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Trained immunity-primed DLL3-targeted CAR macrophages for the eradication of small cell lung cancer
Hangjie Ying1, Yazhou Wang2, Na Li2
1Zhejiang 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.
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.
Insights
Engineered chimeric antigen receptor macrophages (CAR-Ms) targeting Delta-like ligand 3 (DLL3) show promise for treating small cell lung cancer (SCLC). Beta-glucan training further enhances CAR-M anti-tumor immunity and efficacy in preclinical models.
Area of Science:
- Immunotherapy
- Oncology
- Cellular Therapy
Background:
- Small cell lung cancer (SCLC) presents significant therapeutic challenges due to its high malignancy and limited treatment options.
- Chimeric antigen receptor macrophages (CAR-Ms) offer potential for solid tumor treatment, leveraging phagocytosis, tissue penetration, and immunomodulation.
- Delta-like ligand 3 (DLL3), a tumor-specific antigen in SCLC, is a promising target for novel therapeutic strategies.
Purpose of the Study:
- To develop and evaluate DLL3-targeted CAR-M therapy for SCLC.
- To investigate the efficacy of a β-glucan (BG)-based training protocol for enhancing CAR-M functionality.
- To assess the in vivo antitumor activity and safety of DLL3-CAR-Ms in SCLC models.
Main Methods:
- Generation of DLL3-specific CAR-Ms using lentiviral transduction of macrophages with a DLL3-targeting CAR construct.
- Establishment of a BG-based training protocol to augment CAR-M effector functions.
- Evaluation of CAR-M phagocytic and cytotoxic activities in vitro and antitumor efficacy in vivo using mouse models.
Main Results:
- DLL3-CAR-Ms demonstrated potent phagocytic and cytotoxic activity against DLL3-positive SCLC cells and effectively infiltrated tumor spheroids.
- Intravenous administration of DLL3-CAR-Ms suppressed tumor growth in both immunodeficient and immunocompetent SCLC models with no observable toxicity.
- BG training significantly enhanced CAR-M functionality, promoting sustained anti-tumor immunity, inflammatory responses, and tumor microenvironment remodeling.
Conclusions:
- DLL3-targeted CAR-Ms represent a promising therapeutic strategy for SCLC.
- BG-trained DLL3-CAR-Ms show enhanced efficacy and potential for clinical application in SCLC treatment.
- This approach may offer a viable future clinical strategy for managing solid tumors, particularly SCLC.
