piggyBac Transposon/Transposase System to Generate CD19-Specific T Cells for the Treatment of B-Lineage Malignancies

PVR Manuri, MH Wilson, SN Maiti, T Mi… - Human gene …, 2010 - liebertpub.com
PVR Manuri, MH Wilson, SN Maiti, T Mi, H Singh, S Olivares, MJ Dawson, H Huls, DA Lee
Human gene therapy, 2010liebertpub.com
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 …
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.
Mary Ann Liebert