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CRISPR/Cas9-mediated PD-1 Disruption Enhances Anti-tumor Efficacy Of Human Chimeric Antigen Receptor T Cells

Levi J. Rupp, Kathrin Schumann, Kole T. Roybal, Rachel E. Gate, Chun Jimmie Ye, Wendell A Lim, Alexander Marson
Published 2017 · Biology, Medicine
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Immunotherapies with chimeric antigen receptor (CAR) T cells and checkpoint inhibitors (including antibodies that antagonize programmed cell death protein 1 [PD-1]) have both opened new avenues for cancer treatment, but the clinical potential of combined disruption of inhibitory checkpoints and CAR T cell therapy remains incompletely explored. Here we show that programmed death ligand 1 (PD-L1) expression on tumor cells can render human CAR T cells (anti-CD19 4-1BBζ) hypo-functional, resulting in impaired tumor clearance in a sub-cutaneous xenograft model. To overcome this suppressed anti-tumor response, we developed a protocol for combined Cas9 ribonucleoprotein (Cas9 RNP)-mediated gene editing and lentiviral transduction to generate PD-1 deficient anti-CD19 CAR T cells. Pdcd1 (PD-1) disruption augmented CAR T cell mediated killing of tumor cells in vitro and enhanced clearance of PD-L1+ tumor xenografts in vivo. This study demonstrates improved therapeutic efficacy of Cas9-edited CAR T cells and highlights the potential of precision genome engineering to enhance next-generation cell therapies.
This paper references
10.1056/NEJMoa1200690
Safety, activity, and immune correlates of anti-PD-1 antibody in cancer.
S. Topalian (2012)
10.1073/pnas.0813101106
Control of large, established tumor xenografts with genetically retargeted human T cells containing CD28 and CD137 domains
C. Carpenito (2009)
10.1158/1535-7163.MCT-14-0983
PD-L1 Expression as a Predictive Biomarker in Cancer Immunotherapy
S. Patel (2015)
10.1126/science.aaa6204
T cell exclusion, immune privilege, and the tumor microenvironment
Johanna A. Joyce (2015)
10.1016/j.ccell.2015.09.004
Structural Design of Engineered Costimulation Determines Tumor Rejection Kinetics and Persistence of CAR T Cells.
Zeguo Zhao (2015)
A Genome - wide CRISPR Screen in Primary Immune
O. Parnas
10.1158/0008-5472.CAN-14-3321
Multiplex Genome-Edited T-cell Manufacturing Platform for "Off-the-Shelf" Adoptive T-cell Immunotherapies.
Laurent Poirot (2015)
10.1126/scitranslmed.aad5222
Early memory phenotypes drive T cell proliferation in patients with pediatric malignancies
Nathan Singh (2016)
10.1146/annurev-immunol-032713-120136
Adoptive immunotherapy for cancer or viruses.
M. Maus (2014)
10.1128/MCB.25.21.9543-9553.2005
CTLA-4 and PD-1 Receptors Inhibit T-Cell Activation by Distinct Mechanisms
R. Parry (2005)
T cells engineered to express a CD19-specific chimericantigen-receptor and eliminate expression of endogenous TCR
Torikai (2012)
10.1126/science.aaa4967
Adoptive cell transfer as personalized immunotherapy for human cancer
S. A. Rosenberg (2015)
10.1016/j.cell.2013.04.025
One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering
H. Wang (2013)
10.1073/pnas.1512503112
Generation of knock-in primary human T cells using Cas9 ribonucleoproteins
K. Schumann (2015)
10.1016/j.cell.2015.06.059
A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks
Oren Parnas (2015)
10.1126/science.aab4077
Remote control of therapeutic T cells through a small molecule–gated chimeric receptor
Chia-Yung Wu (2015)
10.1126/scitranslmed.3006597
PD-1– and CTLA-4–Based Inhibitory Chimeric Antigen Receptors (iCARs) Divert Off-Target Immunotherapy Responses
Victor Dmitri Fedorov (2013)
10.1016/j.coi.2015.06.015
Synthetic biology approaches to engineer T cells.
