Sleeping beauty system to redirect T-cell specificity for human applications

Sourindra N. Maiti, Helen Huls, Harjeet Singh, Margaret Dawson, Matthew Figliola, Simon Olivares, Pullavathi Rao, Yi Jue Zhao, Asha Multani, Ge Yang, Ling Zhang, Denise Crossland, Sonny Ang, Hiroki Torikai, Brian Rabinovich, Dean A. Lee, Partow Kebriaei, Perry Hackett, Richard E. Champlin, Laurence J N Cooper

Research output: Contribution to journalArticlepeer-review

57 Scopus citations

Abstract

The Sleeping Beauty (SB) transposon/transposase DNA plasmid system is used to genetically modify cells for long-term transgene expression. We adapted the SB system for human application and generated T cells expressing a chimeric antigen receptor (CAR) specific for CD19. Electrotransfer of CD19-specific SB DNA plasmids in peripheral blood mononuclear cells and propagation on CD19 artificial antigen presenting cells was used to numerically expand CD3 T cells expressing CAR. By day 28 of coculture, >90% of expanded CD3 T cells expressed CAR. CAR T cells specifically killed CD19 target cells and consisted of subsets expressing biomarkers consistent with central memory, effector memory, and effector phenotypes. CAR T cells contracted numerically in the absence of the CD19 antigen, did not express SB11 transposase, and maintained a polyclonal TCR Vα and TCR Vβ repertoire. Quantitative fluorescence in situ hybridization revealed that CAR T cells preserved the telomere length. Quantitative polymerase chain reaction and fluorescence in situ hybridization showed CAR transposon integrated on average once per T-cell genome. CAR T cells in peripheral blood can be detected by quantitative polymerase chain reaction at a sensitivity of 0.01%. These findings lay the groundwork as the basis of our first-in-human clinical trials of the nonviral SB system for the investigational treatment of CD19 B-cell malignancies (currently under 3 INDs: 14193, 14577, and 14739).

Original languageEnglish (US)
Pages (from-to)112-123
Number of pages12
JournalJournal of Immunotherapy
Volume36
Issue number2
DOIs
StatePublished - Feb 2013

Bibliographical note

Copyright:
Copyright 2013 Elsevier B.V., All rights reserved.

Keywords

  • CD19
  • Sleeping Beauty transposon and transposase
  • T-cell adoptive immunotherapy
  • aAPC
  • chimeric antigen receptor
  • digital mRNA profiling

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