High-throughput Sequencing Quantitative Analysis of altered genes expression in bladder cancer with anti-PD-1 treatment

Immune-checkpoint blockade (ICB) therapy has profoundly changed the landscape of cancer treatment strategies. Despite its success, resistance remains a significant challenge, with the majority of patients exhibiting non-response to ICB therapies. ICB reinvigorates tumor-specific T cells by interrupting inhibitory signals mediated by checkpoints like PD-1 and CTLA-4, pivotal for the reactivation of their anti-tumor functions. Within the tumor microenvironment, tumor-specific T cells enter a state of dysfunction, making it difficult to generate an effective response to ICB. This study aims to investigate the altered transcriptome in anti-PD-1 responders and non-responders of bladder cancer (BCa) and to uncover the underlying mechanisms of T cell non-responses to ICB. Overall design: To explore T cell dysfunction contributing to ICB resistance in BCa, we developed an immunocompetent spontaneous BCa mouse model. After tumor formation, anti-PD-1 monoclonal antibody was applied to treat BCa. Total RNA from responsive and non-responsive BCa was extracted, and high-throughput sequencing was conducted to investigate the expression profile differences between non-responders and responders.

免疫检查点阻断（Immune-checkpoint blockade, ICB）疗法深刻重塑了癌症治疗策略的整体格局。尽管该疗法已取得显著临床成效，但耐药性仍是亟待攻克的重大难题——多数患者对ICB治疗无应答。ICB通过阻断PD-1、CTLA-4等免疫检查点介导的抑制信号，重振肿瘤特异性T细胞的抗肿瘤活性，而这些免疫检查点正是调控T细胞抗肿瘤功能再激活的关键靶点。在肿瘤微环境中，肿瘤特异性T细胞会进入功能耗竭状态，使得机体难以对ICB治疗产生有效应答。本研究旨在分析膀胱癌（bladder cancer, BCa）抗PD-1治疗应答者与无应答者的差异转录组，并揭示T细胞对ICB治疗无应答的潜在分子机制。实验设计方案：为探究膀胱癌中介导ICB耐药的T细胞功能耗竭机制，我们构建了免疫健全的自发性膀胱癌小鼠模型。待肿瘤形成后，采用抗PD-1单克隆抗体对模型小鼠进行治疗。分别提取抗PD-1治疗应答组与无应答组小鼠的膀胱癌组织总RNA，通过高通量测序分析两组间的基因表达谱差异。