親和力增強的TCR工程T細胞對HBV相關肝細胞癌的體內治療作用

StephenW

In vivo therapeutic effects of affinity-improved-TCR engineered T-cells on HBV-related hepatocellular carcinoma
Qi Liu  1   2 , Ye Tian  1 , Yanyan Li  1 , Wei Zhang  3 , Wenxuan Cai  1 , Yaju Liu  3 , Yuefei Ren  1   4 , Zhaoduan Liang  1 , Peipei Zhou  1   5 , Yajing Zhang  1   2 , Yifeng Bao  1 , Yi Li  6   2
Affiliations
Affiliations

    1
    State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health, Guangzhou, China.
    2
    University of the Chinese Academy of Sciences, Beijing, China.
    3
    Hefei Institute of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Hefei, Anhui, China.
    4
    School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, China.
    5
    Institute of Physical Science and Information Technology, Anhui University, Hefei, Anhui, China.
    6
    State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health, Guangzhou, China [email protected].

    PMID: 33323464 DOI: 10.1136/jitc-2020-001748

Abstract

Background: In patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC), virus-specific cytotoxic T lymphocytes (CTLs) fail to eliminate HCC cells expressing HBV antigens. As the expression of viral antigen in HBV-associated HCC may decrease to allow tumor to escape immune attacks, we hypothesized that an HBV surface antigen (HBsAg)-specific affinity-improved-T-cell receptor (TCR) will enable T cells to target HCC more effectively than corresponding wild-type-TCR. We also postulated that TCR promiscuity can be exploited to efficiently capture HBV variants that can hinder CTL-based therapeutics.

Methods: We applied flexi-panning to isolate affinity-improved TCRs binding to a variant antigen, the human leukocyte antigen (HLA)-A*02:01-restricted nonapeptide HBs371-379-ILSPFLPLL, from libraries constructed with a TCR cloned using the decapeptide HBs370-379-SIVSPFIPLL. The potency and safety of the affinity-improved-TCR engineered T-cells (Ai-TCR-T) were verified with potentially cross-reactive human and HBV-variant peptides, tumor and normal cells, and xenograft mouse models.

Results: Ai-TCR-T cells retained cognate HBV antigen specificity and recognized a wide range of HBV genotypic variants with improved sensitivity and cytotoxicity. Cell infusions produced complete elimination of HCC without recurrence in the xenograft mouse models. Elevated accumulation of CD8+ Ai-TCR-T cells in tumors correlated with tumor shrinkage.

Conclusion: The in vitro and in vivo studies demonstrated that HBsAg-specific Ai-TCR-T cells had safety profiles similar to those of their wild-type counterparts and significantly enhanced potency. This study presents an approach to develop new therapeutic strategies for HBV-related HCC.

Keywords: CD8-Positive T-lymphocytes; adoptive; cell engineering; immunohistochemistry; immunotherapy; liver neoplasms.

© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

StephenW

親和力增強的TCR工程T細胞對HBV相關肝細胞癌的體內治療作用
劉啟1 2，葉天1，李艷艷1，張偉3，蔡文軒1，劉亞菊3，岳飛飛1 4，趙端亮1，周佩佩1 5，張亞靜1 2，一峰寶1，伊力6 2
隸屬關係
隸屬關係

    1個
    廣州市生物醫學與衛生研究所呼吸疾病國家重點實驗室，廣州。
    2
    中國科學院大學，北京。
    3
    廣州市生物醫學與健康研究所合肥市干細胞與再生醫學研究所，安徽合肥。
    4
    中國科學技術大學生命科學學院，安徽合肥
    5
    安徽大學物理科學與信息技術研究所，安徽合肥
    6
    廣州市生物醫學與健康研究所呼吸疾病國家重點實驗室，廣州[email protected]。

    PMID：33323464 DOI：10.1136 / jitc-2020-001748

抽象

背景：在患有乙型肝炎病毒（HBV）相關的肝細胞癌（HCC）的患者中，病毒特異性細胞毒性T淋巴細胞（CTL）無法消除表達HBV抗原的HCC細胞。由於乙肝病毒相關肝癌中病毒抗原的表達可能降低，從而使腫瘤能夠逃避免疫攻擊，因此我們假設乙肝病毒表面抗原（HBsAg）特異性親和力增強的T細胞受體（TCR）可使T細胞靶向HCC比相應的野生型TCR更有效。我們還假設可以利用TCR濫交來有效捕獲可能阻礙基於CTL的療法的HBV變異。

方法：我們採用彈性淘選技術從使用TCR克隆的TCR構建的文庫中分離與親和力提高的TCR結合的變異體抗原，該變體抗原是人白細胞抗原（HLA）-A * 02：01限制性非肽HBs371-379-ILSPFLPLL。十肽HBs370-379-SIVSPFIPLL。用潛在的交叉反應的人和HBV變異肽，腫瘤細胞和正常細胞以及異種移植小鼠模型驗證了親和力改良的TCR工程T細胞（Ai-TCR-T）的效力和安全性。

結果：Ai-TCR-T細胞保留了相關的HBV抗原特異性，並識別了範圍廣泛的HBV基因型變體，具有更高的敏感性和細胞毒性。在異種移植小鼠模型中，細胞輸注可完全消除HCC，而不會復發。 CD8 + Ai-TCR-T細胞在腫瘤中積累的升高與腫瘤縮小有關。

結論：體外和體內研究表明，HBsAg特異的Ai-TCR-T細胞具有與野生型相似的安全性，並且效力顯著提高。這項研究提出了一種開發針對HBV相關HCC的新治療策略的方法。

關鍵詞：CD8陽性T淋巴細胞；收養細胞工程；免疫組化;免疫療法肝腫瘤。

©作者（或其雇主）2020。CCBY-NC允許重複使用。禁止商業重複使用。查看權限。由BMJ發布。