Redirecting T-cell specificity to EGFR using mRNA to self-limit expression of chimeric antigen receptor

Hillary G. Caruso, Hiroki Torikai, Ling Zhang, Sourindra Maiti, Jianliang Dai, Kim Anh Do, Harjeet Singh, Helen Huls, Dean A. Lee, Richard E. Champlin, Amy B. Heimberger, Laurence J.N. Cooper*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

Potential for on-target, but off-tissue toxicity limits therapeutic application of genetically modified T cells constitutively expressing chimeric antigen receptors (CARs) from tumor-associated antigens expressed in normal tissue, such as epidermal growth factor receptor (EGFR). Curtailing expression of CAR through modification of T cells by in vitro-transcribed mRNA species is one strategy to mitigate such toxicity. We evaluated expression of an EGFR-specific CAR coded from introduced mRNA in human T cells numerically expanded ex vivo to clinically significant numbers through coculture with activating and propagating cells (AaPC) derived from K562 preloaded with anti-CD3 antibody. The density of AaPC could be adjusted to affect phenotype of T cells such that reduced ratio of AaPC resulted in higher proportion of CD8+ and central memory T cells that were more conducive to electrotransfer of mRNA than T cells expanded with high ratios of AaPC. RNA-modified CAR+ T cells produced less cytokine, but demonstrated similar cytolytic capacity as DNA-modified CAR+ T cells in response to EGFR-expressing glioblastoma cells. Expression of CAR by mRNA transfer was transient and accelerated by stimulation with cytokine and antigen. Loss of CAR abrogated T-cell function in response to tumor and normal cells expressing EGFR. We describe a clinically applicable method to propagate and modify T cells to transiently express EGFR-specific CAR to target EGFR-expressing tumor cells that may be used to limit on-target, off-tissue toxicity to normal tissue.

Original languageEnglish (US)
Pages (from-to)205-217
Number of pages13
JournalJournal of Immunotherapy
Volume39
Issue number5
DOIs
StatePublished - 2016
Externally publishedYes

Keywords

  • Artificial antigen presenting cells
  • Chimeric antigen receptor
  • EGFR
  • Glioblastoma
  • mRNA

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology
  • Cancer Research

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