TY - JOUR
T1 - Synergistic T cell signaling by 41BB and CD28 is optimally achieved by membrane proximal positioning within parallel chimeric antigen receptors
AU - Muliaditan, Tamara
AU - Halim, Leena
AU - Whilding, Lynsey
AU - Draper, Benjamin
AU - Achkova, Daniela
AU - Kausar, Fahima
AU - Glover, Maya
AU - Bechman, Natasha
AU - Arulappu, Appitha
AU - Sanchez, Jenifer
AU - Flaherty, Katie R.
AU - Obajdin, Jana
AU - Grigoriadis, Kristiana
AU - Antoine, Pierre
AU - Larcombe-Young, Daniel
AU - Hull, Caroline
AU - Buus, Richard
AU - Gordon, Peter
AU - Grigoriadis, Anita
AU - Davies, David
AU - Schurich, Anna
AU - Maher, John
N1 - Funding Information:
We thank M. van Schalkwyk and A. Adami for critical review of the manuscript and scientific colleagues for provision of materials. This work was supported by Leucid Bio ; Cancer Research UK (CRUK) grant number A21623 ; the British Lung Foundation ; the Experimental Cancer Medicine Centre at King’s College London ; the King’s Health Partners/King’s College London Cancer Research UK Cancer Centre ; Kings Health Partners MRC Confidence in Concepts ; a Joint Research Committee Kings Medical Research Fund PhD studentship; a Wellcome Trust PhD studentship grant number 108874/B/15/Z ; the National Institute for Health Research (NIHR) Biomedical Research Centre, based at Guy’s and St Thomas’ NHS Foundation Trust and King’s College London; a CRUK-KHP fast-track seed award; and a MRC-DTP PhD studentship. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health.
Funding Information:
We thank M. van Schalkwyk and A. Adami for critical review of the manuscript and scientific colleagues for provision of materials. This work was supported by Leucid Bio; Cancer Research UK (CRUK) grant number A21623; the British Lung Foundation; the Experimental Cancer Medicine Centre at King's College London; the King's Health Partners/King's College London Cancer Research UK Cancer Centre; Kings Health Partners MRC Confidence in Concepts; a Joint Research Committee Kings Medical Research Fund PhD studentship; a Wellcome Trust PhD studentship grant number 108874/B/15/Z; the National Institute for Health Research (NIHR) Biomedical Research Centre, based at Guy's and St Thomas? NHS Foundation Trust and King's College London; a CRUK-KHP fast-track seed award; and a MRC-DTP PhD studentship. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health. Conceptualization, J.M.; experimental work, T.M. L.H. L.M.W. B.D. D.Y.A. F.K. M.G. N.B. A.A. J.S. K.R.F. P.A. D.L.-Y. C.M.H. R.B. P.G. D.M.D. and A.S.; bioinformatic analysis, T.M. K.G. J.O. and A.G.; methodology and data analysis, all authors; writing ? original draft, J.M. A.S. and T.M.; writing ? review & editing, all authors. J.M. is CSO, scientific founder, and shareholder of Leucid Bio. T.M. F.K. M.G. and A.A. are employees of Leucid Bio. L.H. and B.D. undertook PhD studentships funded by Leucid Bio. L.M.W. D.M.D. C.H. and D.L.-Y. have acted as consultants for Leucid Bio, and D.M.D. is currently an employee of Leucid Bio. J.M. D.Y.A. L.M.W. B.D. T.M. F.K. L.H. and M.G. are co-inventors on patent filings in relation to pCAR technology. B.D. and D.Y.A. are shareholders of Autolus Therapeutics. D.Y.A. is an employee of Autolus Therapeutics. The other authors declare no competing interests.
Publisher Copyright:
© 2021 The Author(s)
PY - 2021/12/21
Y1 - 2021/12/21
N2 - Second generation (2G) chimeric antigen receptors (CARs) contain a CD28 or 41BB co-stimulatory endodomain and elicit remarkable efficacy in hematological malignancies. Third generation (3G) CARs extend this linear blueprint by fusing both co-stimulatory units in series. However, clinical impact has been muted despite compelling evidence that co-signaling by CD28 and 41BB can powerfully amplify natural immune responses. We postulate that effective dual co-stimulation requires juxta-membrane positioning of endodomain components within separate synthetic receptors. Consequently, we designed parallel (p)CARs in which a 2G (CD28+CD3ζ) CAR is co-expressed with a 41BB-containing chimeric co-stimulatory receptor. We demonstrate that the pCAR platform optimally harnesses synergistic and tumor-dependent co-stimulation to resist T cell exhaustion and senescence, sustaining proliferation, cytokine release, cytokine signaling, and metabolic fitness upon repeated stimulation. When engineered using targeting moieties of diverse composition, affinity, and specificity, pCAR T cells consistently elicit superior anti-tumor activity compared with T cells that express traditional linear CARs.
AB - Second generation (2G) chimeric antigen receptors (CARs) contain a CD28 or 41BB co-stimulatory endodomain and elicit remarkable efficacy in hematological malignancies. Third generation (3G) CARs extend this linear blueprint by fusing both co-stimulatory units in series. However, clinical impact has been muted despite compelling evidence that co-signaling by CD28 and 41BB can powerfully amplify natural immune responses. We postulate that effective dual co-stimulation requires juxta-membrane positioning of endodomain components within separate synthetic receptors. Consequently, we designed parallel (p)CARs in which a 2G (CD28+CD3ζ) CAR is co-expressed with a 41BB-containing chimeric co-stimulatory receptor. We demonstrate that the pCAR platform optimally harnesses synergistic and tumor-dependent co-stimulation to resist T cell exhaustion and senescence, sustaining proliferation, cytokine release, cytokine signaling, and metabolic fitness upon repeated stimulation. When engineered using targeting moieties of diverse composition, affinity, and specificity, pCAR T cells consistently elicit superior anti-tumor activity compared with T cells that express traditional linear CARs.
KW - cancer
KW - CAR T cells
KW - chimeric antigen receptor
KW - chimeric co-stimulatory receptor
KW - co-stimulation
KW - immunotherapy
KW - parallel CAR
UR - http://www.scopus.com/inward/record.url?scp=85121486222&partnerID=8YFLogxK
U2 - 10.1016/j.xcrm.2021.100457
DO - 10.1016/j.xcrm.2021.100457
M3 - Article
AN - SCOPUS:85121486222
SN - 2666-3791
VL - 2
JO - Cell Reports Medicine
JF - Cell Reports Medicine
IS - 12
M1 - 100457
ER -