Redirecting specificity of T-cell populations for CD19 using the sleeping beauty system

Harjeet Singh, Pallavi R. Manuri, Simon Olivares, Navid Dara, Margaret J. Dawson, Helen Huls, Perry B. Hackett, Donald B. Kohn, Elizabeth J. Shpall, Richard E. Champlin, Laurence J.N. Cooper

Research output: Contribution to journalArticle

180 Citations (Scopus)

Abstract

Genetic modification of clinical-grade T cells is undertaken to augment function, including redirecting specificity for desired antigen. We and others have introduced a chimeric antigen receptor (CAR) to enable T cells to recognize lineage-specific tumor antigen, such as CD19, and early-phase human trials are currently assessing safety and feasibility. However, a significant barrier to next-generation clinical studies is developing a suitable CAR expression vector capable of genetically modifying a broad population of T cells. Transduction of T cells is relatively efficient but it requires specialized manufacture of expensive clinical grade recombinant virus. Electrotransfer of naked DNA plasmid offers a cost-effective alternative approach, but the inefficiency of transgene integration mandates ex vivo selection under cytocidal concentrations of drug to enforce expression of selection genes to achieve clinically meaningful numbers of CAR+ T cells. We report a new approach to efficiently generating T cells with redirected specificity, introducing DNA plasmids from the Sleeping Beauty transposon/transposase system to directly express a CD19-specific CAR in memory and effector T cells without drug selection. When coupled with numerical expansion on CD19+ artificial antigen-presenting cells, this gene transfer method results in rapid outgrowth of CD4+ and CD8+ T cells expressing CAR to redirect specificity for CD19+ tumor cells.

Original languageEnglish (US)
Pages (from-to)2961-2971
Number of pages11
JournalCancer Research
Volume68
Issue number8
DOIs
StatePublished - Apr 15 2008

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T-Cell Antigen Receptor Specificity
Beauty
T-Lymphocytes
Antigen Receptors
Population
CD19 Antigens
Plasmids
Transposases
DNA
Neoplasm Antigens
Antigen-Presenting Cells
T-Cell Antigen Receptor
Transgenes
Pharmaceutical Preparations
Viruses
Safety
Gene Expression
Antigens
Costs and Cost Analysis

Cite this

Singh, H., Manuri, P. R., Olivares, S., Dara, N., Dawson, M. J., Huls, H., ... Cooper, L. J. N. (2008). Redirecting specificity of T-cell populations for CD19 using the sleeping beauty system. Cancer Research, 68(8), 2961-2971. https://doi.org/10.1158/0008-5472.CAN-07-5600

Redirecting specificity of T-cell populations for CD19 using the sleeping beauty system. / Singh, Harjeet; Manuri, Pallavi R.; Olivares, Simon; Dara, Navid; Dawson, Margaret J.; Huls, Helen; Hackett, Perry B.; Kohn, Donald B.; Shpall, Elizabeth J.; Champlin, Richard E.; Cooper, Laurence J.N.

In: Cancer Research, Vol. 68, No. 8, 15.04.2008, p. 2961-2971.

Research output: Contribution to journalArticle

Singh, H, Manuri, PR, Olivares, S, Dara, N, Dawson, MJ, Huls, H, Hackett, PB, Kohn, DB, Shpall, EJ, Champlin, RE & Cooper, LJN 2008, 'Redirecting specificity of T-cell populations for CD19 using the sleeping beauty system', Cancer Research, vol. 68, no. 8, pp. 2961-2971. https://doi.org/10.1158/0008-5472.CAN-07-5600
Singh H, Manuri PR, Olivares S, Dara N, Dawson MJ, Huls H et al. Redirecting specificity of T-cell populations for CD19 using the sleeping beauty system. Cancer Research. 2008 Apr 15;68(8):2961-2971. https://doi.org/10.1158/0008-5472.CAN-07-5600
Singh, Harjeet ; Manuri, Pallavi R. ; Olivares, Simon ; Dara, Navid ; Dawson, Margaret J. ; Huls, Helen ; Hackett, Perry B. ; Kohn, Donald B. ; Shpall, Elizabeth J. ; Champlin, Richard E. ; Cooper, Laurence J.N. / Redirecting specificity of T-cell populations for CD19 using the sleeping beauty system. In: Cancer Research. 2008 ; Vol. 68, No. 8. pp. 2961-2971.
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