Description
Transgenic expression of antigen-specific T cell receptor (TCR) genes is a promising approach for immunotherapy against infectious diseases and cancers. A key to the efficient application of this approach is the rapid and specific isolation and cloning of TCRs. Researchers at St. Jude have developed a novel method to rapidly clone, express and characterize the function of paired αβ and γδ TCR chains from single cells. The platform addresses the non-specific, labor-intensive, and time-consuming issues of traditional PCR-based cloning and it provides a relatively high-throughput, accurate, and efficient method of TCR engineering for therapeutic or research applications.
The researchers demonstrated the capability of cloning influenza-specific TCRs within 10 days using single cell PCR and Gibson Assembly techniques. This process can be accelerated to 5 days by generating receptor libraries, requiring only the exchange of the antigen-specific CDR3 region into an existing backbone. The functional activity of these TCRs can be characterized in a novel reporter cell line for screening of TCR specificity and avidity.
By generating a library of specific TCR constructs reactive against a range of viruses and HLA types, TCR-directed therapies could be used prophylactically or immediately at the earliest signs of viral reactivation or to target conserved or patient-specific tumor antigens.
In addition to therapeutic applications, the protocol significantly improves the workflow for cloning and expressing TCRs for study in vitro.
Keywords
Paired T-cell receptors (TCRs), TCR screening, stem cell transplant
Granted Patents or Published Applications
U. S. Patent application submitted, published international application WO 2017/096239
Related Scientific References
Xi-zhi J Guo, Pradyot Dash, Matthew Calverley, Suzanne Tomchuck, Mari H Dallas, and Paul G Thomas, Rapid cloning, expression, and functional characterization of paired αβ and γδ T-cell receptor chains from single-cell analysis. Mol Ther Methods Clin Dev. 2016; 3: 15054. Published online 2016 Jan 27.
Wang, G.C., Dash, P., McCullers, J.A., Doherty, P.C., and Thomas, P.G., T cell receptor αβ diversity inversely correlates with pathogen-specific antibody levels in human cytomegalovirus infection. Sci Transl Med 4, 128ra42 (2012).
Han, A et al., Linking T-cell receptor sequence to functional phenotype at the single cell level. Nature Biotech. 32(7): 684-692 (epub June 22, 2014).
Licensing Opportunities
This invention enables a reduction in cost, time and reagents can get the cost per cell to under $1 per cell, compared with the standard sequencing technique which is now typically $6 per cell, it may be able to be used for cytomegalovirus (CMV) or other herpesvirus-positive patients and cancer patients in patient specific therapy, or it may be used as a molecular tool (i.e. metrics of immune health; better indicator of potent immune status) for drug development. If you are interested in licensing this technology in one of these or any other fields, please contact chad.riggs@stjude.org.
Related Links
- Activation of Prodrugs by Carboxylesterase (SJ-98-0001)
- Anti-GD2-BB-zeta Chimeric Receptor for Treating GD2+ Malignancies (SJ-13-0035)
- Antibodies to Tim4 for use as an Immune Enhancer and Cancer Therapeutic (SJ-18-0006)
- Association of Neuraminidase Regulated Exocytosis with Cancer Metastasis (SJ-11-0012)
- BCRP/ABCG2 as Stem Cell Marker (SJ-97-0016)
- CD33 CAR (SJ-17-0010)
- CD7 CAR placed in CD7neg memory cells (SJ-15-0020)
- Chimeric gene and protein that can be used to create Optogranules: Light induced stress granules (SJ-18-0010)
- Development and Optimization of a Serotype-Independent Method of Adeno-Associated Virus (AAV) Harvest and Purification (SJ-16-0036)
- DNA Methylation Profile and Biomarkers for identifying functional T-cells (SJ-17-0002)
- DNMT3a knockout CAR T cells with Antigen Specificity (for solid tumors) (SJ-19-0024)
- Efficient generation of T-Cell Receptor (TCR) sequences in response to a variety of immune responses (SJ-19-0017)
- Erythroid Specific Promoter for Hematopoietic Disorders (SJ-16-0040)
- Gene Therapy for Wiskott-Aldrich Syndrome (SJ-19-0012)
- Generation of Therapeutic T Cell Receptors for Fibrolamellar Cancer (SJ-19-0046)
- HMGA2 in Gene Therapy Vectors (SJ-16-0014)
- Hybrid Compounds to treat Gastrointestinal Infections (SJ-14-0019/UTA 14-01)
- IL35 Receptor (SJ-08-0039)
- Immune Cells with DNMT3A Gene Modifications (SJ-16-0009)
- Improved GD2 Ab for Neuroblastoma (SJ-17-0017)
- Improved Method to Produce Proteins to Treat Lysosomal Storage Disorders (SJ-01-0020)
- Interleukin-35 (IL-35) (SJ-06-0016)
- Method for Enhancing Recombinant Antibody Production (SJ-08-0032)
- Method for predicting T-cell development stage (and therapy success) (SJ-19-0033)
- Minimized Liver Specific Promoter for Improved Gene Expression in Gene Therapy Vectors (SJ-04-0024)
- miRNAs for Treating Cerebral Ventricle Enlargement in Schizophrenia Patients (SJ-19-0039)
- Myd88 and CD40 Costimulatory Domains for Chimeric Antigen Receptors (SJ-18-0021)
- New Method for Differentiating T-cells (SJ-18-0019)
- p53 Inhibitory Oligonucloeotides (SJ-09-0015)
- Protease Serine 21 (PRSS21) Targeted Immunotherapies (SJ-18-0043)
- REGNASE-1 Gene Knockout or BATF Overexpression for Improving T-cell Function (SJ-19-0027, SJ-20-0007)
- Regulating p53 Translation and Function (SJ-05-0002)
- RIPK3 Fusion Protein for Treating Cancer and Autoimmunity (SJ-12-0036)
- Safety Test for Gene Therapy Vectors (SJ-15-0027)
Contact the Office of Technology Licensing (Phone: 901-595-2342, Fax: 901-595-3148) for more information.