嵌合抗原受体（CAR）-NK92细胞在体外以及卵巢癌小鼠异种移植模型中对胶质母细胞瘤、乳腺癌和胰腺癌有效。

Chimeric Antigen Receptor (CAR)-NK92 cells effective against glioblastoma, breast- and pancreatic cancer in vitro and in a murine xenograft model of ovarian cancer.

作者信息

Boulifa Abdelhadi, Franzén Alexander Sebastian, Raftery Martin J, Radecke Clarissa, Pecher Gabriele

机构信息

Berlin Institute of Health at Charité- Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany.

Competence Center of Immuno-Oncology and Translational Cell Therapy (KITZ), Department of Hematology, Oncology and Tumor Immunology, CCM, Charité- Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Charitéplatz 1, 10117, Berlin, Germany.

出版信息

Cancer Cell Int. 2025 Jul 11;25(1):260. doi: 10.1186/s12935-025-03865-0.

DOI:10.1186/s12935-025-03865-0

PMID:40646591

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12255008/

Abstract

BACKGROUND

Aggressive tumors such as glioblastoma, breast, ovarian and pancreatic cancer have low survival rates and new therapies are urgently needed. One potential target is CD44v6, a splice variant of CD44 that is associated with poor prognosis. Recently, NK cells expressing CAR molecules have shown promise in combining specific targeting of solid tumors with a low risk of side effects. The aim of the current study is to explore the efficacy of the CD44v6-CAR construct expressed in the NK cell line NK92 against solid tumors both in vitro and in vivo.

METHODS

Flow cytometry was used to evaluate the expression of CD44v6 on glioblastoma, breast, ovarian and pancreatic cancer cell lines. In order to investigate the efficacy of CD44v6-CAR-NK92 against these solid tumors in 2D and 3D models, cytotoxicity was measured using a luminescent cell viability assay. Additionally, we assessed the levels of IFN-γ in cell culture supernatants using an ELISA method. Finally, we evaluated our therapeutic in vivo using a xenografted murine model of ovarian cancer through bioluminescent imaging.

RESULTS

CD44v6-CAR-NK92 cells exhibit specific cytotoxicity against glioblastoma, breast, ovarian and pancreatic cancer after 24 h compared to the control, both in 2D and 3D models. Furthermore, the activity of CD44v6-CAR-NK92 was validated by quantifying specific cytokine release in response to target cells. Finally, we could show that CD44v6-CAR-NK92 was effective in reducing tumor burden in a xenografted murine model of ovarian cancer.

CONCLUSION

Our results demonstrate that CD44v6-CAR-NK92 cells could be an attractive therapeutic agent for the treatment of solid tumors.

摘要

背景

胶质母细胞瘤、乳腺癌、卵巢癌和胰腺癌等侵袭性肿瘤生存率低，迫切需要新的治疗方法。一个潜在靶点是CD44v6，它是CD44的一种剪接变体，与预后不良相关。最近，表达嵌合抗原受体（CAR）分子的自然杀伤（NK）细胞在将实体瘤的特异性靶向与低副作用风险相结合方面显示出前景。本研究的目的是探讨在NK细胞系NK92中表达的CD44v6-CAR构建体在体外和体内对实体瘤的疗效。

方法

采用流式细胞术评估胶质母细胞瘤、乳腺癌、卵巢癌和胰腺癌细胞系上CD44v6的表达。为了研究CD44v6-CAR-NK92在二维和三维模型中对这些实体瘤的疗效，使用发光细胞活力测定法测量细胞毒性。此外，我们采用酶联免疫吸附测定法评估细胞培养上清液中γ干扰素（IFN-γ）的水平。最后，我们通过生物发光成像，利用卵巢癌异种移植小鼠模型评估我们的体内治疗效果。

结果

与对照组相比，在二维和三维模型中，CD44v6-CAR-NK92细胞在24小时后对胶质母细胞瘤、乳腺癌、卵巢癌和胰腺癌表现出特异性细胞毒性。此外，通过量化对靶细胞反应时特定细胞因子的释放，验证了CD44v6-CAR-NK92的活性。最后，我们可以证明CD44v6-CAR-NK92在卵巢癌异种移植小鼠模型中有效减轻肿瘤负担。