通過誘導型半胱天冬酶 9 消耗 T 細胞提高了過繼 T 細胞療法

StephenW

通過誘導型半胱天冬酶 9 消耗 T 細胞提高了過繼 T 細胞療法對抗慢性乙型肝炎的安全性
亞歷山大·克洛普 1 2 3 ，索菲亞·施賴伯 1 2 ，安娜 D 科辛斯卡 1 2 3 ，馬丁·普萊 4 ，烏爾里克·普羅策 1 2 3 ，卡琳·維斯基辛 1 2 3
隸屬關係
隸屬關係

    1
    德國慕尼黑工業大學病毒學研究所醫學院。
    2
    德國慕尼黑 Helmholtz Zentrum München 病毒學研究所。
    3
    德國感染研究中心 (DZIF)，慕尼黑合作夥伴站點，德國慕尼黑。
    4
    英國倫敦大學學院癌症研究所血液學系。

    PMID：34691041 PMCID：PMC8527178 DOI：10.3389/fimmu.2021.734246

抽象的

使用通過病毒特異性受體重定向到乙型肝炎病毒 (HBV) 感染細胞的 T 細胞進行 T 細胞療法是治療慢性乙型肝炎和 HBV 相關癌症的一種有前途的治療方法。然而，由於靶細胞數量眾多，細胞因子釋放綜合徵或肝毒性等副作用可能會限制安全性。因此，允許按需消耗轉移的 T 細胞的保護機制將是增加對這種方法的信心的有趣手段。在這項研究中，T 細胞是通過逆轉錄病毒轉導產生的，以表達 HBV 特異性嵌合抗原受體 (S-CAR) 或 T 細胞受體 (TCR)，此外還有誘導型半胱天冬酶 9 (iC9) 或單純皰疹病毒胸苷激酶 (HSV-TK) 作為安全開關。使用複制 HBV 的肝癌細胞作為靶點的實時細胞毒性分析表明，兩個安全開關的激活都在不到一小時內停止了 S-CAR 或 TCR 轉導的 T 細胞的細胞毒性。在體內，iC9 的誘導導致轉移的 S-CAR T 細胞過繼轉移到 AAV-HBV 感染的免疫無能小鼠中的強烈和快速減少。注射 iC9 二聚體後 1 到 6 小時，觀察到血液和脾臟中的 S-CAR T 細胞減少了 90% 以上，肝臟中的 S-CAR T 細胞減少了 99% 以上，從而限制了肝毒性並阻止了細胞因子的分泌。然而，同時，S-CAR T 細胞的抗病毒作用減弱，因為剩餘的 S-CAR T 細胞大多無功能，不能用 HBsAg 重新刺激。 iC9 的第二次誘導只能消耗肝臟中的 T 細胞。總之，共表達 iC9 和 HBV 特異性受體的 T 細胞可有效識別和殺死 HBV 複製細胞。通過 iC9 誘導 T 細胞死亡被證明是一種在體外和體內消耗轉移 T 細胞的有效手段，其中含有不需要的肝毒性。

關鍵詞：T細胞治療；嵌合抗原受體（CAR）；慢性乙型肝炎;誘導型半胱天冬酶 9；保護分子；自殺開關。

版權所有 © 2021 Klopp、Schreiber、Kosinska、Pulé、Protzer 和 Wisskirchen。
利益衝突聲明

KW 和 UP 是 SCG Cell Therapy Pte. Ltd. 的聯合創始人和股東。 Ltd. KW 部分受僱於 SCG Cell Therapy GmbH。其餘作者聲明，該研究是在沒有任何可能被解釋為潛在利益衝突的商業或財務關係的情況下進行的。

StephenW

Depletion of T cells via Inducible Caspase 9 Increases Safety of Adoptive T-Cell Therapy Against Chronic Hepatitis B
Alexandre Klopp  1   2   3 , Sophia Schreiber  1   2 , Anna D Kosinska  1   2   3 , Martin Pulé  4 , Ulrike Protzer  1   2   3 , Karin Wisskirchen  1   2   3
Affiliations
Affiliations

    1
    School of Medicine, Institute of Virology, Technical University of Munich, Munich, Germany.
    2
    Institute of Virology, Helmholtz Zentrum München, Munich, Germany.
    3
    German Center for Infection Research (DZIF), Munich Partner Site, Munich, Germany.
    4
    Department of Haematology, Cancer Institute, University College London, London, United Kingdom.

    PMID: 34691041 PMCID: PMC8527178 DOI: 10.3389/fimmu.2021.734246

Abstract

T-cell therapy with T cells that are re-directed to hepatitis B virus (HBV)-infected cells by virus-specific receptors is a promising therapeutic approach for treatment of chronic hepatitis B and HBV-associated cancer. Due to the high number of target cells, however, side effects such as cytokine release syndrome or hepatotoxicity may limit safety. A safeguard mechanism, which allows depletion of transferred T cells on demand, would thus be an interesting means to increase confidence in this approach. In this study, T cells were generated by retroviral transduction to express either an HBV-specific chimeric antigen receptor (S-CAR) or T-cell receptor (TCR), and in addition either inducible caspase 9 (iC9) or herpes simplex virus thymidine kinase (HSV-TK) as a safety switch. Real-time cytotoxicity assays using HBV-replicating hepatoma cells as targets revealed that activation of both safety switches stopped cytotoxicity of S-CAR- or TCR-transduced T cells within less than one hour. In vivo, induction of iC9 led to a strong and rapid reduction of transferred S-CAR T cells adoptively transferred into AAV-HBV-infected immune incompetent mice. One to six hours after injection of the iC9 dimerizer, over 90% reduction of S-CAR T cells in the blood and the spleen and of over 99% in the liver was observed, thereby limiting hepatotoxicity and stopping cytokine secretion. Simultaneously, however, the antiviral effect of S-CAR T cells was diminished because remaining S-CAR T cells were mostly non-functional and could not be restimulated with HBsAg. A second induction of iC9 was only able to deplete T cells in the liver. In conclusion, T cells co-expressing iC9 and HBV-specific receptors efficiently recognize and kill HBV-replicating cells. Induction of T-cell death via iC9 proved to be an efficient means to deplete transferred T cells in vitro and in vivo containing unwanted hepatotoxicity.

Keywords: T-cell therapy; chimeric antigen receptor (CAR); chronic hepatitis B; inducible caspase 9; safeguard molecules; suicide switch.

Copyright © 2021 Klopp, Schreiber, Kosinska, Pulé, Protzer and Wisskirchen.
Conflict of interest statement

KW and UP are co-founders and shareholders of SCG Cell Therapy Pte. Ltd. KW is partially employed by SCG Cell Therapy GmbH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

StephenW

https://pubmed.ncbi.nlm.nih.gov/34691041/