PO.CL01.05 · 临床研究
cadonilimab联合regorafenib作为晚期肝细胞癌二线或后线治疗的最新探索性生物标志物分析:确定Bola3为潜在预测性生物标志物
Updated exploratory biomarker analysis of cadonilimab plus regorafenib as second-line or later therapy in advanced hepatocellular carcinoma: Identification of Bola3 as a potential predictive biomarker
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景:cadonilimab是一种同类首创的靶向PD-1和CTLA-4的双特异性抗体,与regorafenib(一种已获批用于肝细胞癌(HCC)二线治疗的多激酶抑制剂)联合使用,在经过大量既往治疗的晚期HCC(aHCC)患者中显示出良好的抗肿瘤活性和可控的安全性。我们此前在2025年AACR年会上报告了最新的临床结局(摘要#CT172,NCT05644379)。在此,我们介绍一项探索性生物标志物分析的最新发现,重点关注BOLA3在治疗反应中的潜在预测作用。
方法:在治疗前和治疗期间采集配对血浆样本,进行游离RNA测序(cfRNA-seq)以评估转录动态。使用DESeq2进行差异表达分析(FDR ≤ 0.10),并应用Hallmark GSEA评估线粒体和免疫相关通路的变化。ELISA在基线和治疗早期定量检测可溶性PD-1和CTLA-4,计算个体数值。在预先设定的患者亚组中,对基线肿瘤组织进行半定量BOLA3免疫组织化学(IHC)。对BOLA3、TOMM20、PD-L1、CD8alpha和DAPI进行多重免疫荧光(mIF),实现细胞水平的定量以及肿瘤与瘤周空间分布的比较。
结果:ELISA检测到治疗前和治疗中可溶性PD-1和CTLA-4水平稳定,并观察到早期个体动态;然而,这些水平与治疗反应无相关性。cfRNA-seq显示,治疗后PR患者的BOLA3降低,而PD患者的BOLA3升高。通路分析表明,PD相关的线粒体信号富集且炎症反应较弱,而PR则呈现相反趋势。基线BOLA3 IHC与转录组学发现一致:疾病进展患者的BOLA3表达高于疾病控制患者,作为连续变量分析时仍保持方向性关联。初步mIF提示,反应者表现出较低的BOLA3/TOMM20和较高的PD-L1/CD8,而进展者则表现出相反的表型。
结论:综合探索性分析将BOLA3提名为候选的结局相关生物标志物,并支持这样一个模型:反应者表现出线粒体低、炎症高的表型。这些发现提供了早期的、互补的证据,表明BOLA3相关特征可能有助于患者分层,进一步的验证正在进行中。研究资助方:天津市自然科学基金联合基金(编号25JCLZJC00120)。
查看英文原文 English abstract
Background: Cadonilimab, a first-in-class bispecific antibody targeting PD-1 and CTLA-4, in combination with regorafenib, a multikinase inhibitor approved for second-line treatment of hepatocellular carcinoma (HCC), has demonstrated promising antitumor activity and manageable safety in heavily pretreated patients with advanced HCC (aHCC). We previously reported the updated clinical outcomes at the 2025 AACR Annual Meeting (Abstract #CT172, NCT05644379). Here, we present updated findings from an exploratory biomarker analysis focusing on the potential predictive role of BOLA3 in treatment response.
Methods: Paired plasma samples were collected before and during treatment for cell-free RNA sequencing (cfRNA-seq) to evaluate transcriptional dynamics. Differential expression analysis was performed using DESeq2 (FDR ≤ 0.10), and Hallmark GSEA was applied to assess mitochondrial and immune-related pathway changes. ELISA quantified soluble PD-1 and CTLA-4 at baseline and early on-treatment, calculating individual values. In a prespecified patient subset, baseline tumor tissues underwent semi-quantitative BOLA3 immunohistochemistry (IHC). Multiplex immunofluorescence (mIF) was performed for BOLA3, TOMM20, PD-L1, CD8alpha, and DAPI, enabling cell-level quantification and comparison of tumor versus peritumoral spatial distribution.
Results: ELISA detected stable levels of soluble PD-1 and CTLA-4 pre- and on-treatment, with early individual dynamics observed; however, these levels did not correlate with treatment response. cfRNA-seq revealed decreased BOLA3 in the PR patient and increased BOLA3 in the PD patient after treatment. Pathway analysis indicated PD-associated enrichment of mitochondrial signaling and weaker inflammatory response, whereas PR showed the opposite trend. Baseline BOLA3 IHC aligned with transcriptomic findings: higher BOLA3 expression was observed in patients with disease progression versus disease control, maintaining directional association when analyzed as a continuous variable. Preliminary mIF suggested that responders exhibited lower BOLA3/TOMM20 and higher PD-L1/CD8, while progressors showed the reverse phenotype.
Conclusions: Integrated exploratory analyses nominate BOLA3 as a candidate outcome-associated biomarker and support a model in which responders exhibit a mitochondria-low, inflamed-high phenotype. These findings provide early, complementary evidence that BOLA3-related features may inform patient stratification, with further validation underway. Research Sponsor: The Joint Funds of the Natural Science Foundation of Tianjin (No.25JCLZJC00120).
利益披露 Disclosure
Y. Liu, None..
J. Zhang, None..
Q. Qi, None..
D. Wu, None..
L. Xu, None..
Y. Li, None..
H. Li, None.