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Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection
Open accessAbstract
Chimeric antigen receptors (CARs) are synthetic receptors that redirect and reprogram T cells to mediate tumour rejection. The most successful CARs used to date are those targeting CD19 (ref. 2), which offer the prospect of complete remission in patients with chemorefractory or relapsed B-cell malignancies. CARs are typically transduced into the T cells of a patient using γ-retroviral vectors or other randomly integrating vectors, which may result in clonal expansion, oncogenic transformation, variegated transgene expression and transcriptional silencing. Recent advances in genome editing enable efficient sequence-specific interventions in human cells, including targeted gene delivery to the CCR5 and AAVS1 loci. Here we show that directing a CD19-specific CAR to the T-cell receptor α constant (TRAC) locus not only results in uniform CAR expression in human peripheral blood T cells, but also enhances T-cell potency, with edited cells vastly outperforming conventionally generated CAR T cells in a mouse model of acute lymphoblastic leukaemia. We further demonstrate that targeting the CAR to the TRAC locus averts tonic CAR signalling and establishes effective internalization and re-expression of the CAR following single or repeated exposure to antigen, delaying effector T-cell differentiation and exhaustion. These findings uncover facets of CAR immunobiology and underscore the potential of CRISPR/Cas9 genome editing to advance immunotherapies.
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“AAV6 homologous donor DNA templates are an efficient way to deliver donor templates for sensitive primary immune cells, including HSPCs and T-Cells15,35–40.”
From A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells (2018) · cites this paper as [39]
Experimental — accuracy not yet measuredIn the citing paper
“More recent work has revealed potential advantages of targeted integrations of antigen receptor genes into defined genomic loci (Eyquem et al., 2017), which can be achieved with programmable nucleases such as CRISPR-Cas9 along with viral or non-viral knock-in templates (Gwiazda et al., 2016; Eyquem et al., 2017; Roth et al., 2018).”
From Pooled Knockin Targeting for Genome Engineering of Cellular Immunotherapies (2020) · cites this paper as Eyquem et al., 2017
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Citations
Cites
Molecular and cellular insights into T cell exhaustion
Wherry, E. J., & Kurachi, M. (2015) · Nature Reviews Immunology · doi:10.1038/nri3862 · Cited by 4,652
Brentjens, R. J., Davila, M. L., Riviere, I., Park, J., Wang, X., Cowell, L. G., Bartido, S., Stefanski, J., Taylor, C., Olszewska, M., et al. (2013) · Science Translational Medicine · doi:10.1126/scitranslmed.3005930 · Cited by 2,063
Self-citationBlackburn, S. D., Shin, H., Haining, W. N., Zou, T., Workman, C. J., Polley, A., Betts, M. R., Freeman, G. J., Vignali, D. A. A., & Wherry, E. J. (2009) · Nature Immunology · doi:10.1038/ni.1679 · Cited by 2,062
A human memory T cell subset with stem cell–like properties
Gattinoni, L., Lugli, E., Ji, Y., Pos, Z., Paulos, C. M., Quigley, M. F., Almeida, J. R., Gostick, E., Yu, Z., Carpenito, C., et al. (2011) · Nature Medicine · doi:10.1038/nm.2446 · Cited by 1,974
Long, A. H., Haso, W. M., Shern, J. F., Wanhainen, K. M., Murgai, M., Ingaramo, M., Smith, J. P., Walker, A. J., Kohler, M. E., Venkateshwara, V. R., et al. (2015) · Nature Medicine · doi:10.1038/nm.3838 · Cited by 1,815
Human T-lymphocyte cytotoxicity and proliferation directed by a single chimeric TCRζ /CD28 receptor
Maher, J. F., Brentjens, R. J., Gunset, G., Rivière, I., & Sadelain, M. (2002) · Nature Biotechnology · doi:10.1038/nbt0102-70 · Cited by 1,031
Self-citationBiology and Applications of CRISPR Systems: Harnessing Nature’s Toolbox for Genome Engineering
Wright, A. V., Nuñez, J. K., & Doudna, J. A. (2016) · Cell · doi:10.1016/j.cell.2015.12.035 · Cited by 989
Sommermeyer, D., Hudecek, M., Kosasih, P. L., Gogishvili, T., Maloney, D. G., Turtle, C. J., & Riddell, S. R. (2015) · Leukemia · doi:10.1038/leu.2015.247 · Cited by 890
Zhao, Z., Condomines, M., van der Stegen, S. J. C., Perna, F., Kloss, C. C., Gunset, G., Plotkin, J., & Sadelain, M. (2015) · Cancer Cell · doi:10.1016/j.ccell.2015.09.004 · Cited by 843
Self-citationTolerance and exhaustion: defining mechanisms of T cell dysfunction
Schietinger, A., & Greenberg, P. D. (2013) · Trends in Immunology · doi:10.1016/j.it.2013.10.001 · Cited by 704
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A guide to cancer immunotherapy: from T cell basic science to clinical practice
Waldman, A. D., Fritz, J. M., & Lenardo, M. J. (2020) · Nature Reviews Immunology · doi:10.1038/s41577-020-0306-5 · Cited by 4,154
Approaches to treat immune hot, altered and cold tumours with combination immunotherapies
Galon, J., & Bruni, D. (2019) · Nature Reviews Drug Discovery · doi:10.1038/s41573-018-0007-y · Cited by 3,489
Car t cell immunotherapy for human cancer
June, C. H., O’Connor, R. S., Kawalekar, O. U., Ghassemi, S., & Milone, M. C. (2018) · Science · doi:10.1126/science.aar6711 · Cited by 3,207
Chimeric Antigen Receptor Therapy
June, C. H., & Sadelain, M. (2018) · New England Journal of Medicine · doi:10.1056/nejmra1706169 · Cited by 2,411
Self-citationEngineering strategies to overcome the current roadblocks in CAR T cell therapy
Rafiq, S., Hackett, C. S., & Brentjens, R. J. (2019) · Nature Reviews Clinical Oncology · doi:10.1038/s41571-019-0297-y · Cited by 1,511
“…gene construct into the TRAC locus has also been achieved using CRISPR– Cas9 in preclinical models, thereby knocking out the TCR and inserting the CAR in one step 238 ; this feat has also been achieved using an engineered I-CreI homing endonuclease and an adeno-associated virus donor template 239 .”
