Biomedical Microdevices

, Volume 12, Issue 5, pp 855–863

A high throughput microelectroporation device to introduce a chimeric antigen receptor to redirect the specificity of human T cells

  • Yoonsu Choi
  • Carrie Yuen
  • Sourindra N. Maiti
  • Simon Olivares
  • Hillary Gibbons
  • Helen Huls
  • Robert Raphael
  • Thomas C. Killian
  • Daniel J. Stark
  • Dean A. Lee
  • Hiroki Torikai
  • Daniel Monticello
  • Susan S. Kelly
  • Partow Kebriaei
  • Richard E. Champlin
  • Sibani L. Biswal
  • Laurence J. N. Cooper
Article

DOI: 10.1007/s10544-010-9440-3

Cite this article as:
Choi, Y., Yuen, C., Maiti, S.N. et al. Biomed Microdevices (2010) 12: 855. doi:10.1007/s10544-010-9440-3

Abstract

It has been demonstrated that a chimeric antigen receptor (CAR) can directly recognize the CD19 molecule expressed on the cell surface of B-cell malignancies independent of major histocompatibility complex (MHC). Although T-cell therapy of tumors using CD19-specific CAR is promising, this approach relies on using expression vectors that stably integrate the CAR into T-cell chromosomes. To circumvent the potential genotoxicity that may occur from expressing integrating transgenes, we have expressed the CD19-specific CAR transgene from mRNA using a high throughput microelectroporation device. This research was accomplished using a microelectroporator to achieve efficient and high throughput non-viral gene transfer of in vitro transcribed CAR mRNA into human T cells that had been numerically expanded ex vivo. Electro-transfer of mRNA avoids the potential genotoxicity associated with vector and transgene integration and the high throughput capacity overcomes the expected transient CAR expression, as repeated rounds of electroporation can replace T cells that have lost transgene expression. We fabricated and tested a high throughput microelectroporator that can electroporate a stream of 2 × 108 primary T cells within 10 min. After electroporation, up to 80% of the passaged T cells expressed the CD19-specific CAR. Video time-lapse microscopy (VTLM) demonstrated the redirected effector function of the genetically manipulated T cells to specifically lyse CD19+ tumor cells. Our biomedical microdevice, in which T cells are transiently and safely modified to be tumor-specific and then can be re-infused, offers a method for redirecting T-cell specificity, that has implications for the development of adoptive immunotherapy.

Keywords

ElectroporationCancerHigh throughputmRNAChimeric antigen receptorT cells

Supplementary material

View video
Supplementary material 1

Video of the electro-transfer process of CAR transgene mRNA using HiTMeD. Extended cables deliver the electrical signal to HiTMeD that operates in a fume hood to maintain sterility. Sampling of electroporated cells is performed using multi-well plates. (MPG 4323 kb)

View video
Supplementary material 2

Time lapse video microscopy redirected killing of adherent tumor targets by CAR+ T cells after electro-transfer. (MPG 4035 kb)

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Yoonsu Choi
    • 1
  • Carrie Yuen
    • 1
  • Sourindra N. Maiti
    • 1
  • Simon Olivares
    • 1
  • Hillary Gibbons
    • 1
  • Helen Huls
    • 1
  • Robert Raphael
    • 4
  • Thomas C. Killian
    • 5
  • Daniel J. Stark
    • 5
  • Dean A. Lee
    • 1
  • Hiroki Torikai
    • 1
  • Daniel Monticello
    • 6
  • Susan S. Kelly
    • 1
  • Partow Kebriaei
    • 2
  • Richard E. Champlin
    • 2
  • Sibani L. Biswal
    • 3
  • Laurence J. N. Cooper
    • 1
  1. 1.Division of Pediatrics, Children’s Cancer Hospital, The University of Texas Graduate School of Biomedical Sciences at HoustonThe University of Texas M. D. Anderson Cancer CenterHoustonUSA
  2. 2.Department of Stem Cell Transplantation and Cellular TherapyThe University of Texas M. D. Anderson Cancer CenterHoustonUSA
  3. 3.Department of Chemical and Biomolecular EngineeringRice UniversityHoustonUSA
  4. 4.Department of BioengineeringRice UniversityHoustonUSA
  5. 5.Department of Physics and AstronomyRice UniversityHoustonUSA
  6. 6.InCellerate Inc.HoustonUSA