Experimental Hematology Unit, IRCCS San Raffaele Scientific Institute, Milan, Italy.
Center for Omics Sciences, IRCCS San Raffaele Scientific Institute, Milan, Italy.
Gut. 2023 Oct;72(10):1887-1903. doi: 10.1136/gutjnl-2022-328042. Epub 2023 Jun 30.
Colorectal tumours are often densely infiltrated by immune cells that have a role in surveillance and modulation of tumour progression but are burdened by immunosuppressive signals, which might vary from primary to metastatic stages. Here, we deployed a multidimensional approach to unravel the T-cell functional landscape in primary colorectal cancers (CRC) and liver metastases, and genome editing tools to develop CRC-specific engineered T cells.
We paired high-dimensional flow cytometry, RNA sequencing and immunohistochemistry to describe the functional phenotype of T cells from healthy and neoplastic tissue of patients with primary and metastatic CRC and we applied lentiviral vectors (LV) and CRISPR/Cas9 genome editing technologies to develop CRC-specific cellular products.
We found that T cells are mainly localised at the front edge and that tumor-infiltrating T cells co-express multiple inhibitory receptors, which largely differ from primary to metastatic sites. Our data highlighted CD39 as the major driver of exhaustion in both primary and metastatic colorectal tumours. We thus simultaneously redirected T-cell specificity employing a novel T-cell receptor targeting HER-2 and disrupted the endogenous TCR genes (TCR editing (TCR)) and the CD39 encoding gene (), thus generating TCRENTPD1HER-2-redirected lymphocytes. We showed that the absence of CD39 confers to HER-2-specific T cells a functional advantage in eliminating HER-2 patient-derived organoids and .
HER-2-specific CD39 disrupted engineered T cells are promising advanced medicinal products for primary and metastatic CRC.
结直肠肿瘤常被浸润大量免疫细胞，这些细胞在肿瘤进展的监测和调节中发挥作用，但也受到免疫抑制信号的影响，这些信号可能因原发灶和转移灶而有所不同。在这里，我们采用多维方法来揭示原发性结直肠癌（CRC）和肝转移灶中 T 细胞的功能图谱，并利用基因组编辑工具开发 CRC 特异性工程 T 细胞。
我们将高维流式细胞术、RNA 测序和免疫组织化学相结合，描述了来自原发性和转移性 CRC 患者健康和肿瘤组织中 T 细胞的功能表型，并应用慢病毒载体（LV）和 CRISPR/Cas9 基因组编辑技术来开发 CRC 特异性细胞产品。
我们发现 T 细胞主要定位于前缘，肿瘤浸润 T 细胞共表达多种抑制性受体，这些受体在原发灶和转移灶之间存在很大差异。我们的数据突出了 CD39 作为原发性和转移性结直肠肿瘤中衰竭的主要驱动因素。因此，我们同时采用了一种新型靶向 HER-2 的 T 细胞受体，并破坏内源性 TCR 基因（TCR 编辑（TCR））和 CD39 编码基因（），从而生成 TCRENTPD1HER-2 重定向的淋巴细胞，同时重新定向 T 细胞的特异性。我们表明，CD39 的缺失赋予了 HER-2 特异性 T 细胞在消除 HER-2 患者衍生类器官和异种移植瘤中的功能优势。
HER-2 特异性 CD39 破坏的工程 T 细胞是治疗原发性和转移性 CRC 的有前途的先进药物。