MethodThe next generation of CRISPR–Cas technologies and applications
Pickar-Oliver, A., & Gersbach, C. A. (2019) · Nature Reviews Molecular Cell Biology · doi:10.1038/s41580-019-0131-5 · Cited by 1,459
CRISPR-Cas guides the future of genetic engineering
Knott, G. J., & Doudna, J. A. (2018) · Science · doi:10.1126/science.aat5011 · Cited by 1,410
Dunbar, C. E., High, K. A., Joung, J. K., Kohn, D. B., Ozawa, K., & Sadelain, M. (2018) · Science · doi:10.1126/science.aan4672 · Cited by 1,410
“In addition, the regulation of an introduced or corrected gene will be controlled by the endogenous promoter, resulting in more physiologic and appropriately regulated gene expression (63).”
Self-citation‘Off-the-shelf’ allogeneic CAR T cells: development and challenges
Depil, S., Duchateau, P. N., Grupp, S. A., Mufti, G. J., & Poirot, L. (2020) · Nature Reviews Drug Discovery · doi:10.1038/s41573-019-0051-2 · Cited by 1,266
Identification of preexisting adaptive immunity to Cas9 proteins in humans
Charlesworth, C. T., Deshpande, P. S., Dever, D. P., Camarena, J., Lemgart, V. T., Cromer, M. K., Vakulskas, C. A., Collingwood, M. A., Zhang, L., Bode, N. M., et al. (2019) · Nature Medicine · doi:10.1038/s41591-018-0326-x · Cited by 1,093
Mechanisms of resistance to CAR T cell therapy
Shah, N. N., & Fry, T. J. (2019) · Nature Reviews Clinical Oncology · doi:10.1038/s41571-019-0184-6 · Cited by 999
Clinical implications of T cell exhaustion for cancer immunotherapy
Chow, A., Perica, K., Klebanoff, C. A., & Wolchok, J. D. (2022) · Nature Reviews Clinical Oncology · doi:10.1038/s41571-022-00689-z · Cited by 1,044
c-Jun overexpression in CAR T cells induces exhaustion resistance
Lynn, R. C., Weber, E. W., Sotillo, E., Gennert, D., Xu, P., Good, Z., Anbunathan, H., Lattin, J., Jones, R., Tieu, V., et al. (2019) · Nature · doi:10.1038/s41586-019-1805-z · Cited by 891
Reprogramming human T cell function and specificity with non-viral genome targeting
Roth, T. L., Puig-Saus, C., Yu, R., Shifrut, E., Carnevale, J., Li, P. J., Hiatt, J., Saco, J., Krystofinski, P., Li, H., et al. (2018) · Nature · doi:10.1038/s41586-018-0326-5 · Cited by 839
Therapeutic T cell engineering
Sadelain, M., Rivière, I., & Riddell, S. R. (2017) · Nature · doi:10.1038/nature22395 · Cited by 906
Self-citationMartinez, M. C., & Moon, E. K. (2019) · Frontiers in Immunology · doi:10.3389/fimmu.2019.00128 · Cited by 865
“A study with a CD19/28ζ CAR used CRISPR to insert the CAR under the control of the TCR promoter (at the TRAC locus), while simultaneously knocking out the TCR via insertion of the CAR gene (182).”
