo, Chu Yiwei, Li Zengxia, Jiang Wei, Liang Chunmin, Hou Jun, Guo Jianming, Dang Yongjun, Jiang Shuai
Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education and Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China.
Department of Immunology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China.
Genes Dis. 2022 Nov 26;10(6):2586-2596. doi: 10.1016/j.gendis.2022.10.022. eCollection 2023 Nov.
Bladder cancer (BLCA) remains a difficult malignancy to manage because of its high recurrence, intense follow-up, and invasive diagnostic and treatment techniques. Immune checkpoint inhibitors (ICIs) have forged a new direction for the treatment of BLCA, but it is currently challenging to predict whether an individual patient will be sensitive to ICIs. We collected 43 urine/tumor samples from BLCA patients for primary bladder cancer cells (BCCs) culturing using our previously reported BCC culture platform. We used flow cytometry (FCM) to measure the expression levels of Programmed Death-Ligand 1 (PD-L1) on BCCs before and after interferon-gamma (IFN-γ) treatment and found that PD-L1 expression and the sensitivities to IFN-γ varied among patients. RNA-sequencing, western blotting, and programmed death-1 (PD-1) binding assays confirmed that the BCC FCM-based PD-L1 detection platform (BC-PD-L1) was reliable and was not hindered by the glycosylation of PD-L1. In the subsequent retrospective study, we found that IFN-γ-stimulated PD-L1 (sPD-L1) expression on BCCs detected by BC-PD-L1 could predict the prognosis of BLCA patients. Importantly, the prognostic value was similar or even better in urine-derived BC-PD-L1 (UBC-PD-L1). Transcriptome analysis showed that BCCs with high sPD-L1 tended to enrich genes associated with the collagen-containing extracellular matrix, cell-cell adhesion, and positive regulation of the immune system. In addition, the UBC-PD-L1 also exhibited predictive value for ICI response in BLCA patients. In conclusion, as a novel personalized urine-detection method, UBC-PD-L1 may provide a rapid, accurate, and non-invasive tool for monitoring tumor progression, predicting therapeutic responses, and helping improve BLCA clinical treatment in future.
膀胱癌（BLCA）由于其高复发率、密集的随访以及侵入性的诊断和治疗技术，仍然是一种难以管理的恶性肿瘤。免疫检查点抑制剂（ICIs）为BLCA的治疗开辟了新方向，但目前预测个体患者是否对ICIs敏感具有挑战性。我们使用之前报道的膀胱癌细胞（BCCs）培养平台，从BLCA患者中收集了43份尿液/肿瘤样本用于原代膀胱癌细胞培养。我们使用流式细胞术（FCM）测量干扰素-γ（IFN-γ）处理前后BCCs上程序性死亡配体1（PD-L1）的表达水平，发现PD-L1表达以及对IFN-γ的敏感性在患者之间存在差异。RNA测序、蛋白质印迹和程序性死亡-1（PD-1）结合试验证实，基于BCC FCM的PD-L1检测平台（BC-PD-L1）是可靠的，并且不受PD-L1糖基化的影响。在随后的回顾性研究中，我们发现通过BC-PD-L1检测到的BCCs上IFN-γ刺激的PD-L1（sPD-L1）表达可以预测BLCA患者的预后。重要的是，尿液来源的BC-PD-L1（UBC-PD-L1）的预后价值相似甚至更好。转录组分析表明，具有高sPD-L1的BCCs倾向于富集与含胶原蛋白的细胞外基质、细胞间粘附和免疫系统正调控相关的基因。此外，UBC-PD-L1对BLCA患者的ICI反应也具有预测价值。总之，作为一种新型的个性化尿液检测方法，UBC-PD-L1可能为监测肿瘤进展、预测治疗反应以及在未来帮助改善BLCA临床治疗提供一种快速、准确且无创的工具。
