利用基因工程病人来源的淋巴细胞表达肿瘤反应性受体进行免疫治疗是一种有前景的恶性肿瘤治疗方法。然而，在这种治疗中使用自体T细胞限制了基因工程T细胞的质量，从而限制了细胞的及时输注给病人。本研究中，我们评估了通过下调内源性T细胞受体（TCR）和HLA I类分子来转基因的同种异体T细胞的抗肿瘤效果以及诱导移植物抗宿主病（GVHD）的潜力，旨在开发一种"现成"的细胞制品，用于扩大基因工程T细胞的应用范围。我们使用一种新型逆转录病毒载体，通过对特异性癌胚抗原NY-ESO-1的高亲和力TCR进行siRNA靶向技术转导人淋巴细胞（siTCR载体）。这些T细胞在体外表达内源性TCR的降低，并减小对同种异体细胞的反应性。在非肥胖性糖尿病/严重综合免疫缺陷/缺陷γ链（非肥胖/SCID/γcnull）小鼠中，TCR基因转导的T细胞诱导了肿瘤的回归，而没有出现GVHD的发展。基于CRISPR/Cas9系统的慢病毒载体靶向β-2微球蛋白，在TCR基因修饰的T细胞中沉默HLA I类表达，并阻止了同种异体CD8+ T细胞的刺激而不破坏其抗肿瘤能力。本报告是首次展示了siTCR技术在预防GVHD方面的有效性。利用siTCR载体转基因的同种异体T细胞进行移植细胞治疗可能对恶性肿瘤患者的"现成"治疗有益。© 2023 The Authors. Cancer Science由日本癌症协会代表澳大利亚约翰威立出版社出版。

Adoptive immunotherapy using genetically engineered patient-derived lymphocytes to express tumor-reactive receptors is a promising treatment for malignancy. However, utilization of autologous T cells in this therapy limits the quality of gene-engineered T cells, thereby inhibiting the timely infusion of the cells into patients. In this study, we evaluated the anti-tumor efficacy and the potential to induce graft-versus-host disease (GVHD) in T cell receptor (TCR) gene-engineered allogeneic T cells that downregulate the endogenous TCR and HLA class I molecules with the aim of developing an "off-the-shelf" cell product with expanded application of genetically engineered T cells. We transduced human lymphocytes with a high-affinity TCR specific to the cancer/testis antigen NY-ESO-1 using a novel retrovirus vector with siRNAs specific to the endogenous TCR (siTCR vector). These T cells showed reduced expression of endogenous TCR and minimized reactivity to allogeneic cells in vitro. In non-obese diabetic/SCID/γcnull mice, TCR gene-transduced T cells induced tumor regression without development of GVHD. A lentivirus-based CRISPR/Cas9 system targeting β-2 microglobulin in TCR gene-modified T cells silenced the HLA class I expression and prevented allogeneic CD8+ T cell stimulation without disrupting their anti-tumor capacity. This report is the first demonstration that siTCR technology is effective in preventing GVHD. Adoptive cell therapy with allogeneic T cells engineered with siTCR vector may be useful in developing an "off-the-shelf" therapy for patients with malignancy.© 2023 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.