Trained immunity-primed DLL3-targeted CAR macrophages for the eradication of small cell lung cancer

Hangjie Ying; Yazhou Wang; Na Li; Wenshuo Ma; Xin Shou; Zhiyun Liu; Changjiang Chen; Zhenzhen Dong; Hung-Wing Li; Junfeng Zhang; Ying Jin; Sergei Krasny; Liyun Shi

doi:10.1016/j.jare.2026.05.047

Trained immunity-primed DLL3-targeted CAR macrophages for the eradication of small cell lung cancer

J Adv Res. 2026 Jun 1:S2090-1232(26)00443-1. doi: 10.1016/j.jare.2026.05.047. Online ahead of print.

Authors

Hangjie Ying¹, Yazhou Wang², Na Li², Wenshuo Ma², Xin Shou³, Zhiyun Liu³, Changjiang Chen⁴, Zhenzhen Dong³, Hung-Wing Li⁵, Junfeng Zhang², Ying Jin⁶, Sergei Krasny⁷, Liyun Shi⁸

Affiliations

¹ Zhejiang Cancer Hospital, Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences, Hangzhou, Zhejiang 310022, China; Key Laboratory of Artificial Organs and Computational Medicine in Zhejiang Province, Institute of Translational Medicine, Zhejiang Shuren University, Hangzhou, Zhejiang 310015, China; Department of Immunology, School of Medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, China.
² Department of Immunology, School of Medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, China.
³ Key Laboratory of Artificial Organs and Computational Medicine in Zhejiang Province, Institute of Translational Medicine, Zhejiang Shuren University, Hangzhou, Zhejiang 310015, China.
⁴ Key Laboratory of Artificial Organs and Computational Medicine in Zhejiang Province, Institute of Translational Medicine, Zhejiang Shuren University, Hangzhou, Zhejiang 310015, China; Department of Immunology, School of Medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, China.
⁵ Department of Chemistry, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
⁶ Zhejiang Cancer Hospital, Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences, Hangzhou, Zhejiang 310022, China; Wenzhou Medical University, Wenzhou, Zhejiang 325035, China. Electronic address: jinying@zjcc.org.cn.
⁷ N. N. Alexandrov National Cancer Centre of Belarus, Minsk, Belarus.
⁸ Key Laboratory of Artificial Organs and Computational Medicine in Zhejiang Province, Institute of Translational Medicine, Zhejiang Shuren University, Hangzhou, Zhejiang 310015, China; Yuhang Institute of Medical Science Innovation and Transformation, Hangzhou 311112, China. Electronic address: sly0202@zjsru.edu.cn.

PMID: 42229717
DOI: 10.1016/j.jare.2026.05.047

Abstract

Background: Small cell lung cancer (SCLC) is highly malignant with limited treatment options. Chimeric antigen receptor macrophages (CAR-Ms) show potential for solid tumor therapy due to their phagocytic activity, tissue penetration, and immunomodulatory functions, but their application in SCLC remains unexplored. Delta-like ligand 3 (DLL3), a SCLC-specific membrane antigen, represents a promising therapeutic target. Here, we developed a DLL3-targeted CAR-M therapy and an enhanced strategy for SCLC immunotherapy.

Methods: DLL3-specific CAR-Ms were generated by introducing a CAR construct (DLL3-ScFv-CD8-CD3ζ) into murine and human macrophages via lentiviral transduction. A β-glucan (BG)-based training protocol was established to enhance CAR-M functionality. Phagocytic and cytotoxic activities were evaluated by flow cytometry and bioluminescence assays, and in vivo antitumor efficacy was assessed in immunodeficient and immunocompetent mouse models.

Results: Engineered CAR-Ms exhibited potent phagocytic and cytotoxic activity against DLL3-positive cells and effectively infiltrated and eliminated tumor spheroids in 3D culture systems. Intravenously administered CAR-Ms suppressed DLL3-positive lung cancer growth in both immunodeficient and immunocompetent models without discernible toxicity. Importantly, BG training enhanced CAR-M functionality by conferring sustained anti-tumor immunity, amplifying inflammatory and interferon pathway activation, and remodeling the tumor microenvironment through epigenetic and metabolic reprogramming. These findings establish BG-trained, DLL3-targeting CAR-Ms as a promising therapeutic approach for SCLC.

Conclusion: Anti-DLL3 CAR-Ms demonstrate significant potential for solid tumor treatment and may offer a viable clinical strategy for SCLC in the future.

Keywords: Chimeric antigen receptor; Delta-like protein 3; Macrophage; Small cell lung cancer; Trained immunity.