Chia-Yung Wu (2015)
10.1038/nature04444
Restoring function in exhausted CD8 T cells during chronic viral infection
D. Barber (2006)
10.1056/NEJMoa1200694
Safety and activity of anti-PD-L1 antibody in patients with advanced cancer.
J. Brahmer (2012)
10.1182/blood-2012-01-405365
A foundation for universal T-cell based immunotherapy: T cells engineered to express a CD19-specific chimeric-antigen-receptor and eliminate expression of endogenous TCR.
Hiroki Torikai (2012)
10.1182/blood-2013-10-535245
ICOS-based chimeric antigen receptors program bipolar TH17/TH1 cells.
Sonia Guedan (2014)
10.1016/j.cell.2015.08.052
Melanoma Cell-Intrinsic PD-1 Receptor Functions Promote Tumor Growth
Sonja Kleffel (2015)
10.1016/B978-0-12-801185-0.00001-5
In vitro enzymology of Cas9.
Carolin Anders (2014)
10.1016/S0140-6736(18)31559-9
Melanoma
D. Schadendorf (2018)
10.1038/nbt.3290
Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells
Ayal Hendel (2015)
10.1038/nbt.3117
GUIDE-Seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases
S. Tsai (2015)
10.1016/S1525-0016(16)33539-0
731. High-Fidelity CRISPR-Cas9 Nucleases with No Detectable Genome-Wide Off-Target Effects
Benjamin P. Kleinstiver (2016)
10.1158/1078-0432.CCR-13-0458
Anti-PD-1 Antibody Therapy Potently Enhances the Eradication of Established Tumors By Gene-Modified T Cells
L. B. John (2013)
10.1056/NEJMoa1407222
Chimeric antigen receptor T cells for sustained remissions in leukemia.
Shannon L Maude (2014)
10.1084/jem.20142237
Genetic absence of PD-1 promotes accumulation of terminally differentiated exhausted CD8+ T cells
Pamela M. Odorizzi (2015)
10.1038/gt.2014.26
TALEN-mediated editing of endogenous T-cell receptors facilitates efficient reprogramming of T lymphocytes by lentiviral gene transfer
Belinda Berdien (2014)
10.1126/scitranslmed.3003130
The PDL1-PD1 Axis Converts Human TH1 Cells into Regulatory T Cells
S. Amarnath (2011)
10.1016/j.it.2015.02.008
Overcoming T cell exhaustion in infection and cancer.
Kristen E. Pauken (2015)
Releasing the Brake on the Immune System: The PD-1 Strategy for Hematologic Malignancies.
L. Bryan (2015)
10.1016/j.cell.2015.08.016
Metabolic Competition in the Tumor Microenvironment Is a Driver of Cancer Progression
Chih-Hao Chang (2015)
10.1146/annurev.immunol.26.021607.090331
PD-1 and its ligands in tolerance and immunity.
M. Keir (2008)
10.1056/NEJMoa1411087
PD-1 blockade with nivolumab in relapsed or refractory Hodgkin's lymphoma.
S. Ansell (2015)
to Dissect Regulatory Networks
C. Anders (2015)
10.1126/science.aad5227
Rationally engineered Cas9 nucleases with improved specificity
I. Slaymaker (2016)
10.1016/j.stem.2014.10.004
Efficient ablation of genes in human hematopoietic stem and effector cells using CRISPR/Cas9.
P. Mandal (2014)



This paper is referenced by
10.1016/j.intimp.2019.01.010
Chimeric antigen receptor T cell therapy and other therapeutics for malignancies: Combination and opportunity.
Luyao Wang (2019)
10.1007/s40259-019-00368-z
Chimeric Antigen Receptor-T Cells for Targeting Solid Tumors: Current Challenges and Existing Strategies
Lorraine Springuel (2019)
10.1007/978-1-0716-0290-4_23
Engineering T Cells Using CRISPR/Cas9 for Cancer Therapy.
Xingying Zhang (2020)
10.1111/imm.13094
Genetic Reprogramming For NK cell Cancer Immunotherapy with CRISPR/Cas9.
Lukman O. Afolabi (2019)
10.1126/scitranslmed.aaw2293
Supercharging adoptive T cell therapy to overcome solid tumor–induced immunosuppression
Sherly Mardiana (2019)
10.2991/CHI.D.190219.001
Relapse and Resistance to CAR-T Cells and Blinatumomab in Hematologic Malignancies
Elad Jacoby (2019)
10.1002/cyto.b.21880
New directions in chimeric antigen receptor T cell [CAR-T] therapy and related flow cytometry.
