Use of Expression Profiles of HBV-DNA Integrated Into Genomes of Hepatocellular Carcinoma Cells to Select T Cells for Immunotherapy
Use of Expression Profiles of HBV-DNA Integrated Into Genomes of Hepatocellular Carcinoma Cells to Select T Cells for Immunotherapy
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DOI：
10.1053/j.gastro.2019.01.251
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发表时间：
2019-05-01
期刊：
影响因子：
29.4
通讯作者：
Bertoletti, Antonio
中科院分区：
文献类型：
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作者：
Tan, Anthony Tanoto;Yang, Ninghan;Bertoletti, Antonio
背景与目的：肝细胞癌（HCC）常与乙型肝炎病毒（HBV）感染相关。大多数HBV相关的hcc细胞含有不编码整个HBV抗原的HBV- dna片段。我们研究了这些整合的HBV-DNA片段是否编码被T细胞识别的表位，以及它们在hcc中的存在是否可以用于选择hbv特异性T细胞受体（tcr）进行免疫治疗。方法：基于免疫组织化学，通过实时聚合酶链反应、测序和纳米链方法分析HBV抗原阴性的HCC细胞中HBV信使rna （mrna）的存在。我们使用HBV特异性T细胞和tcr样抗体测试了HBV mrna阳性HCC细胞产生T细胞识别的表位的能力。然后，我们分析了2例肝移植后肝癌复发的原发性HCC和转移灶的HBV基因表达谱。利用HBV转录谱，我们从先前从自限性HBV感染患者中鉴定的TCR库中选择了检测到的HBV mRNA编码的HBV表位特异性的TCR。通过将mRNA电穿孔入细胞，对自体T细胞进行工程化，以表达所选的TCR，并将这些TCR T细胞过继性地转移到患者体内，每周增加1 × 10(4)-10 × 10(6)个TCR+ T细胞/kg，持续112天或1年。我们监测了患者的肝功能、血清细胞因子水平和标准血液参数。根据血清甲胎蛋白水平和转移灶的计算机断层扫描来评估抗肿瘤疗效。结果：不表达全HBV抗原的HCC细胞含有短的HBV mrna，其编码的表位可被HBV特异性T细胞识别并激活。对2例患者进行自体T细胞工程改造，使其表达针对转移患者HBV-DNA表位表达的特异性tcr，无明显不良事件。这些细胞在1年的时间内没有影响肝功能。在1例患者中，6个肺转移灶中有5个在给予t细胞治疗的1年期间体积减小。结论：HCC细胞含有整合的HBV-DNA短片段，其编码的表位可被T细胞识别并激活。这些细胞的HBV转录组可用于设计用于个性化免疫治疗的T细胞。这种方法可能用于治疗更广泛的hbv相关HCC患者。
BACKGROUND & AIMS: Hepatocellular carcinoma (HCC) is often associated with hepatitis B virus (HBV) infection. Cells of most HBV-related HCCs contain HBV-DNA fragments that do not encode entire HBV antigens. We investigated whether these integrated HBV-DNA fragments encode epitopes that are recognized by T cells and whether their presence in HCCs can be used to select HBV-specific T-cell receptors (TCRs) for immunotherapy. METHODS: HCC cells negative for HBV antigens, based on immunohistochemistry, were analyzed for the presence of HBV messenger RNAs (mRNAs) by real-time polymerase chain reaction, sequencing, and Nanostring approaches. We tested the ability of HBV mRNA-positive HCC cells to generate epitopes that are recognized by T cells using HBV-specific T cells and TCR-like antibodies. We then analyzed HBV gene expression profiles of primary HCCs and metastases from 2 patients with HCC recurrence after liver transplantation. Using the HBV-transcript profiles, we selected, from a library of TCRs previously characterized from patients with self-limited HBV infection, the TCR specific for the HBV epitope encoded by the detected HBV mRNA. Autologous T cells were engineered to express the selected TCRs, through electroporation of mRNA into cells, and these TCR T cells were adoptively transferred to the patients in increasing numbers (1 x 10(4)-10 x 10(6) TCR+ T cells/kg) weekly for 112 days or 1 year. We monitored patients' liver function, serum levels of cytokines, and standard blood parameters. Antitumor efficacy was assessed based on serum levels of alpha fetoprotein and computed tomography of metastases. RESULTS: HCC cells that did not express whole HBV antigens contained short HBV mRNAs, which encode epitopes that are recognized by and activate HBV-specific T cells. Autologous T cells engineered to express TCRs specific for epitopes expressed from HBV-DNA in patients' metastases were given to 2 patients without notable adverse events. The cells did not affect liver function over a 1-year period. In 1 patient, 5 of 6 pulmonary metastases decreased in volume during the 1-year period of T-cell administration. CONCLUSIONS: HCC cells contain short segments of integrated HBV-DNA that encodes epitopes that are recognized by and activate T cells. HBV transcriptomes of these cells could be used to engineer T cells for personalized immunotherapy. This approach might be used to treat a wider population of patients with HBV-associated HCC.