2. Academic Validation
  3. 4-1BB-encoding CAR causes cell death via sequestration of the ubiquitin-modifying enzyme A20

4-1BB-encoding CAR causes cell death via sequestration of the ubiquitin-modifying enzyme A20

  • Cell Mol Immunol. 2024 Jun 27. doi: 10.1038/s41423-024-01198-y.
Zhangqi Dou 1 2,  Thomas Raphael Bonacci 3,  Peishun Shou 1,  Elisa Landoni 1,  Mark G Woodcock 1 4,  Chuang Sun 1,  Barbara Savoldo 1 5,  Laura E Herring 6,  Michael J Emanuele 3,  Feifei Song 1,  Albert S Baldwin 1 3,  Yisong Wan 1 7,  Gianpietro Dotti 8 9,  Xin Zhou 10 11
Affiliations

Affiliations

  • 1 Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, USA.
  • 2 Department of Neurosurgery, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.
  • 3 Department of Genetics, University of North Carolina, Chapel Hill, NC, USA.
  • 4 Division of Oncology, Department of Medicine, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
  • 5 Department of Pediatrics, University of North Carolina, Chapel Hill, NC, USA.
  • 6 Michael Hooker Proteomics Center, Department of Pharmacology, University of North Carolina, Chapel Hill, NC, USA.
  • 7 Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, NC, USA.
  • 8 Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, USA. gdotti@med.unc.edu.
  • 9 Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, NC, USA. gdotti@med.unc.edu.
  • 10 Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, USA. zhouxin@email.unc.edu.
  • 11 Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, NC, USA. zhouxin@email.unc.edu.
PMID: 38937625 DOI: 10.1038/s41423-024-01198-y
Abstract

CD28 and 4-1BB costimulatory endodomains included in chimeric antigen receptor (CAR) molecules play a critical role in promoting sustained antitumor activity of CAR-T cells. However, the molecular events associated with the ectopic and constitutive display of either CD28 or 4-1BB in CAR-T cells have been only partially explored. In the current study, we demonstrated that 4-1BB incorporated within the CAR leads to cell cluster formation and cell death in the forms of both Apoptosis and Necroptosis in the absence of CAR tonic signaling. Mechanistic studies illustrate that 4-1BB sequesters A20 to the cell membrane in a TRAF-dependent manner causing A20 functional deficiency that in turn leads to NF-κB hyperactivity, cell aggregation via ICAM-1 overexpression, and cell death including Necroptosis via RIPK1/RIPK3/MLKL pathway. Genetic modulations obtained by either overexpressing A20 or releasing A20 from 4-1BB by deleting the TRAF-binding motifs of 4-1BB rescue cell cluster formation and cell death and enhance the antitumor ability of 4-1BB-costimulated CAR-T cells.

Keywords

4-1BB; A20; Chimeric antigen receptor (CAR)-T cell; NF-κB; Necroptosis; TNF receptor associated factor (TRAF).

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