Full Text | PDF

Int J Biol Sci 2024; 20(5):1578-1601. doi:10.7150/ijbs.88539 This issue Cite

Research Paper

NK-92MI Cells Engineered with Anti-claudin-6 Chimeric Antigen Receptors in Immunotherapy for Ovarian Cancer

Junping Li^1,2,#, Hong Hu^1,2,#, Hui Lian^1,#, Shuo Yang^5,#, Manting Liu¹, Jinping He¹, Bihui Cao¹, Dongni Chen¹, Yuling Hu¹, Chen Zhi⁶, Yan Shen⁶, Xiaodie Ye¹, Bingjia He¹, Ming Zhao⁷, Weijun Fan⁷, Linfeng Xu⁸, Rom Leidner^9,✉, Qingde Wu^4,✉, Lili Yang^3,✉, Zhenfeng Zhang^1,✉

1. Department of Radiology; Translational Medicine Center; Guangzhou Key Laboratory for Research and Development of Nano-Biomedical Technology for Diagnosis and Therapy & Guangdong Provincial Education Department Key Laboratory of Nano-Immunoregulation Tumour Microenvironment; Central Laboratory, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, China.
2. Department of Radiology, Xiangyang No.1 People's Hospital, Hubei University of Medicine, Xiangyang 441000, China.
3. Department of Nutrition; Guangdong Provincial Key Laboratory of Food, School of Public Health, Sun Yat-sen University, Guangzhou 510080, China.
4. Department of Radiology, Shunde Chinese Medicine Hospital, the Affiliated Hospital of Traditional Chinese Medicine University of Guangzhou, Foshan 528000, China.
5. Guangzhou Municipal and Guangdong Provincial Key Laboratory of Molecular Target & Clinical Pharmacology, The NMPA and State Key Laboratory of Respiratory Disease, School of Pharmaceutical Sciences and The Fifth Affiliated Hospital, Guangzhou Medical University, Guangzhou, 510260, China.
6. Department of Pathology, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, China.
7. Minimally Invasive Interventional Division; State Key Laboratory of Oncology in South China; Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou 510060, China.
8. Department of Interventional Therapy, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510260, China.
9. Earle A. Chiles Research Institute, Providence Cancer Institute, 4805 NE Glisan St., Suite 2N35, Portland, OR 97213, USA.
^# Co-first authors.

✉ Corresponding authors: These authors jointly supervised this work: Rom Leidner, Qingde Wu, Lili Yang, Zhenfeng Zhang. E-mail: Rom.Leidnerorg (R. Leidner); wuqingde1com (Q. Wu); yangll7sysu.edu.cn (L. Yang); zhangzhfedu.cn (Z. Zhang). Lead Contact: Zhenfeng Zhang, the Second Affiliated Hospital of Guangzhou Medical University, 250 Changgangdong Road, Guangzhou 510260, China, Phone:020-34153532, E-mail address: zhangzhfedu.cn (Z. Zhang). More

Abstract

Background: The application of chimeric antigen receptor (CAR) NK cells in solid tumors is hindered by lack of tumor-specific targets and inefficient CAR-NK cell efficacy. Claudin-6 (CLDN6) has been reported to be overexpressed in ovarian cancer and may be an attractive target for CAR-NK cells immunotherapy. However, the feasibility of using anti-CLDN6 CAR-NK cells to treat ovarian cancer remains to be explored.

Methods: CLDN6 expression in primary human ovarian cancer, normal tissues and cell lines were detected by immunohistochemistry and western blot. Two types of third-generation CAR NK-92MI cells targeting CLDN6, CLDN6-CAR1 NK-92MI cells with domains containing self-activated elements (NKG2D, 2B4) and CLDN6-CAR2 NK-92MI cells with classical domains (CD28, 4-1BB) were constructed by lentivirus transfection, sorted by flow cytometry and verified by western blot and qPCR. OVCAR-3, SK-OV-3, A2780, Hey and PC-3 cells expressing the GFP and luciferase genes were transduced. Subcutaneous and intraperitoneal tumor models were established via NSG mice. The ability of CLDN6-CAR NK cells to kill CLDN6-positive ovarian cancer cells were evaluated in vitro and in vivo by live cell imaging and bioluminescence imaging.

Results: Both CLDN6-CAR1 and CLDN6-CAR2 NK-92MI cells could specifically killed CLDN6-positive ovarian cancer cells (OVCAR-3, SK-OV-3, A2780 and Hey), rather than CLDN6 negative cell (PC-3), in vitro. CLDN6-CAR1 NK-92MI cells with domains containing self-activated elements (NKG2D, 2B4) exhibited stronger cytotoxicity than CLDN6-CAR2 NK-92MI cells with classical domains (CD28, 4-1BB). Furthermore, CLDN6-CAR1 NK cells could effectively eliminate ovarian cancer cells in subcutaneous and intraperitoneal tumor models. More importantly, CAR-NK cells combined with immune checkpoint inhibitors, anti-PD-L1, could synergistically enhance the antitumor efficacy of CLDN6-targeted CAR-NK cells.

Conclusions: These results indicate that CLDN6-CAR NK cells possess strong antitumor activity and represent a promising immunotherapeutic modality for ovarian cancer.

Keywords: ovarian cancer, chimeric antigen receptor, NK cells, Claudin-6, PD-L1, PD-1

Citation styles

©2024 Ivyspring International Publisher. Terms of use