MethodRecent advances and discoveries in the mechanisms and functions of CAR T cells
Larson, R. C., & Maus, M. V. (2021) · Nature Reviews Cancer · doi:10.1038/s41568-020-00323-z · Cited by 852
Vakulskas, C. A., Dever, D. P., Rettig, G. R., Turk, R., Jacobi, A. M., Collingwood, M. A., Bode, N. M., McNeill, M. S., Yan, S., Camarena, J., et al. (2018) · Nature Medicine · doi:10.1038/s41591-018-0137-0 · Cited by 782
Tools for translation: non-viral materials for therapeutic mRNA delivery
Hajj, K. A., & Whitehead, K. A. (2017) · Nature Reviews Materials · doi:10.1038/natrevmats.2017.56 · Cited by 799
Treg cell-based therapies: challenges and perspectives
Raffin, C., Vo, L. T., & Bluestone, J. A. (2019) · Nature reviews. Immunology · doi:10.1038/s41577-019-0232-6 · Cited by 746
CAR immune cells: design principles, resistance and the next generation
Labanieh, L., & Mackall, C. L. (2023) · Nature · doi:10.1038/s41586-023-05707-3 · Cited by 695
CAR T cell trogocytosis and cooperative killing regulate tumour antigen escape
Hamieh, M., Dobrin, A., Cabriolu, A., van der Stegen, S. J. C., Giavridis, T., Mansilla‐Soto, J., Eyquem, J., Zhao, Z., Whitlock, B. M., Miele, M. M., et al. (2019) · Nature · doi:10.1038/s41586-019-1054-1 · Cited by 688
Self-citationClinical lessons learned from the first leg of the CAR T cell journey
Majzner, R. G., & Mackall, C. L. (2019) · Nature Medicine · doi:10.1038/s41591-019-0564-6 · Cited by 636
Targeting the epigenetic regulation of antitumour immunity
Hogg, S. J., Beavis, P. A., Dawson, M. A., & Johnstone, R. W. (2020) · Nature Reviews Drug Discovery · doi:10.1038/s41573-020-0077-5 · Cited by 665
Engineering CAR-T Cells for Next-Generation Cancer Therapy
Hong, M., Clubb, J. D., & Chen, Y. Y. (2020) · Cancer Cell · doi:10.1016/j.ccell.2020.07.005 · Cited by 659
An introduction to chimeric antigen receptor (CAR) T‐cell immunotherapy for human cancer
Feins, S., Kong, W., Williams, E. F., Milone, M. C., & Fraietta, J. A. (2019) · American Journal of Hematology · doi:10.1002/ajh.25418 · Cited by 614
“guided integration of antigen receptor transgenes into specific loci using viral and non-viral methods.(48,49) Compositions of CD4 and CD8 T cell subsets differ in cancer”
MethodKagoya, Y., Tanaka, S., Guo, T., Anczurowski, M., Wang, C.-H., Saso, K., Butler, M. O., Minden, M. D., & Hirano, N. (2018) · Nature Medicine · doi:10.1038/nm.4478 · Cited by 654
Transient rest restores functionality in exhausted CAR-T cells through epigenetic remodeling
Weber, E. W., Parker, K. R., Sotillo, E., Lynn, R. C., Anbunathan, H., Lattin, J., Good, Z., Belk, J. A., Dániel, B., Klysz, D. D., et al. (2021) · Science · doi:10.1126/science.aba1786 · Cited by 646
“T cell exhaustion has been implicated as an important factor limiting the efficacy of CAR-T cells against cancer (4-6) and can be driven by excessive CAR signaling as a result of high antigen burden or tonic signaling induced by antigen-independent clustering of the CAR receptor (7, 8).”
Past, present, and future of CRISPR genome editing technologies
Pacesa, M., Pelea, O., & Jinek, M. (2024) · Cell · doi:10.1016/j.cell.2024.01.042 · Cited by 609
“276 CRISPR-directed insertion of the CAR construct at the T cell receptor a constant (TRAC) locus using HDR avoids graft-versus-host disease, 277 whereas knockouts of PDCD1, Regnase-1, and TGFBR2 genes are designed to reduce CAR-T cell exhaustion and increase persistence.”
Enhancing CAR T cell persistence through ICOS and 4-1BB costimulation
Guedan, S., Posey, A. D., Shaw, C. E., Wing, A., Da, T., Patel, P. R., McGettigan, S. E., Casado‐Medrano, V., Kawalekar, O. U., Uribe‐Herranz, M., et al. (2018) · JCI Insight · doi:10.1172/jci.insight.96976 · Cited by 595
“use of the 4-1BB ICD (25), optimization of the spacer length (50), targeting of a CAR to the T cell receptor α constant TRAC locus (52), or use of self-inactivating LVs instead of gammaretroviral vectors (51).”
MethodKilling Mechanisms of Chimeric Antigen Receptor (CAR) T Cells
Benmebarek, M.-R., Karches, C. H., Cadilha, B. L., Lesch, S., Endres, S., & Kobold, S. (2019) · International Journal of Molecular Sciences · doi:10.3390/ijms20061283 · Cited by 550
“Signaling domain configurations, spacer length [117], and gene transfer approaches that could impact CAR expression levels on the T cell surface must be considered [118,119].”
BackgroundThe tyrosine kinase inhibitor dasatinib acts as a pharmacologic on/off switch for CAR T cells
Mestermann, K., Giavridis, T., Weber, J., Rydzek, J., Frenz, S., Nerreter, T., Mades, A., Sadelain, M., Einsele, H., & Hudecek, M. (2019) · Science Translational Medicine · doi:10.1126/scitranslmed.aau5907 · Cited by 531
Self-citationCalibration of CAR activation potential directs alternative T cell fates and therapeutic potency
Feucht, J., Sun, J., Eyquem, J., Ho, Y.-J., Zhao, Z., Leibold, J., Dobrin, A., Cabriolu, A., Hamieh, M., & Sadelain, M. (2019) · Nature Medicine · doi:10.1038/s41591-018-0290-5 · Cited by 547
Self-citationCRISPR-Based Therapeutic Genome Editing: Strategies and In Vivo Delivery by AAV Vectors
Wang, D., Zhang, F., & Gao, G. (2020) · Cell · doi:10.1016/j.cell.2020.03.023 · Cited by 534
“Using CRISPR to achieve targeted integration of the CAR by HDR can achieve concomitant disruption of the T-cell receptor a constant (TRAC) locus, resulting in engineered CAR-T cells that may be more amenable to allogeneic infusion (Eyquem et al., 2017).”
