Context-specific synthetic T cell promoters from assembled transcriptional elements

Area of Science:

• Immunology
• Synthetic Biology
• Genetic Engineering

Background:

• Current genetic engineering of human lymphocytes for therapies lacks precise transgene transcriptional control, limiting therapeutic effectiveness.
• The limited understanding of transcriptional logic hinders the rational design of contextually responsive genetic modules for T cell therapies.

Approach:

• Generated a library of inducible synthetic promoters (iSynPros) by random concatemerization of curated transcriptional response elements (TREs).
• Screened the iSynPro library for "IF-THEN" logic-gated transcriptional responses in human CD8+ T cells engineered with a 4-1BB chimeric antigen receptor (CAR).
• Analyzed TRE composition and patterns of selected iSynPros for their ability to regulate antitumor potency enhancement modules.

Key Points:

• Selected iSynPros demonstrated stringent "off-states" in quiescent T cells and CAR activation-dependent transcriptional responsiveness.
• Synthetic TRE grammar was revealed to mediate logic-gated transgene transcription in human T cells.
• The developed iSynPros enhance the potency of CAR T cell engineering.

Conclusions:

• This study presents a novel method for creating context-specific inducible synthetic promoters for T cell engineering.
• The findings demonstrate that synthetic TRE grammar can control logic-gated transgene transcription, enhancing CAR T cell potency and therapeutic indices.