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Imaging of Sleeping Beauty-Modified CD19-Specific T Cells Expressing HSV1-Thymidine Kinase by Positron Emission Tomography

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Abstract

Purpose

We have incorporated a positron emission tomography (PET) functionality in T cells expressing a CD19-specific chimeric antigen receptor (CAR) to non-invasively monitor the adoptively transferred cells.

Procedures

We engineered T cells to express CD19-specific CAR, firefly luciferase (ffLuc), and herpes simplex virus type-1 thymidine kinase (TK) using the non-viral-based Sleeping Beauty (SB) transposon/transposase system adapted for human application. Electroporated primary T cells were propagated on CD19+ artificial antigen-presenting cells.

Results

After 4 weeks, 90 % of cultured cells exhibited specific killing of CD19+ targets in vitro, could be ablated by ganciclovir, and were detected in vivo by bioluminescent imaging and PET following injection of 2′-deoxy-2′-[18F]fluoro-5-ethyl-1-β-d-arabinofuranosyl-uracil ([18F]FEAU).

Conclusion

This is the first report demonstrating the use of SB transposition to generate T cells which may be detected using PET laying the foundation for imaging the distribution and trafficking of T cells in patients treated for B cell malignancies.

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Abbreviations

[18F]-FEAU:

2′-deoxy-2′-[18F]fluoro-5-ethyl-1-β-d-arabinofuranosyl-uracil

[18F]-FHBG:

9-(4-[18F]fluoro-3-hydroxymethylbutyl)guanine

aAPC:

Artificial presenting cells, K562-derived

BLI:

Bioluminescent imaging

CAR:

Chimeric antigen receptor

CoOp:

Codon optimized

CRA:

Chromium release assay

CT:

Computed tomography

GCV:

Ganciclovir

HSCT:

Hematopoietic stem-cell transplantation

HSV-1:

Herpes simplex virus-1

Hygro:

Hygromycin phosphotransferase

mIL-15:

Membrane-bound interleukin-15

Neo:

Neomycin phosphotransferase

NES:

Nuclear export signal

NLS:

Nuclear localization sequence

NLSm:

Nuclear localization sequence, mutated

PET:

Positron emission tomography

PBMC:

Peripheral blood mononuclear cells

SB:

Sleeping Beauty

TAA:

Tumor-associated antigen

[14C]-Thd:

[14C]thymidine

TK:

Thymidine kinase, herpes simplex virus type-1

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Acknowledgments

The authors would like to thank Dr. Carl June (University of Pennsylvania) for help in generating and providing aAPC clone no. 4 and Dr. Perry Hackett (University of Minnesota) for help with the SB system. Grant support includes R01 (CA124782, CA120956, CA141303), R33 (CA116127), P01 (CA148600), Albert J Ward Foundation, Burroughs Wellcome Fund, Gillson Longenbaugh Foundation, Cancer Prevention and Research Institute of Texas, CLL Global Research Foundation, National Foundation for Cancer Research, and Pediatric Cancer Research Foundation.

Author information

Author notes

  1. Dean A. Lee

    Present address: The University of Texas Graduate School of Biomedical Sciences, Houston, TX, USA

  2. Laurence J.N. Cooper

    Present address: Ziopharm Oncology Inc., Boston, MA, USA

Authors and Affiliations

  1. Division of Pediatrics, The University of Texas M.D. Anderson Cancer Center, Houston, TX, USA

    Amer M. Najjar, Simon Olivares, Tiejuan Mi, Helen Huls, Brian Rabinovich, Dean A. Lee & Laurence J.N. Cooper

  2. Melanoma Medical Oncology-Research, The University of Texas M.D. Anderson Cancer Center, Houston, TX, USA

    Pallavi R Manuri & Jason Roszik

  3. Department of Cancer Systems Imaging, The University of Texas M.D. Anderson Cancer Center, Houston, TX, USA

    Leo Flores II & Mian Alauddin

  4. Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas M.D. Anderson Cancer Center, Houston, TX, USA

    Elizabeth J. Shpall & Richard E. Champlin

  5. Molecular Imaging Program, Karmanos Cancer Institute, Wayne State University, Detroit, MI, USA

    Nashaat Turkman & Juri Gelovani

  6. Center for Advanced Biomedical Imaging, The University of Texas M.D. Anderson Cancer Center, Houston, TX, USA

    Vincenzo Paolillo

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  1. Amer M. Najjar
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Corresponding author

Correspondence to Laurence J.N. Cooper.

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Conflict of Interest

The technology described in this publication was advanced through research conducted at the University of Texas MD Anderson Cancer Center (MD Anderson) by Laurence Cooper. In January 2015, the technology was licensed by MD Anderson for commercial application to ZIOPHARM Oncology, Inc., and Intrexon Corporation, in exchange for equity interests in each of these companies. Laurence Cooper, Amer M. Najjar, Pallavi R. Manuri, Simon Olivares, Tiejuan Mi, Helen Huls, Richard E. Champlin, Brian Rabinovich, and Dean A. Lee are eligible in accordance with the Rules of Board of Regents of The University of Texas System to share in the proceeds of the disposition of the equity received by MD Anderson as a result of the licensing of this technology. On May 7, 2015, Dr. Cooper was appointed as the Chief Executive Officer at ZIOPHARM. Dr. Cooper is now a Visiting Scientist at MD Anderson where he continues to help supervise the development of this technology. The research being reported in this publication is research in which The University of Texas MD Anderson Cancer Center has an institutional financial conflict of interest. Because MD Anderson is committed to the protection of human subjects and the effective management of its financial conflicts of interest in relation to its research activities, MD Anderson has implemented an Institutional Conflict of Interest Management and Monitoring Plan to manage and monitor the conflict of interest with respect to MD Anderson’ s conduct of this research.

Additional information

Amer M. Najjar, Pallavi R Manuri and Simon Olivares contributed equally to this work.

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Najjar, A.M., Manuri, P.R., Olivares, S. et al. Imaging of Sleeping Beauty-Modified CD19-Specific T Cells Expressing HSV1-Thymidine Kinase by Positron Emission Tomography. Mol Imaging Biol 18, 838–848 (2016). https://doi.org/10.1007/s11307-016-0971-8

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  • DOI: https://doi.org/10.1007/s11307-016-0971-8

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