Engineering Human T Cell Receptors for Adoptive T Cell Therapies

Basic Science Poster

A19

Technology

Engineering Human T Cell Receptors for Adoptive T Cell Therapies

Author/Presenter: Daniel Harris
Co-Author(s): None


T cell receptors (TCRs) interact with peptides bound to the major histocompatibility complex (MHC) through the complementary determining regions (CDRs). To better understand the mechanisms governing peptide interactions with TCRs, a switch in specificity of the A6 TCR was engineered from the Tax peptide to the melanoma differentiation antigen MART1 through substitutions of a limited number of CDR residues, and directed evolution. This new TCR, RD1-MART1HIGH, engages MART1/HLA-A2 in an unusual orientation. To identify the CDR residues involved in pepMHC interactions and in peptide specificity, a deep mutational screen was conducted. Single-codon yeast libraries were created for both the A6 and RD1-MART1HIGH TCRs and the libraries were sorted with their respective pepMHC ligands. Deep sequencing analysis was conducted and enrichment/depletion values of mutations were calculated in relation to the pre-sorted population. This analysis showed that the peptide specificity switch could only be achieved by the distributed and collective action of many CDR residues. Substitutions with the highest enrichment values for RD1-MART1HIGH were capable of further improving binding-affinity for MART1/HLA-A2 up to 100-fold. To better understand the signaling capabilities of this novel TCR pepMHC interaction, CD4 and CD8 T cells were transduced with the RD1-MART1HIGH TCR and activity was compared to another MART1 specific TCR with a similar binding KD for MART1/HLA-A2.

Author/Presenter Bio: Dan Harris graduated summa cum laude from Loyola University Chicago in 2011. Upon entering the Medical Scholars Program, he joined Dr. David Kranz’s laboratory. His research focuses on understanding the mechanisms that govern T cell receptor specificity in effort to develop better immunotherapies for the treatment of cancer. When he isn’t studying or working in lab, he enjoys spending time outdoors with his wife Kelly and labrador retriever Murphy.

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