Hepatocellular carcinoma (HCC) is a formidable disease, distinguished by its high aggressiveness and dismal outcomes. Although leucine aminopeptidase (LAP) has been widely employed as a biomarker in biological imaging of HCC, it is still susceptible to interference from false-positive signals activated in injured liver tissues. In this study, based on the significant difference of GSH levels in alcohol-damaged liver tissues and tumor tissues, a dual-tandem activatable probe (PCLT) was designed for differential diagnosis and treatment guidance of HCC by near-infrared fluorescence (NIRF) imaging. This probe comprised a dual-locked hemicyanine dye decorated with a tetraethylene glycol chain and dual-recognition unit of glutathione (GSH) and LAP, which could be sequentially cleaved by GSH and LAP to restore its NIRF signal. PCLT excellently discriminated orthotopic HCC from ALI far earlier (7 days) than histological analysis (28 days) and exhibited higher specificity toward early orthotopic HCC than the single-locked probe (PCL). In addition, PCLT is capable of accurately delineating the tumor contour, assisting in surgical resection of HCC tumors under fluorescence visualization, and noninvasively assessing the antitumor effect of HCC chemotherapy during ferroptosis, thereby presenting promising clinical implications for clinical diagnosis and therapy of HCC.

中文翻译：

---

用于肝细胞癌鉴别诊断和治疗评估的串联可激活荧光探针的合理设计


肝细胞癌 （HCC） 是一种可怕的疾病，以其高侵袭性和令人沮丧的结果而著称。尽管亮氨酸氨肽酶 （LAP） 已被广泛用作 HCC 生物成像中的生物标志物，但它仍然容易受到受伤肝组织中激活的假阳性信号的干扰。本研究基于酒精损伤肝组织和肿瘤组织中 GSH 水平的显著差异，设计了双串联激活探针 （PCLT），用于近红外荧光 （NIRF） 成像对 HCC 的鉴别诊断和治疗指导。该探针由装饰有四乙二醇链的双锁半菁染料和谷胱甘肽 （GSH） 和 LAP 的双重识别单元组成，可以被 GSH 和 LAP 依次切割以恢复其 NIRF 信号。PCLT 出色地区分原位 HCC 与 ALI 的时间远早于组织学分析 （7 天） （28 天），并且对早期原位 HCC 的特异性高于单锁探针 （PCL）。此外，PCLT 能够准确描绘肿瘤轮廓，在荧光可视化下辅助手术切除 HCC 肿瘤，并在铁死亡过程中无创评估 HCC 化疗的抗肿瘤效果，从而为 HCC 的临床诊断和治疗带来有希望的临床意义。