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

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.

• Authors: Pallavi V Raja Manuri, Matthew H Wilson, Sourindra N Maiti, Tiejuan Mi, Harjeet Singh, Simon Olivares, Margaret J Dawson, Helen Huls, Dean A Lee, Pulivarthi H Rao, Joseph M Kaminski, Yozo Nakazawa, Stephen Gottschalk, Partow Kebriaei, Elizabeth J Shpall, Richard E Champlin, Laurence J N Cooper
• Journal: Human gene therapy (Hum Gene Ther) Vol. 21 Issue 4 Pg. 427-37 (Apr 2010) ISSN: 1557-7422 [Electronic] United States
• PMID: 19905893 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

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

Topics

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