Multiplex Genome Editing to Generate Universal CAR T Cells Resistant to PD1 Inhibition.
Purpose:Using gene-disrupted allogeneic T cells as universal effector cells provides an alternative and potentially improves current chimeric antigen receptor (CAR) T-cell therapy against cancers and infectious diseases.Experimental Design:The CRISPR/Cas9 system has recently emerged as a simple and efficient way for multiplex genome engineering. By combining lentiviral delivery of CAR and electro-transfer of Cas9 mRNA and gRNAs targeting endogenous TCR, β-2 microglobulin (B2M) and PD1 simultaneously, to generate gene-disrupted allogeneic CAR T cells deficient of TCR, HLA class I molecule and PD1.Results:The CRISPR gene-edited CAR T cells showed potent antitumor activities, bothin vitroand in animal models and were as potent as non-gene-edited CAR T cells. In addition, the TCR and HLA class I double deficient T cells had reduced alloreactivity and did not cause graft-versus-host disease. Finally, simultaneous triple genome editing by adding the disruption of PD1 led to enhancedin vivoantitumor activity of the gene-disrupted CAR T cells.Conclusions:Gene-disrupted allogeneic CAR and TCR T cells could provide an alternative as a universal donor to autologous T cells, which carry difficulties and high production costs. Gene-disrupted CAR and TCR T cells with disabled checkpoint molecules may be potent effector cells against cancers and infectious diseases.Clin Cancer Res; 23(9); 2255-66. ©2016 AACR.
Abramson Cancer Center, University of Pennsylvania, Philadelphia, Pennsylvania.
Jiangtao
J
Ren
Abramson Cancer Center, University of Pennsylvania, Philadelphia, Pennsylvania.
Xiaojun
X
Liu
Abramson Cancer Center, University of Pennsylvania, Philadelphia, Pennsylvania.
Chongyun
C
Fang
Abramson Cancer Center, University of Pennsylvania, Philadelphia, Pennsylvania.
Shuguang
S
Jiang
Abramson Cancer Center, University of Pennsylvania, Philadelphia, Pennsylvania. Yangbing@exchange.upenn.edu cjune@exchange.upenn.edu.|Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.|Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.
Carl H
CH
June
Abramson Cancer Center, University of Pennsylvania, Philadelphia, Pennsylvania. Yangbing@exchange.upenn.edu cjune@exchange.upenn.edu.|Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.|Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.
Yangbing
Y
Zhao
D064113:CRISPR-Cas Systems; D000072669:Gene Editing; D006086:Graft vs Host Disease; D006801:Humans; D016219:Immunotherapy, Adoptive; D008213:Lymphocyte Activation / Q000276:immunology; D009369:Neoplasms / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D061026:Programmed Cell Death 1 Receptor / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics*; D011948:Receptors, Antigen, T-Cell / Q000172:deficiency / Q000235:genetics / Q000276:immunology; D016511:Severe Combined Immunodeficiency / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
C105992:PDCD1 protein, human; D061026:Programmed Cell Death 1 Receptor; D011948:Receptors, Antigen, T-Cell