piggyBac transposon/transposase system to generate CD19-specific T cells for the treatment of B-lineage malignancies

Hum Gene Ther. 2010 Apr;21(4):427-37. doi: 10.1089/hum.2009.114.

Abstract

Nonviral integrating vectors can be used for expression of therapeutic genes. piggyBac (PB), a transposon/transposase system, has been used to efficiently generate induced pluripotent stems cells from somatic cells, without genetic alteration. In this paper, we apply PB transposition to express a chimeric antigen receptor (CAR) in primary human T cells. We demonstrate that T cells electroporated to introduce the PB transposon and transposase stably express CD19-specific CAR and when cultured on CD19(+) artificial antigen-presenting cells, numerically expand in a CAR-dependent manner, display a phenotype associated with both memory and effector T cell populations, and exhibit CD19-dependent killing of tumor targets. Integration of the PB transposon expressing CAR was not associated with genotoxicity, based on chromosome analysis. PB transposition for generating human T cells with redirected specificity to a desired target such as CD19 is a new genetic approach with therapeutic implications.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antigen-Presenting Cells / immunology
  • Antigens, CD19 / metabolism*
  • Cell Line, Tumor
  • Cells, Cultured
  • Coculture Techniques
  • DNA Transposable Elements* / genetics
  • DNA Transposable Elements* / immunology
  • Electroporation
  • Genetic Therapy / methods
  • Genetic Vectors
  • Glioblastoma
  • Humans
  • K562 Cells
  • Lymphoma, B-Cell / therapy*
  • Plasmids
  • Receptors, Antigen / genetics*
  • Receptors, Antigen / metabolism
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism
  • Transgenes
  • Transposases* / genetics
  • Transposases* / immunology

Substances

  • Antigens, CD19
  • DNA Transposable Elements
  • Receptors, Antigen
  • Transposases