Elena Maryamchik (2020)
10.1080/14712598.2017.1339687
Practical considerations for chimeric antigen receptor design and delivery
Robyn A. A. Oldham (2017)
10.1155/2018/3797214
CRISPR/Cas9 System: A Bacterial Tailor for Genomic Engineering
Bilal Ahmad Lone (2018)
10.12659/MSM.910903
Programmed Death Receptor 1 (PD1) Knockout and Human Telomerase Reverse Transcriptase (hTERT) Transduction Can Enhance Persistence and Antitumor Efficacy of Cytokine-Induced Killer Cells Against Hepatocellular Carcinoma
Kanghua Huang (2018)
10.1155/2017/8960236
The Impact of CRISPR/Cas9 Technology on Cardiac Research: From Disease Modelling to Therapeutic Approaches
Benedetta M Motta (2017)
Delineating the impact of binding-domain affinity and kinetic properties on Chimeric Antigen Receptor T-cell function
Anne Marijn Kramer (2017)
10.1038/s41587-019-0137-8
Gene editing for immune cell therapies
Stefanie R Bailey (2019)
10.15825/1995-1191-2018-3-95-104
Аутотрансплантация Т-лимфоцитов как инструмент для антиген-специфической иммунотерапии онкологических заболеваний
А. Ю. Лупатов (2018)
10.1016/j.yapd.2019.04.001
Gene Therapy: Current Applications and Future Possibilities.
David H.M. Steffin (2019)
10.1021/acschembio.7b00680
The Promise and Challenge of In Vivo Delivery for Genome Therapeutics.
Ross C Wilson (2018)
Gene editing in T-cells and T-cell targets
Angharad Lloyd (2016)
10.1080/2162402X.2016.1273302
CAR T-cell intrinsic PD-1 checkpoint blockade: A two-in-one approach for solid tumor immunotherapy
Nan Chen (2017)
10.1007/978-3-319-62941-4_1
Precision Medicine in Lung Cancer
Keith M. Kerr (2018)
10.1038/s41416-018-0325-1
Teaching an old dog new tricks: next-generation CAR T cells
Nicholas Tokarew (2018)
10.1007/s00262-018-2281-2
CRISPR/Cas9-mediated PD-1 disruption enhances human mesothelin-targeted CAR T cell effector functions
Wanghong Hu (2018)
10.3390/ijms19092721
In Vivo Genome Editing as a Therapeutic Approach
Beatrice Xuan Ho (2018)
10.24294/ti.v2.i3.1064
CAR-T Therapy for Solid Tumors: Development of New Strategies
Samuel D. Bernal (2019)
10.1002/cam4.2430
Research progress and design optimization of CAR‐T therapy for pancreatic ductal adenocarcinoma
Tianjiao Li (2019)
10.1126/science.aat5011
CRISPR-Cas guides the future of genetic engineering
G. Knott (2018)
10.1038/NATREVMATS.2017.56
Tools for translation: non-viral materials for therapeutic mRNA delivery
Khalid A Hajj (2017)
10.1101/384776
Genome-wide CRISPR Screens in Primary Human T Cells Reveal Key Regulators of Immune Function
Eric Shifrut (2018)
10.1007/s12307-018-0208-2
Matrix Metalloproteinase 8: Could it Benefit the CAR-T Cell Therapy of Solid Tumors?- a- Commentary on Therapeutic Potential
Alireza Mardomi (2018)
10.1007/978-3-030-38862-1
Tumor Microenvironment
Steven T Rosen (2020)
10.1126/science.aba9844
Knocking out barriers to engineered cell activity
Jennifer R. Hamilton (2020)
10.1038/s41467-020-17011-z
Oncolytic virus-derived type I interferon restricts CAR T cell therapy
Laura Evgin (2020)
10.3389/fimmu.2019.00128
CAR T Cells for Solid Tumors: New Strategies for Finding, Infiltrating, and Surviving in the Tumor Microenvironment
Marina Martinez (2019)
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