{
  "abstract": "Background Hepatocellular carcinoma (HCC) remains a leading cause of cancer mortality worldwide. While immune checkpoint blockade has revolutionized the treatment of many cancers, responses in HCC remain limited. Robust functional platforms capable of predicting individual responses to immunotherapy are urgently needed. In this study, we developed a patient-derived organotypic tumor spheroid (PDOTS) model that preserves the heterogeneity and immune microenvironment of HCC, enabling rapid and reliable assessment of targeted and immunotherapeutic responses.Methods Tumor tissues from 30 HCC patients were processed using a “Five-Point Clock” sampling method and cultured within a three-dimensional microfluidic chip supplemented with IL-2 and CD3/28 activator to maintain tumor-infiltrating lymphocyte activity. The genomic, immune, and histopathological fidelity of PDOTS relative to parental tumors was evaluated. Drug responses were assessed ex vivo and validated in matched patient-derived xenograft (PDX) models. Transcriptomic profiling was subsequently performed to identify gene signatures associated with treatment sensitivity and to construct a transcriptomic predictive model.Results The PDOTS retained ≥60% viability over 7 days and faithfully maintained the genomic, immune, and histopathological profiles of parental tumors. Functionally, PDOTS exhibited immune-dependent responses to PD-1 blockade and predicted treatment responses with 80% concordance in matched PDX models. Transcriptomic profiling revealed distinct metabolic and immune signatures in sensitive and resistant tumors, which were used to derive the “Organoid Killing Index (OKI)”. The OKI gene-derived index, a composite index integrating the enrichment scores of gene signatures associated with the OKI, strongly correlated ( R=0.829, p<0.001) and predicted clinical outcomes in an external patient cohort treated with atezolizumab plus bevacizumab.Conclusions These findings establish PDOTS as an immune-competent ex vivo platform for functional precision oncology and support the integration of functional testing with transcriptomic prediction to guide individualized immunotherapy in HCC.",
  "authors": [
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China",
        "Department of Hepatobiliary Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China"
      ],
      "name": "Fei Song"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China"
      ],
      "name": "Yi-Jun Lu"
    },
    {
      "affiliations": [
        "Liver Cancer Institute, Zhongshan Hospital, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China"
      ],
      "name": "Hai-Xiang Sun"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China"
      ],
      "name": "Jian-Wen Cheng"
    },
    {
      "affiliations": [
        "Shanghai-MOST Key Laboratory of Health and Disease Genomics, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Fudan University, Shanghai, China"
      ],
      "name": "Wen-Zhen Li"
    },
    {
      "affiliations": [
        "Shanghai-MOST Key Laboratory of Health and Disease Genomics, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Fudan University, Shanghai, China"
      ],
      "name": "Zhenhao Liu"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China"
      ],
      "name": "Rui-Zhe Li"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China"
      ],
      "name": "Peng-Xiang Wang"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China"
      ],
      "name": "Wen-Jing Zheng"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China"
      ],
      "name": "Yu-Chen Zhong"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China"
      ],
      "name": "Yang Xu"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China"
      ],
      "name": "Jie Xu"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China"
      ],
      "name": "Yu Wang"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China"
      ],
      "name": "Su-Yan Bai"
    },
    {
      "affiliations": [
        "Shanghai-MOST Key Laboratory of Health and Disease Genomics, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Fudan University, Shanghai, China"
      ],
      "name": "Lu Xie"
    },
    {
      "affiliations": [
        "Department of Surgery, School of Clinical Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China"
      ],
      "name": "Kwan Man"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China",
        "Institutes of Biomedical Sciences, Fudan University, Shanghai, China"
      ],
      "name": "Jia Fan"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China",
        "Institutes of Biomedical Sciences, Fudan University, Shanghai, China"
      ],
      "name": "Jian Zhou"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China"
      ],
      "name": "Zhong Chen"
    },
    {
      "affiliations": [
        "Department of Hepatobiliary Surgery & Liver Transplantation, Liver Cancer Institute, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital Fudan University, Shanghai, China"
      ],
      "name": "Xin-Rong Yang"
    }
  ],
  "title": "Patient-derived organotypic tumor spheroids as a functional platform for predicting immunotherapeutic responses and guiding precision treatment for hepatocellular carcinoma",
  "uid": "f9f93b14-e491-59aa-b094-13f65455e1d2"
}