Lei and Yu et al. engineered iPSC-derived macrophages (iMACs) with a CAR containing a toll-like receptor 4 intracellular toll/IL-1R (TIR) domain to provide an orthogonal signal for macrophage polarization. Co-integration of TIR and CD3ζ into CAR-iMACs enhanced antigen-dependent antitumor efficacy, both in vitro and in vivo. scRNAseq analysis showed that TIR promoted the M1-like pro-inflammatory state and suppressed the M2-like state in an NF-κB pathway-dependent manner. CAR engagement with tumor antigens activated CAR-iMACs, leading to TNF secretion that mediated tumor cell apoptosis, followed by clearance of apoptotic bodies through efferocytosis.
Contributed by Shishir Pant
ABSTRACT: Chimeric antigen receptor (CAR) T cell therapies have successfully treated hematological malignancies. Macrophages have also gained attention as an immunotherapy owing to their immunomodulatory capacity and ability to infiltrate solid tumors and phagocytize tumor cells. The first-generation CD3ζ-based CAR-macrophages could phagocytose tumor cells in an antigen-dependent manner. Here we engineered induced pluripotent stem cell-derived macrophages (iMACs) with toll-like receptor 4 intracellular toll/IL-1R (TIR) domain-containing CARs resulting in a markedly enhanced antitumor effect over first-generation CAR-macrophages. Moreover, the design of a tandem CD3ζ-TIR dual signaling CAR endows iMACs with both target engulfment capacity and antigen-dependent M1 polarization and M2 resistance in a nuclear factor kappa B (NF-κB)-dependent manner, as well as the capacity to modulate the tumor microenvironment. We also outline a mechanism of tumor cell elimination by CAR-induced efferocytosis against tumor cell apoptotic bodies. Taken together, we provide a second-generation CAR-iMAC with an ability for orthogonal phagocytosis and polarization and superior antitumor functions in treating solid tumors relative to first-generation CAR-macrophages.
Author Info: (1) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang Universi
Author Info: (1) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. Liangzhu Laboratory, Zhejiang University, Hangzhou, China. Institute of Hematology, Zhejiang University, Hangzhou, China. CellOrigin Inc, Hangzhou, China. (2) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. School of Basic Medical Sciences, Nanchang University, Nanchang, China. (3) Department of Surgical Oncology, Second Affiliated Hospital, Zhejiang University School of Medicine, Zhejiang University, Hangzhou, China. (4) Liangzhu Laboratory, Zhejiang University, Hangzhou, China. (5) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. Liangzhu Laboratory, Zhejiang University, Hangzhou, China. (6) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. Liangzhu Laboratory, Zhejiang University, Hangzhou, China. (7) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. Liangzhu Laboratory, Zhejiang University, Hangzhou, China. (8) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. (9) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. Institute of Hematology, Zhejiang University, Hangzhou, China. (10) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. Liangzhu Laboratory, Zhejiang University, Hangzhou, China. (11) Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, China. (12) Liangzhu Laboratory, Zhejiang University, Hangzhou, China. (13) Liangzhu Laboratory, Zhejiang University, Hangzhou, China. (14) Eye Center of the Second Affiliated Hospital, Zhejiang University School of Medicine, Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, China. (15) MOE Frontier Science Center for Brain Science and Brain-machine Integration, State Key Laboratory of Brain-machine Intelligence, Zhejiang University, Hangzhou, China. NHC and CAMS Key Laboratory of Medical Neurobiology, Zhejiang University, Hangzhou, China. (16) Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, China. (17) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. Liangzhu Laboratory, Zhejiang University, Hangzhou, China. Institute of Hematology, Zhejiang University, Hangzhou, China. (18) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. (19) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. Liangzhu Laboratory, Zhejiang University, Hangzhou, China. (20) Liangzhu Laboratory, Zhejiang University, Hangzhou, China. (21) Department of Genetics and Wyss Institute for Biologically Inspired Engineering, Harvard Medical School, Boston, MA, USA. (22) Liangzhu Laboratory, Zhejiang University, Hangzhou, China. (23) MOE Frontier Science Center for Brain Science and Brain-machine Integration, State Key Laboratory of Brain-machine Intelligence, Zhejiang University, Hangzhou, China. NHC and CAMS Key Laboratory of Medical Neurobiology, Zhejiang University, Hangzhou, China. (24) Center for Stem Cell and Regenerative Medicine, Department of Basic Medical Sciences, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China. zhgene@zju.edu.cn. Liangzhu Laboratory, Zhejiang University, Hangzhou, China. zhgene@zju.edu.cn. Institute of Hematology, Zhejiang University, Hangzhou, China. zhgene@zju.edu.cn. Center of Gene and Cell Therapy and Genome Medicine of Zhejiang Province, Hangzhou, China. zhgene@zju.edu.cn.