Chimeric antigen receptor T-cell therapies for lymphoma
Brudno, J. N., & Kochenderfer, J. N. (2017) · Nature Reviews Clinical Oncology · doi:10.1038/nrclinonc.2017.128 · Cited by 522
Salter, A. I., Ivey, R. G., Kennedy, J. J., Voillet, V., Rajan, A., Alderman, E. J., Voytovich, U. J., Lin, C., Sommermeyer, D., Liu, L., et al. (2018) · Science Signaling · doi:10.1126/scisignal.aat6753 · Cited by 503
mRNA-based cancer therapeutics
Liu, C., Shi, Q., Huang, X., Koo, S., Kong, N., & Tao, W. (2023) · Nature Reviews Cancer · doi:10.1038/s41568-023-00586-2 · Cited by 500
Next-generation regulatory T cell therapy
Ferreira, L. M. R., Müller, Y. D., Bluestone, J. A., & Tang, Q. (2019) · Nature Reviews Drug Discovery · doi:10.1038/s41573-019-0041-4 · Cited by 491
Yarnall, M. T. N., Ioannidi, E. I., Schmitt-Ulms, C., Krajeski, R. N., Lim, J., Villiger, L., Zhou, W., Jiang, K., Garushyants, S. K., Roberts, N., et al. (2022) · Nature Biotechnology · doi:10.1038/s41587-022-01527-4 · Cited by 503
Beyond immune checkpoint blockade: emerging immunological strategies
Kubli, S. P., Berger, T., Araújo, D. V., Siu, L. L., & Mak, T. W. (2021) · Nature Reviews Drug Discovery · doi:10.1038/s41573-021-00155-y · Cited by 460
Engineering and Design of Chimeric Antigen Receptors
Guedan, S., Calderón, H., Posey, A. D., & Maus, M. V. (2018) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2018.12.009 · Cited by 469
The Emerging Landscape of Immune Cell Therapies
Weber, E. W., Maus, M. V., & Mackall, C. L. (2020) · Cell · doi:10.1016/j.cell.2020.03.001 · Cited by 450
“Insertion of the CAR transgene into the TRAC locus and endogenous control of CAR expression prevented exhaustion in pre-clinical leukemia models (Eyquem et al., 2017).”
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Bashor, C. J., Hilton, I. B., Bandukwala, H. S., Smith, D. M., & Omid Veiseh, A. (2022) · Nature Reviews Drug Discovery · doi:10.1038/s41573-022-00476-6 · Cited by 472
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Ma, L., Dichwalkar, T., Chang, J. Y. H., Cossette, B. J., Garafola, D., Zhang, A. Q., Fichter, M., Wang, C., Liang, S., Silva, M., et al. (2019) · Science · doi:10.1126/science.aav8692 · Cited by 439
Current applications and future perspective of CRISPR/Cas9 gene editing in cancer
Wang, S.-W., Gao, C., Zheng, Y.-M., Yi, L., Lu, J.-C., Huang, X.-Y., Cai, J.-B., Zhang, P.-F., Cui, Y.-H., & Ke, A.-W. (2022) · Molecular Cancer · doi:10.1186/s12943-022-01518-8 · Cited by 478
“Indeed, using the CRISPR system to direct CAR to the TRAC locus can avoid tonic CAR signal transduction and establish effective CAR internalization and reexpression after cells are exposed to antigen, effectively delaying differentiation and exhaustion of effector T cells [191].”
MethodProgramming CAR-T cells to kill cancer
Labanieh, L., Majzner, R. G., & Mackall, C. L. (2018) · Nature Biomedical Engineering · doi:10.1038/s41551-018-0235-9 · Cited by 450
TCR-engineered T cell therapy in solid tumors: State of the art and perspectives
Baulu, E., Gardet, C., Chuvin, N., & Depil, S. (2023) · Science Advances · doi:10.1126/sciadv.adf3700 · Cited by 474
CRISPR in cancer biology and therapy
Katti, A., Diaz, B. J., Caragine, C. M., Sanjana, N. E., & Dow, L. E. (2022) · Nature reviews. Cancer · doi:10.1038/s41568-022-00441-w · Cited by 451
Current understanding of osteoarthritis pathogenesis and relevant new approaches
Tong, L., Yu, H., Huang, X., Shen, J., Xiao, G., Chen, L., Wang, H., Xing, L., & Chen, D. (2022) · Bone Research · doi:10.1038/s41413-022-00226-9 · Cited by 437
“A particularly inspiring direction is the development of Cas9 as a therapeutic molecule for curing hereditary diseases.(160,161) Delivery tools for gene-editing molecules include adeno-associated viral vectors, lentiviral vectors, lipid nanoparticles, and other nonviral vectors.”
