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 journalArticle

51 Citations (Scopus)

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 1 2013

Fingerprint

T-Cell Antigen Receptor Specificity
Beauty
T-Cell Antigen Receptor
Antigen Receptors
CD19 Antigens
Transposases
T-Lymphocytes
Fluorescence In Situ Hybridization
Plasmids
Polymerase Chain Reaction
Investigational Therapies
DNA Transposable Elements
Telomere
Antigen-Presenting Cells
Coculture Techniques
Transgenes
Blood Cells
B-Lymphocytes
Biomarkers
Clinical Trials

Keywords

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

Cite this

Maiti, S. N., Huls, H., Singh, H., Dawson, M., Figliola, M., Olivares, S., ... Cooper, L. J. N. (2013). Sleeping beauty system to redirect T-cell specificity for human applications. Journal of Immunotherapy, 36(2), 112-123. https://doi.org/10.1097/CJI.0b013e3182811ce9

Sleeping beauty system to redirect T-cell specificity for human applications. / Maiti, Sourindra N.; Huls, Helen; Singh, Harjeet; Dawson, Margaret; Figliola, Matthew; Olivares, Simon; Rao, Pullavathi; Zhao, Yi Jue; Multani, Asha; Yang, Ge; Zhang, Ling; Crossland, Denise; Ang, Sonny; Torikai, Hiroki; Rabinovich, Brian; Lee, Dean A.; Kebriaei, Partow; Hackett, Perry; Champlin, Richard E.; Cooper, Laurence J N.

In: Journal of Immunotherapy, Vol. 36, No. 2, 01.02.2013, p. 112-123.

Research output: Contribution to journalArticle

Maiti, SN, Huls, H, Singh, H, Dawson, M, Figliola, M, Olivares, S, Rao, P, Zhao, YJ, Multani, A, Yang, G, Zhang, L, Crossland, D, Ang, S, Torikai, H, Rabinovich, B, Lee, DA, Kebriaei, P, Hackett, P, Champlin, RE & Cooper, LJN 2013, 'Sleeping beauty system to redirect T-cell specificity for human applications', Journal of Immunotherapy, vol. 36, no. 2, pp. 112-123. https://doi.org/10.1097/CJI.0b013e3182811ce9
Maiti SN, Huls H, Singh H, Dawson M, Figliola M, Olivares S et al. Sleeping beauty system to redirect T-cell specificity for human applications. Journal of Immunotherapy. 2013 Feb 1;36(2):112-123. https://doi.org/10.1097/CJI.0b013e3182811ce9
Maiti, Sourindra N. ; Huls, Helen ; Singh, Harjeet ; Dawson, Margaret ; Figliola, Matthew ; Olivares, Simon ; Rao, Pullavathi ; Zhao, Yi Jue ; Multani, Asha ; Yang, Ge ; Zhang, Ling ; Crossland, Denise ; Ang, Sonny ; Torikai, Hiroki ; Rabinovich, Brian ; Lee, Dean A. ; Kebriaei, Partow ; Hackett, Perry ; Champlin, Richard E. ; Cooper, Laurence J N. / Sleeping beauty system to redirect T-cell specificity for human applications. In: Journal of Immunotherapy. 2013 ; Vol. 36, No. 2. pp. 112-123.
@article{b70ae5af82d84cd5904d2acd33e43588,
title = "Sleeping beauty system to redirect T-cell specificity for human applications",
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).",
keywords = "CD19, Sleeping Beauty transposon and transposase, T-cell adoptive immunotherapy, aAPC, chimeric antigen receptor, digital mRNA profiling",
author = "Maiti, {Sourindra N.} and Helen Huls and Harjeet Singh and Margaret Dawson and Matthew Figliola and Simon Olivares and Pullavathi Rao and Zhao, {Yi Jue} and Asha Multani and Ge Yang and Ling Zhang and Denise Crossland and Sonny Ang and Hiroki Torikai and Brian Rabinovich and Lee, {Dean A.} and Partow Kebriaei and Perry Hackett and Champlin, {Richard E.} and Cooper, {Laurence J N}",
year = "2013",
month = "2",
day = "1",
doi = "10.1097/CJI.0b013e3182811ce9",
language = "English (US)",
volume = "36",
pages = "112--123",
journal = "Journal of Immunotherapy",
issn = "1053-8550",
publisher = "Lippincott Williams and Wilkins",
number = "2",

}

TY - JOUR

T1 - Sleeping beauty system to redirect T-cell specificity for human applications

AU - Maiti, Sourindra N.

AU - Huls, Helen

AU - Singh, Harjeet

AU - Dawson, Margaret

AU - Figliola, Matthew

AU - Olivares, Simon

AU - Rao, Pullavathi

AU - Zhao, Yi Jue

AU - Multani, Asha

AU - Yang, Ge

AU - Zhang, Ling

AU - Crossland, Denise

AU - Ang, Sonny

AU - Torikai, Hiroki

AU - Rabinovich, Brian

AU - Lee, Dean A.

AU - Kebriaei, Partow

AU - Hackett, Perry

AU - Champlin, Richard E.

AU - Cooper, Laurence J N

PY - 2013/2/1

Y1 - 2013/2/1

N2 - 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).

AB - 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).

KW - CD19

KW - Sleeping Beauty transposon and transposase

KW - T-cell adoptive immunotherapy

KW - aAPC

KW - chimeric antigen receptor

KW - digital mRNA profiling

UR - http://www.scopus.com/inward/record.url?scp=84873987094&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84873987094&partnerID=8YFLogxK

U2 - 10.1097/CJI.0b013e3182811ce9

DO - 10.1097/CJI.0b013e3182811ce9

M3 - Article

C2 - 23377665

AN - SCOPUS:84873987094

VL - 36

SP - 112

EP - 123

JO - Journal of Immunotherapy

JF - Journal of Immunotherapy

SN - 1053-8550

IS - 2

ER -