Steady and promising advances have been made over the last two decades in chimeric antigen receptor (CAR)-mediated T cell (T-body) therapy. These efforts have led to major breakthroughs in the treatment of B cell malignancies and neuroblastomas in recent clinical trials. However, broader and more rapid implementation of this promising immunotherapy modality has been hindered by the labor-intensive, time-consuming, and expensive ex vivo T cell modification and expansion procedures. Furthermore, monitoring of therapeutic efficacy during the course of treatment remains suboptimal, relying on peripheral blood sampling methods that provide only a snapshot of therapeutic efficacy. To address these logistical and therapy management limitations, we are proposing to develop a combined gene delivery/imaging system based on targeting lentivectors that can specifically transduce T cells in situ with tumor-antigen specific CAR genes and herpes simplex virus-1 thymidine kinase (HSV1tk), a positron emission tomography reporter (PET) gene. This direct gene delivery approach will eliminate the need to genetically modify and expand T cells ex vivo and will provide a means of noninvasively monitoring the biodistribution, trafficking, and targeting of T-bodies to tumor tissue by PET using the nucleoside analog substrate 2'-deoxy-2'-[18F]fluoro-5-ethyl-1-?-D-arabinofuranosyl- uracil ([18F]-FEAU). Expression of HSV1tk will also enable selective ablation of transduced cells in the event of any adverse immune response. Our unique multifunctional gene delivery/imaging system is based on the central hypothesis that direct and specific in situ delivery of tumor antigen-specific CAR and HSV1tk to endogenous T cells by lentiviral vectors (via CD3-binding receptors) will redirect their specificity toward tumo cells and enable noninvasive and repetitive imaging of their targeting patterns by PET. This subset of T-bodies will further expand upon contact with target antigen. Our long-term goal is to develop an "off-the-shelf" lentiviral system that can be customized to express any tumor antigen-specific CAR and a PET reporter/suicide gene to inject into patients to redirect their T cells toward tumor cells while monitoring therapeutic progress by PET. As a step toward achieving our long-term goal, we are pursuing this multi-PI R01 funding mechanism to develop a prototype lentivector that will deliver erbB-2-specific CAR and HSV1tk to endogenous T cells in a mouse breast cancer model. A diverse team of investigators, headed by Professor Zelig Eshhar (PI) who pioneered CAR-based immunotherapy, will contribute expertise in the fields of CAR development, T cell targeting, and PET imaging to develop this multifunctional gene delivery/imaging therapeutic modality. Successful completion of the proposed work will have a significant impact on the implementation of cancer immunotherapy providing a larger number of patients access to promising CAR-based immunotherapy trials and expanding the application of immunotherapy to a host of malignancies.

Public Health Relevance

Immunotherapy of cancer using reprogrammed T cells that target tumors is a promising new approach that is increasingly being applied. Current methods of preparing and growing these tumor-targeting T cells are labor- intensive, time-consuming, and expensive, limiting the number of patients who can receive this potentially curative therapy. We are proposing the development of a direct T cell reprogramming procedure that utilizes a lentivirus vector to directly reprogram patient T cells to target tumor cells without the need to perform any external T cell manipulations. This direct form of T cell preparation will provide a larger number of patients access to potentially curative immunotherapy trials.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB017206-02
Application #
8725662
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Tucker, Jessica
Project Start
2013-09-01
Project End
2017-08-31
Budget Start
2014-09-01
Budget End
2015-08-31
Support Year
2
Fiscal Year
2014
Total Cost
$426,075
Indirect Cost
$31,561
Name
Tel-Aviv Sourasky Medical Center
Department
Type
DUNS #
600121230
City
Tel Aviv
State
Country
Israel
Zip Code
64239
Liu, Yarong; Joo, Kye-Il; Lei, Yuning et al. (2014) Visualization of intracellular pathways of engineered baculovirus in mammalian cells. Virus Res 181:81-91