BackgroundInhibitory CD161 receptor identified in glioma-infiltrating T cells by single-cell analysis
Mathewson, N. D., Ashenberg, O., Tirosh, I., Gritsch, S., Perez, E. M., Marx, S., Jerby‐Arnon, L., Chanoch-Myers, R., Hara, T., Richman, A. R., et al. (2021) · Cell · doi:10.1016/j.cell.2021.01.022 · Cited by 415
“CD4 crRNA: (50-GGCAAGGCCACAATGAACCG-30) This paper; IDT N/A TRAC crRNA: (50-TCAGGGTTCTGGATATCTGT-30) (Eyquem et al., 2017); IDT N/A KLRB1 region#1 crRNA: (50-AATTAAAGCCACTTACCCCG-30) This paper; IDT N/A KLRB1 region#2 crRNA: (50-TTACCCCGAGGAAGAGATGA-30) This paper; IDT N/A”
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RNA Drugs and RNA Targets for Small Molecules: Principles, Progress, and Challenges
Yu, A., Choi, Y. H., & Tu, M. (2020) · Pharmacological Reviews · doi:10.1124/pr.120.019554 · Cited by 421
“…of new therapies for the treatment of human diseases, including monogenetic disorders, infection, and cancer (Xue et al., 2014; Dever et al., 2016; Long et al., 2016; Nelson et al., 2016; Tabebordbar et al., 2016; Eyquem et al., 2017; Zhang et al., 2017; Georgiadis et al., 2018; Xu et al., 2019b).”
BackgroundAn “off-the-shelf” fratricide-resistant CAR-T for the treatment of T cell hematologic malignancies
Cooper, M. L., Choi, J., Staser, K. W., Ritchey, J., Devenport, J. M., Eckardt, K., Rettig, M. P., Wang, B., Eissenberg, L. G., Ghobadi, A., et al. (2018) · Leukemia · doi:10.1038/s41375-018-0065-5 · Cited by 396
“Furthermore, insertion of the CAR directly into the TRAC locus, as recently reported [40, 41], or, potentially, the CD7 locus, would further mitigate the risk”
BackgroundEmerging Cellular Therapies for Cancer
Guedan, S., Ruella, M., & June, C. H. (2018) · Annual Review of Immunology · doi:10.1146/annurev-immunol-042718-041407 · Cited by 393
CXCR1- or CXCR2-modified CAR T cells co-opt IL-8 for maximal antitumor efficacy in solid tumors
Jin, L., Tao, H., Karachi, A., Long, Y., Hou, A. Y., Meng, N., Dyson, K. A., Grippin, A., Deleyrolle, L. P., Zhang, W., et al. (2019) · Nature Communications · doi:10.1038/s41467-019-11869-4 · Cited by 392
Choi, J., Lim, J., Shin, M., Paek, S., & Choi, J.-W. (2020) · Nano Letters · doi:10.1021/acs.nanolett.0c04303 · Cited by 386
CAR T Cell Therapy for Solid Tumors: Bright Future or Dark Reality?
Wagner, J., Wickman, E., DeRenzo, C. C., & Gottschalk, S. M. (2020) · Molecular Therapy · doi:10.1016/j.ymthe.2020.09.015 · Cited by 395
“This resulted in consistent CAR expression, reduced baseline signaling, and improved antitumor activity compared to CD19-CAR T cells generated by viral transduction.(125) The clinical experience with CAR T cells generated with non-viral methods is so far limited,(78) and most ongoing studies are focused on targeting h…”
BackgroundCAR-T design: Elements and their synergistic function
Jayaraman, J., Mellody, M., Hou, A. J., Desai, R. P., Fung, A., Pham, A. H. T., Chen, Y. Y., & Zhao, W. (2020) · EBioMedicine · doi:10.1016/j.ebiom.2020.102931 · Cited by 403
“reduced tonic signaling and improved in vivo anti-tumor efficacy [35].”
BackgroundChimeric antigen receptor natural killer (CAR-NK) cell design and engineering for cancer therapy
Gong, Y., Klein Wolterink, R. G. J., Wang, J., Bos, G. M. J., & Germeraad, W. T. V. (2021) · Journal of Hematology & Oncology · doi:10.1186/s13045-021-01083-5 · Cited by 379
“TRAC-CAR-T outperformed both conventional retroviral transduced CAR T cells (RVCAR) as well as T cells with the CAR inserted in the β2M locus (β2M-CAR) in an in vivo mouse model, based on tumor eradication and expression of exhaustion markers, including PD1, TIM3 and LAG3 [40].”
BackgroundMethodInfluentialNon-viral, specifically targeted CAR-T cells achieve high safety and efficacy in B-NHL
Zhang, J., Hu, Y., Yang, J., Li, W., Zhang, M., Wang, Q., Zhang, L., Wei, G., Tian, Y., Zhao, K., et al. (2022) · Nature · doi:10.1038/s41586-022-05140-y · Cited by 397
Seki, A., & Rutz, S. (2018) · The Journal of Experimental Medicine · doi:10.1084/jem.20171626 · Cited by 369
CAR T cell killing requires the IFNγR pathway in solid but not liquid tumours
Larson, R. C., Kann, M. C., Bailey, S. R., Haradhvala, N. J., Llopis, P. D. L. M. M., Bouffard, A. A., Scarfò, I., Leick, M. B., Grauwet, K., Berger, T. R., et al. (2022) · Nature · doi:10.1038/s41586-022-04585-5 · Cited by 365
Engineering the next-generation of CAR T-cells with CRISPR-Cas9 gene editing
Dimitri, A. J., Herbst, F., & Fraietta, J. A. (2022) · Molecular Cancer · doi:10.1186/s12943-022-01559-z · Cited by 379
Xu, X., Sun, Q., Liang, X., Chen, Z., Zhang, X., Zhou, X., Li, M., Tu, H., Liu, Y., Tu, S., et al. (2019) · Frontiers in Immunology · doi:10.3389/fimmu.2019.02664 · Cited by 360
“(48) reported that targeting a CD19-specific CAR coding sequence to the T-cell receptor (TCR) locus, placing it under the control of endogenous regulatory elements, not only led to uniform CAR expression in human peripheral blood Tcells but also promoted T-cell potency.”
