Xu Yulin, Shan Wei, Luo Qian, Zhang Meng, Huo Dawei, Chen Yijin, Li Honghu, Ye Yishan, Yu Xiaohong, Luo Yi, Huang He
Bone Marrow Transplantation Center, The First Affiliated Hospital Zhejiang University School of Medicine Hangzhou China.
Liangzhu Laboratory Zhejiang University Hangzhou China.
Cancer Innov. 2024 Apr 15;3(3):e118. doi: 10.1002/cai2.118. eCollection 2024 Jun.
Cancer-targeted T-cell receptor T (TCR-T) cells hold promise in treating cancers such as hematological malignancies and breast cancers. However, approaches to obtain cancer-reactive TCR-T cells have been unsuccessful.
Here, we developed a novel strategy to screen for cancer-targeted TCR-T cells using a special humanized mouse model with person-specific immune fingerprints. Rare steady-state circulating hematopoietic stem and progenitor cells were expanded via three-dimensional culture of steady-state peripheral blood mononuclear cells, and then the expanded cells were applied to establish humanized mice. The human immune system was evaluated according to the kinetics of dendritic cells, monocytes, T-cell subsets, and cytokines. To fully stimulate the immune response and to obtain B-cell precursor NAML-6- and triple-negative breast cancer MDA-MB-231-targeted TCR-T cells, we used the inactivated cells above to treat humanized mice twice a day every 7 days. Then, human T cells were processed for TCR β-chain (TRB) sequencing analysis. After the repertoires had been constructed, features such as the fraction, diversity, and immune signature were investigated.
The results demonstrated an increase in diversity and clonality of T cells after treatment. The preferential usage and features of TRBV, TRBJ, and the V-J combination were also changed. The stress also induced highly clonal expansion. Tumor burden and survival analysis demonstrated that stress induction could significantly inhibit the growth of subsequently transfused live tumor cells and prolong the survival of the humanized mice.
We constructed a personalized humanized mouse model to screen cancer-targeted TCR-T pools. Our platform provides an effective source of cancer-targeted TCR-T cells and allows for the design of patient-specific engineered T cells. It therefore has the potential to greatly benefit cancer treatment.
靶向癌症的T细胞受体T（TCR-T）细胞在治疗血液系统恶性肿瘤和乳腺癌等癌症方面具有前景。然而，获取癌症反应性TCR-T细胞的方法一直未成功。
在此，我们开发了一种新策略，使用具有个体特异性免疫指纹的特殊人源化小鼠模型来筛选靶向癌症的TCR-T细胞。通过稳态外周血单个核细胞的三维培养来扩增罕见的稳态循环造血干细胞和祖细胞，然后将扩增后的细胞用于建立人源化小鼠。根据树突状细胞、单核细胞、T细胞亚群和细胞因子的动力学来评估人类免疫系统。为了充分刺激免疫反应并获得靶向B细胞前体NAML-6和三阴性乳腺癌MDA-MB-231的TCR-T细胞，我们使用上述灭活细胞每7天对人源化小鼠每天治疗两次。然后，对人T细胞进行TCRβ链（TRB）测序分析。构建库后，研究诸如比例、多样性和免疫特征等特征。
结果表明治疗后T细胞的多样性和克隆性增加。TRBV、TRBJ以及V-J组合的优先使用和特征也发生了变化。应激还诱导了高度克隆性扩增。肿瘤负荷和生存分析表明，应激诱导可显著抑制随后输注的活肿瘤细胞的生长，并延长人源化小鼠的生存期。
我们构建了一个个性化的人源化小鼠模型来筛选靶向癌症的TCR-T库。我们的平台提供了靶向癌症的TCR-T细胞的有效来源，并允许设计患者特异性工程化T细胞。因此，它有可能极大地有益于癌症治疗。
