Deletion of the glycan regulator SPPL3 improved allo-CAR-T fitness by reducing T cell, NK cell, and FasL-mediated killing. SPPL3 KO increased TCR/CD3 glycosylation, reduced TCR detection and antigen recognition, and diminished GvHD in humanized mice, but did not affect the CAR molecule or functionality. In 10 patients with r/r B-NHL, SPPL3KO TCRKO allo-CAR-T were safe and efficacious, without severe GvHD. However, TCR- CAR-T persistence was limited compared to TCR+ CAR-T cells. Based on this result and further preclinical studies, 3 patients received SPPL3KO TCR+ allo-CAR-T, which also showed safety and efficacy, despite TCR competence.
Contributed by Alex Najibi
ABSTRACT: Despite the success of autologous chimeric antigen receptor (CAR)-T cell therapy, achieving persistence and avoiding rejection in allogeneic settings remains challenging. We showed that signal peptide peptidase-like 3 (SPPL3) deletion enabled glycan-mediated immune evasion in primary T cells. SPPL3 deletion modified glycan profiles on T cells, restricted ligand accessibility, and reduced allogeneic immunity without compromising the functionality of anti-CD19 CAR molecules. In a phase I clinical trial, SPPL3-null, T cell receptor (TCR)-deficient anti-CD19 allogeneic CAR-T cells reached the safety primary endpoint, with grade 3 or higher cytokine release syndrome (CRS) observed in 3 out of 9 patients with relapsed/refractory B cell non-Hodgkin lymphoma (B-NHL) (ClinicalTrials.gov: NCT06014073). Reverse translational research highlighted the pivotal role of TCR in sustaining T cell persistence. We therefore evaluated the safety of SPPL3-null, TCR-sufficient CAR-T therapy on three patients with lymphoma or leukemia for compassionate care and observed no clinical signs of graft-versus-host disease. Our findings suggest glycan shielding by SPPL3 deletion is a promising direction for optimizing universal CAR-T therapies.
Author Info: (1) Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and M
Author Info: (1) Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and Modulation Research, School of Life Sciences, Peking University, Beijing 100871, China; Changping Laboratory, Beijing 102206, China. (2) School of Medicine, Nankai University, Tianjin 300071, China; Department of Bio-Therapeutic, the First Medical Center, Chinese PLA General Hospital, Beijing 100039, China. (3) Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and Modulation Research, School of Life Sciences, Peking University, Beijing 100871, China; Changping Laboratory, Beijing 102206, China. (4) Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and Modulation Research, School of Life Sciences, Peking University, Beijing 100871, China. (5) Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and Modulation Research, School of Life Sciences, Peking University, Beijing 100871, China. (6) Department of Bio-Therapeutic, the First Medical Center, Chinese PLA General Hospital, Beijing 100039, China. (7) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. (8) School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA; First School of Clinical Medicine, Peking University, Beijing 100871, China. (9) Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and Modulation Research, School of Life Sciences, Peking University, Beijing 100871, China. (10) State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China; Tianjin Institutes of Health Science, Tianjin 301600, China. (11) Department of Bio-Therapeutic, the First Medical Center, Chinese PLA General Hospital, Beijing 100039, China. (12) Department of Bio-Therapeutic, the First Medical Center, Chinese PLA General Hospital, Beijing 100039, China. (13) Department of Bio-Therapeutic, the First Medical Center, Chinese PLA General Hospital, Beijing 100039, China. (14) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. (15) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. (16) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. (17) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. (18) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. (19) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. (20) Changping Laboratory, Beijing 102206, China. (21) Changping Laboratory, Beijing 102206, China. (22) Changping Laboratory, Beijing 102206, China. (23) Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and Modulation Research, School of Life Sciences, Peking University, Beijing 100871, China. (24) Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and Modulation Research, School of Life Sciences, Peking University, Beijing 100871, China. (25) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. (26) Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and Modulation Research, School of Life Sciences, Peking University, Beijing 100871, China. (27) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. (28) Department of Bio-Therapeutic, the First Medical Center, Chinese PLA General Hospital, Beijing 100039, China. (29) Department of Bio-Therapeutic, the First Medical Center, Chinese PLA General Hospital, Beijing 100039, China. (30) First School of Clinical Medicine, Peking University, Beijing 100871, China; State Key Laboratory of Complex, Severe and Rare Diseases, Clinical Research Institute, Peking Union Medical College Hospital, Chinese Academy of Medical Science & Peking Union Medical College, Beijing 100730, China; Tsinghua-Peking University Joint Center for Life Sciences, Tsinghua University, Beijing 100084, China. (31) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. (32) EdiGene Inc., Life Science Park, Changping District, Beijing 102206, China. Electronic address: pfyuan@edigene.com. (33) Changping Laboratory, Beijing 102206, China; School of Medicine, Nankai University, Tianjin 300071, China; Department of Bio-Therapeutic, the First Medical Center, Chinese PLA General Hospital, Beijing 100039, China. Electronic address: hanwdrsw@163.com. (34) Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and Modulation Research, School of Life Sciences, Peking University, Beijing 100871, China; Changping Laboratory, Beijing 102206, China. Electronic address: wswei@pku.edu.cn.