BackgroundBlocking expression of inhibitory receptor NKG2A overcomes tumor resistance to NK cells
Kamiya, T., Seow, S. V., Wong, D., Robinson, M., & Campana, D. (2019) · Journal of Clinical Investigation · doi:10.1172/jci123955 · Cited by 345
“Downregulation of NKG2A might have also been achieved by using modern gene-editing methodologies based on mega-nucleases, TALEN, or CRISPR/Cas9, which have already been applied to genetically modify T lymphocytes (66–69).”
Principles and therapeutic applications of adaptive immunity
Chi, H., Pepper, M., & Thomas, P. G. (2024) · Cell · doi:10.1016/j.cell.2024.03.037 · Cited by 394
Coexpression profile of leukemic stem cell markers for combinatorial targeted therapy in AML
Haubner, S., Perna, F., Köhnke, T., Schmidt, C., Berman, S., Augsberger, C., Schnorfeil, F. M., Krupka, C., Lichtenegger, F. S., Liu, X., et al. (2018) · Leukemia · doi:10.1038/s41375-018-0180-3 · Cited by 351
Self-citationSystematic Immunotherapy Target Discovery Using Genome-Scale In Vivo CRISPR Screens in CD8 T Cells
Dong, M. B., Wang, G., Chow, R. D., Ye, L., Zhu, L., Dai, X., Park, J. J., Kim, H. R., Errami, Y., Guzman, C. D., et al. (2019) · Cell · doi:10.1016/j.cell.2019.07.044 · Cited by 330
“CRISPR technology has provided a unique opportunity for direct manipulation of the T cell genome (Eyquem et al., 2017), and its scalability enabled genetic screens in T cells which to date have been performed in vitro (Henriksson et al.”
CRISPR/Cas9 for cancer research and therapy
Zhan, T., Rindtorff, N., Betge, J., Ebert, M., & Boutros, M. (2018) · Seminars in Cancer Biology · doi:10.1016/j.semcancer.2018.04.001 · Cited by 338
T cell receptor‐based cancer immunotherapy: Emerging efficacy and pathways of resistance
Chandran, S. S., & Klebanoff, C. A. (2019) · Immunological Reviews · doi:10.1111/imr.12772 · Cited by 335
Epigenetic regulation of T cell exhaustion
Belk, J. A., Dániel, B., & Satpathy, A. T. (2022) · Nature Immunology · doi:10.1038/s41590-022-01224-z · Cited by 346
Scalable Manufacturing of CAR T Cells for Cancer Immunotherapy
Abou-el-Enein, M., Elsallab, M., Feldman, S. A., Fesnak, A. D., Heslop, H. E., Marks, P., Till, B. G., Bauer, G., & Savoldo, B. (2021) · Blood Cancer Discovery · doi:10.1158/2643-3230.bcd-21-0084 · Cited by 321
“This strategy enables the disruption of the TCR by targeting the T-cell receptor alpha chain constant (TRAC) locus while simultaneously integrating the CAR construct into the genome (150).”
Rapid manufacturing of non-activated potent CAR T cells
Ghassemi, S., Durgin, J. S., Nuñez-Cruz, S., Patel, J., Leferovich, J. M., Pinzone, M. R., Shen, F., Cummins, K. D., Plesa, G., Cantu, V. A., et al. (2022) · Nature Biomedical Engineering · doi:10.1038/s41551-021-00842-6 · Cited by 316
CAR-T cell manufacturing: Major process parameters and next-generation strategies
Ceja, M. A., Khericha, M., Harris, C. M., Puig-Saus, C., & Chen, Y. Y. (2023) · The Journal of Experimental Medicine · doi:10.1084/jem.20230903 · Cited by 349
“This strategy commandeers the endogenous gene regulatory mechanisms that dynamically control TCR expression levels in response to T-cell activation states, with data suggesting that the resulting CAR-T cells may be more efficacious than virally integrated CAR-T cells (Eyquem et al., 2017).”
BackgroundCAR T cells: Continuation in a revolution of immunotherapy
Singh, A. K., & McGuirk, J. P. (2020) · The Lancet Oncology · doi:10.1016/s1470-2045(19)30823-x · Cited by 303
Zhao, Y., Chen, J., Andreatta, M., Feng, B., Xie, Y., Wenes, M., Wang, Y., Gao, M., Hu, X., Romero, P. J., et al. (2024) · Nature Biotechnology · doi:10.1038/s41587-023-02060-8 · Cited by 309
“The CD19 hCAR comprised an scFv specific for human CD19 preceded by a CD8α leader peptide and followed by CD28 hinge–transmembrane–intracellular regions and a CD3ζ intracellular domain, as described in previous publications 34 .”
