Pediatric Surgical Oncology Laboratory, Michael E. DeBakey Department of Surgery, Divisions of Pediatric Surgery and Surgical Research, Texas Children's Surgical Oncology Program, Texas Children's Liver Tumor Program, Dan L. Duncan Cancer Center, Baylor College of Medicine, Houston, Texas, USA.
Department of Pathology and Immunology, Baylor College of Medicine, Texas Children's Department of Pathology, Houston, Texas, USA.
Hepatol Commun. 2024 May 10;8(6). doi: 10.1097/HC9.0000000000000435. eCollection 2024 Jun 1.
Hepatoblastoma and HCC are the most common malignant hepatocellular tumors seen in children. The aim of this study was to develop a liquid biopsy test for circulating tumor cells (CTCs) for these tumors that would be less invasive and provide real-time information about tumor response to therapy.
For this test, we utilized indocyanine green (ICG), a far-red fluorescent dye used clinically to identify malignant liver cells during surgery. We assessed ICG accumulation in cell lines using fluorescence microscopy and flow cytometry. For our CTC test, we developed a panel of liver tumor-specific markers, including ICG, Glypican-3, and DAPI, and tested it with cell lines and noncancer control blood samples. We then used this panel to analyze whole-blood samples for CTC burden with a cohort of 15 patients with hepatoblastoma and HCC and correlated with patient characteristics and outcomes.
We showed that ICG accumulation is specific to liver cancer cells, compared to nonmalignant liver cells, non-liver solid tumor cells, and other nonmalignant cells, and can be used to identify liver tumor cells in a mixed population of cells. Experiments with the ICG/Glypican-3/DAPI panel showed that it specifically tagged malignant liver cells. Using patient samples, we found that CTC burden from sequential blood samples from the same patients mirrored the patients' responses to therapy.
Our novel ICG-based liquid biopsy test for CTCs can be used to specifically detect and quantify CTCs in the blood of pediatric patients with liver cancer.
肝母细胞瘤和 HCC 是儿童中最常见的恶性肝细胞肿瘤。本研究旨在为这些肿瘤开发一种循环肿瘤细胞（CTC）的液体活检测试，这种测试方法创伤性更小，并能实时提供肿瘤对治疗反应的信息。
对于该测试，我们利用了吲哚菁绿（ICG），这是一种远红色荧光染料，临床上用于在手术期间识别恶性肝细胞。我们使用荧光显微镜和流式细胞术评估了 ICG 在细胞系中的积累情况。对于我们的 CTC 测试，我们开发了一组肝脏肿瘤特异性标志物，包括 ICG、Glypican-3 和 DAPI，并在细胞系和非癌症对照血液样本中对其进行了测试。然后，我们使用该面板分析了 15 名肝母细胞瘤和 HCC 患者的全血样本中的 CTC 负担，并与患者特征和结果相关联。
与非恶性肝细胞、非肝脏实体瘤细胞和其他非恶性细胞相比，我们发现 ICG 积累是肝癌细胞特有的，并且可以用于在混合细胞群中识别肝肿瘤细胞。使用 ICG/Glypican-3/DAPI 面板的实验表明，它可以特异性标记恶性肝细胞。使用患者样本，我们发现来自同一患者的连续血液样本中的 CTC 负担反映了患者对治疗的反应。
我们新的基于 ICG 的 CTC 液体活检测试可用于特异性检测和定量儿童癌症患者血液中的 CTC。
