使用整合到肝细胞癌细胞基因组中的HBV DNA表达谱来选择用于

StephenW

Gastroenterology. 2019 Jan 31. pii: S0016-5085(19)30348-8. doi: 10.1053/j.gastro.2019.01.251. [Epub ahead of print]
Use of Expression Profiles of HBV DNA Integrated Into Genomes of Hepatocellular Carcinoma Cells to Select T Cells for Immunotherapy.
Tan AT1, Yang N2, Krishnamoorthy TL3, Oei V1, Chua A4, Xinyuan Z4, Si TH4, Chia A1, Le Bert N1, Low D5, Tan HK3, Kumar R3, Irani FG6, Zong HZ4, Zhang Q7, Guccione E5, Lu-En W8, Koh S8, Hwang W9, Chow WC3, Bertoletti A10.
Author information

1
    Emerging Infectious Diseases, Duke-NUS Medical School, Singapore.
2
    Genome Institute of Singapore, Agency for Science and Technology (A*STAR), Singapore.
3
    Department of Gastroenterology & Hepatology, Singapore General Hospital, Singapore.
4
    Lion TCR Pte Ltd, Singapore.
5
    Institute of Molecular and Cell Biology, Agency for Science and Technology (A*STAR), Singapore.
6
    Department of Vascular & Interventional Radiology, Singapore General Hospital, Singapore.
7
    Department of Biotherapy, The Third Affiliated Hospital of Sun Yat-Sen University, Guandong, China.
8
    Lion TCR Pte Ltd, Singapore; Singapore Immunology Network, Agency for Science and Technology (A*STAR), Singapore.
9
    Department of Haematology, Singapore General Hospital, Singapore.
10
    Emerging Infectious Diseases, Duke-NUS Medical School, Singapore; Singapore Immunology Network, Agency for Science and Technology (A*STAR), Singapore. Electronic address: [email protected].

Abstract
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 mRNAs by real-time PCR, 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 (1x104-10x106 TCR+ T cells/kg) weekly for 112 days or 1 year. We monitored patients' liver function, serum levels of cytokines, and standard blood parameters. Anti-tumor 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.

Copyright © 2019 AGA Institute. Published by Elsevier Inc. All rights reserved.
KEYWORDS:

Adoptive T-cell transfer; HCC; TCR; TCR T-cell

PMID:
    30711630
DOI:
    10.1053/j.gastro.2019.01.251

StephenW

消化内科。 2019年1月31日.pii：S0016-5085（19）30348-8。 doi：10.1053 / j.gastro.2019.01.251。 [印刷前的电子版]
使用整合到肝细胞癌细胞基因组中的HBV DNA表达谱来选择用于免疫疗法的T细胞。
Tan AT1，Yang N2，Krishnamoorthy TL3，Oei V1，Chua A4，Xinyuan Z4，Si TH4，Chia A1，Le Bert N1，Low D5，Tan HK3，Kumar R3，Irani FG6，Zong HZ4，Zhang Q7，Guccione E5，Lu -En W8，Koh S8，Hwang W9，Chow WC3，Bertoletti A10。
作者信息

1
    新加坡杜克 - 新加坡国立大学医学院新出现的传染病。
2
    新加坡基因组研究所，新加坡科学技术局（A * STAR）。
3
    新加坡新加坡总医院消化内科和肝病学系。
4
    Lion TCR Pte Ltd，新加坡。
五
    新加坡科学技术局（A * STAR）分子与细胞生物学研究所。
6
    新加坡新加坡综合医院血管与介入放射科。
7
    中山大学附属第三医院生物治疗科，广东省。
8
    Lion TCR Pte Ltd，新加坡;新加坡免疫学网络，科学技术局（A * STAR），新加坡。
9
    新加坡新加坡总医院血液科。
10
    新加坡杜克 - 新加坡国立大学医学院新出现的传染病;新加坡免疫学网络，科学技术局（A * STAR），新加坡。电子地址：[email protected]。

抽象
背景与目的：

肝细胞癌（HCC）通常与乙型肝炎病毒（HBV）感染有关。大多数HBV相关HCC的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中的存在是否可用于选择用于免疫疗法的HBV特异性T细胞受体（TCR）。
方法：

基于免疫组织化学对HBV抗原呈阴性的HCC细胞通过实时PCR，测序和纳米串方法分析HBV 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例肝移植术后HCC复发患者的原发性HCC和转移灶的HBV基因表达谱。使用HBV转录物谱，我们从先前由具有自限性HBV感染的患者表征的TCR文库中选择对由检测到的HBV mRNA编码的HBV表位特异的TCR。通过将mRNA电穿孔进入细胞，将自体T细胞工程化以表达所选择的TCR，并且将这些TCR T细胞过量转移至患者，每周增加数量（1×10 4 -10×10 6 TCR + T细胞/ kg），持续112天或1年。我们监测患者的肝功能，血清细胞因子水平和标准血液参数。基于甲胎蛋白的血清水平和转移的计算机断层扫描评估抗肿瘤功效。
结果：

不表达完整HBV抗原的HCC细胞含有短HBV mRNA，其编码被HBV特异性T细胞识别并激活HBV特异性T细胞的表位。对患有转移的HBV DNA表达的表位特异性表达TCR的自体T细胞给予2名没有明显不良事件的患者。细胞在1年内不影响肝功能。在1名患者中，6名肺转移瘤中的5名在T细胞给药的1年期间体积减少。
结论：

HCC细胞包含整合的HBV DNA的短片段，其编码被T细胞识别并激活T细胞的表位。这些细胞的HBV转录组可用于改造T细胞用于个体化免疫疗法。该方法可用于治疗更广泛的HBV相关HCC患者。

版权所有©2019 AGA Institute。由Elsevier Inc.出版。保留所有权利。
关键词：

过继性T细胞转移; HCC; TCR; TCR T细胞

结论：
    30711630
DOI：
    10.1053 / j.gastro.2019.01.251

StephenW

https://www.gastrojournal.org/article/S0016-5085(19)30348-8/pdf