Recent Advances in CRISPR/Cas9 Delivery Strategies
Yip, B. (2020) · Biomolecules · doi:10.3390/biom10060839 · Cited by 292
Current status and perspectives of patient-derived xenograft models in cancer research
Lai, Y., Wei, X., Lin, S., Qin, L., Cheng, L., & Li, P. (2017) · Journal of Hematology & Oncology · doi:10.1186/s13045-017-0470-7 · Cited by 308
“The in vivo model with NSG mice was critical to demonstrate that targeting an anti-CD19 CAR to the TRAC locus with CRISPR/Cas9 enhances tumor rejection, a strategy averting antigen-stimulated differentiation and exhaustion [124].”
BackgroundChoi, B. D., Yu, X., Castaño, A. P., Darr, H., Henderson, D. B., Bouffard, A. A., Larson, R. C., Scarfò, I., Bailey, S. R., Gerhard, G. M., et al. (2019) · Journal for ImmunoTherapy of Cancer · doi:10.1186/s40425-019-0806-7 · Cited by 293
“An example of this is a prior report of a CD19 CAR construct delivered directly into the TRAC locus, which also placed the transgene under control of an endogenous promoter [7].”
BackgroundNon-viral precision T cell receptor replacement for personalized cell therapy
Foy, S. P., Jacoby, K., Bota, D. A., Hunter, T., Pan, Z., Stawiski, E. W., Ma, Y., Lu, W., Peng, S., Wang, C. L., et al. (2022) · Nature · doi:10.1038/s41586-022-05531-1 · Cited by 301
Gomes‐Silva, D., Mukherjee, M., Srinivasan, M., Krenciute, G., Dakhova, O., Zheng, Y., Cabral, J. M. S., Rooney, C. M., Orange, J. S., Brenner, M. K., et al. (2017) · Cell Reports · doi:10.1016/j.celrep.2017.09.015 · Cited by 293
The delivery challenge: fulfilling the promise of therapeutic genome editing
van Haasteren, J., Li, J., Scheideler, O. J., Murthy, N., & Schaffer, D. V. (2020) · Nature Biotechnology · doi:10.1038/s41587-020-0565-5 · Cited by 285
CHANGE-seq reveals genetic and epigenetic effects on CRISPR–Cas9 genome-wide activity
Lazzarotto, C. R., Malinin, N. L., Li, Y., Zhang, R., Yang, Y., Lee, G., Cowley, E., He, Y., Lan, X., Jividen, K., et al. (2020) · Nature Biotechnology · doi:10.1038/s41587-020-0555-7 · Cited by 289
Next-generation stem cells — ushering in a new era of cell-based therapies
Kimbrel, E. A., & Lanza, R. (2020) · Nature Reviews Drug Discovery · doi:10.1038/s41573-020-0064-x · Cited by 295
Nidhi, S., Anand, U., Olekšák, P., Tripathi, P., Lal, J. A., Thomas, G., Kuča, K., & Tripathi, V. S. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms22073327 · Cited by 310
Low-Dose Radiation Conditioning Enables CAR T Cells to Mitigate Antigen Escape
DeSelm, C. J., Palomba, M. L., Yahalom, J., Hamieh, M., Eyquem, J., Rajasekhar, V. K., & Sadelain, M. (2018) · Molecular Therapy · doi:10.1016/j.ymthe.2018.09.008 · Cited by 280
“For TCR knockout, we used a gRNA that targets a sequence in the first exon of the constant chain of the TCRa gene (TRAC) that is required for the TCRa and b assembly and addressing it to the cellsurface, as described previously.(41) TRAIL was performed using synthetic modified gRNA kits (Synthego).”
MethodSelf-citationReversible ON- and OFF-switch chimeric antigen receptors controlled by lenalidomide
Jan, M., Scarfò, I., Larson, R. C., Walker, A., Schmidts, A., Guirguis, A. A., Gasser, J. A., Slabicki, M. M., Bouffard, A. A., Castaño, A. P., et al. (2021) · Science Translational Medicine · doi:10.1126/scitranslmed.abb6295 · Cited by 284
“In future applications, if necessary, effects of degron or dimerization domains on receptor function could be mitigated by altering gene dosage or per-molecule signaling strength along established lines ( 34 , 36 , 42 , 43 ).”
Du, B., Qin, J., Lin, B., Zhang, J., Li, D., & Liu, M. (2025) · Cancer Cell · doi:10.1016/j.ccell.2025.03.019 · Cited by 285
Phase I study of CAR-T cells with PD-1 and TCR disruption in mesothelin-positive solid tumors
Wang, Z., Li, N., Feng, K., Chen, M., Zhang, Y., Liu, Y., Yang, Q., Nie, J., Tang, N., Zhang, X., et al. (2021) · Cellular and Molecular Immunology · doi:10.1038/s41423-021-00749-x · Cited by 281
Sheih, A., Voillet, V., Hanafi, L., DeBerg, H. A., Yajima, M., Hawkins, R. M., Gersuk, V. H., Riddell, S. R., Maloney, D. G., Wohlfahrt, M. E., et al. (2020) · Nature Communications · doi:10.1038/s41467-019-13880-1 · Cited by 268
Challenges and new technologies in adoptive cell therapy
Zhang, P., Zhang, G., & Wan, X. (2023) · Journal of Hematology & Oncology · doi:10.1186/s13045-023-01492-8 · Cited by 291
“TCR site destruction and CAR coding gene insertion by CRISPR/Cas9 has been reported to result in potentially universal CAR-T cells that effectively reduced tonic signaling and delayed exhaustion [172, 173].”
