來自Rag2基因缺失多能干細胞的抗原特異性TCR-T細胞在小鼠模型中阻礙實體瘤的生長

2023年，中國科學院廣州生物醫學與健康研究所，中國科學院再生生物學重點實驗室，廣東省干細胞與再生醫學重點實驗室，中國科學院大學，北京，中國科學院動物研究所，干細胞與再生研究所，干細胞與生殖生物學國家重點實驗室，廣州市荔灣人民醫院，北京干細胞與再生醫學研究所，生物土地實驗室(廣州再生醫學與健康廣東實驗室)（CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicineand Health, Chinese Academy of Sciences, Guangzhou, China

University of Chinese Academy of Sciences, Beijing, ChinaState Key Laboratory of Stem Cell and Reproductive Biology, Institute for Stem Cell and Regeneration, Institute of Zoology, Chinese Academy of Sciences, Beijing,China

Liwan People's Hospital of Guangzhou, Guangzhou, China

Beijing Institute for Stem Cell and Regenerative Medicine, Beijing, China

Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou, China） Hongling Wu研究團隊在《Cell Proliferation》上，發表論文，標題為：“Antigen-specific TCR-T cells from Rag2 gene-deleted pluripotent stem cells impede solid tumour growth in a mouse model”
“來自Rag2基因缺失多能干細胞的抗原特異性TCR-T細胞在小鼠模型中阻礙實體瘤的生長”

Abstract

The technology of adoptive transfer of T‐cell receptor (TCR) engineered T cells is wildly investigated as it has the potential to treat solid cancers. However, the therapeutic application of TCR‐T cells is hampered by the poor quality derived mainly from patients' peripheral blood, as well as heterogeneous TCRs caused by the mismatch between transgenic and endogenous TCRs. To improve the homogeneity, antigen‐specificity and reduce possible autoreactivity, here we developed a technique to generate antigen‐specific T cells from Rag2 gene‐deleted pluripotent stem cells (PSCs) and further measured their anti‐tumour efficacy. PSCs were first targeted with OT1 TCR into the Rag2 locus to prevent TCR rearrangement during T‐cell development. The engineered PSCs were then differentiated through a two‐step strategy, in vitro generation of haematopoietic progenitor cells, and in vivo development and maturation of TCR‐T cells. Finally, the response to tumour cells was assessed in vitro and in vivo. The regenerated OT1‐iT displayed monoclonal antigen‐specific TCR expression, and phonotypic normalities in the spleen and lymph node tissues. Importantly, the OT1‐iT cells eliminated tumour cells while releasing specific cytokines in vitro. Furthermore, adoptive transfer of OT1‐iT cells suppresses solid tumour growth in tumour‐bearing animals. Our study presents a novel and straightforward strategy for producing antigen‐specific TCR‐T cells in vivo from PSCs, allowing for allogeneic transplantation and therapy of solid tumours.

摘要

T細胞受體(TCR)工程T細胞過繼轉移技術由于具有治療實體腫瘤的潛力而受到廣泛的研究。然而，主要來自患者外周血的TCR - T細胞質量較差，以及轉基因TCR與內源性TCR不匹配導致的TCR - T細胞異質性，阻礙了TCR - T細胞的治療應用。為了提高同質性、抗原特異性和降低可能的自身反應性，我們開發了一種從Rag2基因缺失的多能干細胞(PSCs)中生成抗原特異性T細胞的技術，并進一步測量了它們的抗腫瘤功效。首先將OT1 TCR靶向PSCs進入Rag2位點，以防止T細胞發育過程中TCR重排。然后通過兩步策略分化工程化的PSCs，體外生成造血祖細胞，體內發育和成熟TCR - T細胞。最后，在體外和體內評估對腫瘤細胞的反應。再生的OT1‐iT顯示單克隆抗原特異性TCR表達，脾臟和淋巴結組織的音型正常。重要的是，OT1‐iT細胞在體外釋放特異性細胞因子的同時消除腫瘤細胞。此外，OT1 - iT細胞的過繼性轉移抑制了荷瘤動物實體瘤的生長。我們的研究提出了一種新穎而直接的從PSCs體內產生抗原特異性TCR - T細胞的策略，允許同種異體移植和實體腫瘤的治療。

該論文中，效應T細胞與靶細胞E.G7‐OVA（小鼠T 淋巴瘤細胞系）的體外培養是使用Ausbian特級胎牛血清完成的。欲了解或購買Ausbian特級胎牛血清可以聯系北京締一生物400-166-8600.