Gramantieri L, Montagner A, Arleo A, Suzzi F, Bassi C, Tovoli F, Bruccoleri M, Alimenti E, Fornari F, Iavarone M, Negrini M, Piscaglia F, Giovannini C
Division of Internal Medicine, Hepatobiliary and Immunoallergic Diseases, IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy.
Department of Medical and Surgical Sciences, Bologna University, Bologna, Italy.
ESMO Open. 2025 Mar;10(3):104289. doi: 10.1016/j.esmoop.2025.104289. Epub 2025 Mar 5.
Advanced hepatocellular carcinoma (HCC) has a dismal prognosis; however, the introduction of atezolizumab-bevacizumab combination has improved overall survival and novel immune checkpoint inhibitors are entering the clinics. Despite more therapeutic options being available, no biomarker guides treatment choice. Indeed, tissue-based analyses and complex analytical procedures hinder clinical translation. We explored the informativeness of a simple, non-invasive, repeatable cytofluorimetric assay on peripheral blood to predict response and survival in HCC patients treated with atezolizumab-bevacizumab.
Twenty-five cirrhotic patients, 50 HCC patients undergoing atezolizumab-bevacizumab and an independent validation cohort of 25 HCC patients were subjected to a cytofluorimetric study of peripheral white blood cells (WBCs) to assess baseline programmed death-ligand 1-positive (PD-L1+) and cytotoxic T-lymphocyte antigen 4-positive (CTLA4+) cell percentage in the different populations and their early on-treatment variations. Immunophenotypes were evaluated against treatment response. RNA sequencing followed by RT-PCR validation were used to elucidate the molecular correlates of immunophenotypic observations.
PD-L1+ cell percentage did not predict response either at baseline or when evaluating treatment-induced early changes. Conversely, the percentage of CTLA4+ lymphocytes at baseline showed a predictive significance (35.37 in responders versus 31.5 in non-responders, P = 0.03). More interestingly, the early CTLA4+ changes during treatment in lymphocytes (responders 0.95 versus non-responders 1.08, P = 0.05), monocytes (responders 0.95 versus non-responders 1.04, P = 0.03), granulocytes (responders 0.94 versus non-responders 1.14, P = 0.001) and, even stronger, the early CTLA4+ percentage change in the whole WBCs displayed a predictive significance in terms of time to progression (TTP) (P < 0.0001) and overall survival (OS) (P = 0.005). The immunophenotypic findings correlated with transcriptional modulation of CTLA4 target genes and genes involved in immune response.
A repeatable, easy, non-invasive blood test predicts response to immunotherapy in patients with HCC, both in terms of TTP and OS. CTLA4+ cell percentage increase in non-responders suggests a possible resistance mechanism which deserves attention as a druggable target.
晚期肝细胞癌（HCC）预后不佳；然而，阿替利珠单抗-贝伐珠单抗联合疗法的引入改善了总生存期，并且新型免疫检查点抑制剂正在进入临床。尽管有更多的治疗选择，但尚无生物标志物可指导治疗选择。实际上，基于组织的分析和复杂的分析程序阻碍了临床转化。我们探索了一种简单、无创、可重复的外周血细胞荧光分析方法在预测接受阿替利珠单抗-贝伐珠单抗治疗的HCC患者的反应和生存情况方面的信息价值。
对25例肝硬化患者、50例接受阿替利珠单抗-贝伐珠单抗治疗的HCC患者以及一个由25例HCC患者组成的独立验证队列进行外周血白细胞（WBC）的细胞荧光分析，以评估不同群体中基线程序性死亡配体1阳性（PD-L1+）和细胞毒性T淋巴细胞抗原4阳性（CTLA4+）细胞的百分比及其治疗早期的变化。针对治疗反应评估免疫表型。采用RNA测序并通过RT-PCR验证来阐明免疫表型观察结果的分子相关性。
无论是在基线时还是评估治疗引起的早期变化时，PD-L1+细胞百分比均不能预测反应。相反，基线时CTLA4+淋巴细胞的百分比显示出预测意义（反应者为35.37，无反应者为31.5，P = 0.03）。更有趣的是，治疗期间淋巴细胞（反应者为0.95，无反应者为1.08，P = 0.05）、单核细胞（反应者为0.95，无反应者为1.04，P = 0.03）、粒细胞（反应者为0.94，无反应者为1.14，P = 0.001）中CTLA4+的早期变化，甚至更明显的是，全白细胞中CTLA4+百分比的早期变化在疾病进展时间（TTP）（P < 0.0001）和总生存期（OS）（P = 0.005）方面显示出预测意义。免疫表型结果与CTLA4靶基因和参与免疫反应的基因的转录调控相关。
一种可重复、简便、无创的血液检测可预测HCC患者对免疫治疗的反应，包括TTP和OS。无反应者中CTLA4+细胞百分比增加提示一种可能的耐药机制，作为一个可药物作用靶点值得关注。