BackgroundMethodNanotechnology Strategies To Advance Outcomes in Clinical Cancer Care
Hartshorn, C. M., Bradbury, M. S., Lanza, G. M., Nel, A. E., Rao, J., Wang, A. Z., Wiesner, U. B., Yang, L. P. H., & Grodzinski, P. A. (2017) · ACS Nano · doi:10.1021/acsnano.7b05108 · Cited by 268
Wittsten, A. H., Yang, S., Park, M., Garcia, J. M., Alavi, J., Perry, N., Montgomery, G., Liu, B., & Roybal, K. T. (2021) · Science Translational Medicine · doi:10.1126/scitranslmed.abd8836 · Cited by 262
Boyiadzis, M. M., Dhodapkar, M. V., Brentjens, R. J., Kochenderfer, J. N., Neelapu, S. S., Maus, M. V., Porter, D. L., Maloney, D. G., Grupp, S. A., Mackall, C. L., et al. (2018) · Journal for ImmunoTherapy of Cancer · doi:10.1186/s40425-018-0460-5 · Cited by 255
Zhang, H., Qin, C., An, C., Zheng, X., Wen, S., Chen, W., Liu, X., Lv, Z., Yang, P., Xu, W., et al. (2021) · Molecular Cancer · doi:10.1186/s12943-021-01431-6 · Cited by 264
Watanabe, K., Kuramitsu, S., Posey, A. D., & June, C. H. (2018) · Frontiers in Immunology · doi:10.3389/fimmu.2018.02486 · Cited by 248
“This difference in sustained lysis kinetics may be explained by CAR downmodulation after stimulation upon the antigen recognition, which can be ameliorated through TCR-based expression of CAR (41).”
BackgroundKuhn, N. F., Purdon, T. J., van Leeuwen, D. G., Lopez, A. V., Curran, K. J., Daniyan, A. F., & Brentjens, R. J. (2019) · Cancer Cell · doi:10.1016/j.ccell.2019.02.006 · Cited by 253
“by controlling CAR expression at a physiological level through targeted genomic integration of the CAR transgene (Eyquem et al., 2017), or by expressing a second transgene in addition to the CAR in T cells to enhance the antitumor efficacy of CAR T cells (Pegram et al.”
BackgroundGenetically engineered T cells for cancer immunotherapy
Li, D., Li, X., Zhou, W.-L., Huang, Y., Liang, X., Jiang, L., Yang, X., Sun, J., Li, Z., Han, W., et al. (2019) · Signal Transduction and Targeted Therapy · doi:10.1038/s41392-019-0070-9 · Cited by 251
Cellular networks controlling T cell persistence in adoptive cell therapy
Chan, J. D., Lai, J., Slaney, C. Y., Kallies, A., Beavis, P. A., & Darcy, P. K. (2021) · Nature reviews. Immunology · doi:10.1038/s41577-021-00539-6 · Cited by 253
Metadata
- Authors
- Eyquem, J., Mansilla-Soto, J., Giavridis, T., van der Stegen, S. J. C., Hamieh, M., Cunanan, K. M., Odak, A., Gönen, M., & Sadelain, M.
- Affiliations
- Memorial Sloan Kettering Cancer Center (US)
- Corresponding author
- Sadelain, Michel
- ORCID
- Venue
- Nature
- Published
- 2017-02-22
- Volume
- 543
- Issue
- 7643
- Pages
- 113-117
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC5558614
- OpenAlex
- W2589417329
- Semantic Scholar
- 47af46911569825ab86ac946a5c9cd502504aa3f
- Cited by
- 1,831
- References
- 33
- Citations by year
- 2026: 191 · 2025: 175 · 2024: 206 · 2023: 201 · 2022: 224 · 2021: 238 · 2020: 190 · 2019: 186 · 2018: 151 · 2017: 68
- FWCI
- 71.74
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Funders
- National Cancer Institute (P30CA008748)
- Topics
- Health Sciences › Medicine › Oncology › CAR-T cell therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Keywords
- chimeric antigen receptor, CRISPR/Cas9 genome editing, CD19, chimeric antigen receptor T cells, T cell exhaustion, T cell differentiation, acute lymphoblastic leukemia, tumor rejection, targeted gene integration
- MeSH terms
- Gene Editing, Animals, Cell Differentiation, Disease Models, Animal, Gene Expression Regulation, Humans, Immunotherapy, Lymphocyte Activation, Male, Promoter Regions, Genetic, Receptors, Antigen, T-Cell, T-Lymphocytes, Receptors, Antigen, T-Cell, alpha-beta, Antigens, CD19, Mice, Precursor Cell Lymphoblastic Leukemia-Lymphoma, Genetic Loci, Translational Research, Biomedical, CRISPR-Cas Systems
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