Work performed and main achievements
During the SynT Proof of Concept project, our team achieved a series of key milestones in the development, optimisation, and strategic positioning of a synthetic cis-regulatory DNA in engineered T cell products development. These achievements establish a solid foundation for future translational and collaborative work in oncology and now extend this to autoimmunity.
1. Enhancement of Computational Design Tools
Insights gained during experimental testing informed improvements to our synthetic DNA design platform. A next-generation version of the LSD design suite (LSD+) was assembled, incorporating updated logic for the selection and optimisation of regulatory DNA elements. This software provides enhanced capabilities for future reporter development efforts.
2. Development of Synthetic Reporters for T-Cell Function and Dysfunction
Following the pilot work, using the LSD+ suite, we successfully designed, constructed, and implemented four synthetic locus control region (sLCR)-based reporters reflecting functional activation states and capturing dysfunctional or exhausted states. These reporters constitute the core technological output of SynT and provide the basis for more informative monitoring of engineered immune-cell behaviour.
3. Integration of Reporters into CAR T Cells
Using established CAR architectures, the synthetic reporters were co-transduced into CD4+ T cells together alongside CARs directed against clinically relevant targets. This integration enabled controlled evaluation of reporter activity and laid the groundwork for future applications requiring dynamic readouts of T-cell state.
4. Characterisation Under Defined Experimental Conditions
The reporter cells were analysed under standardised stimulation conditions using flow cytometry and molecular assays. These assays enabled us to test biologically meaningful activation and differentiation cues, a key requisite to assess the feasibility of sLCR-based indicators for monitoring CAR T-cell state transitions.
5. Establishment of New Collaborative Partnerships
To further develop and validate the SynT reporters, formal collaboration agreements were signed with two external institutions specialising in T-cell engineering and validation. These partnerships extend the reach and sustainability of the technology beyond the ERC funding period.
6. Strategic Redirection Toward Autoimmunity Applications
During the project period, major shifts in the CAR T-cell therapy commercial landscape underscored the need for repositioning our long-term strategies. In response, SynT expanded its intended application space toward autoimmune diseases, where selectivity and temporal activation are key. This strategic repositioning broadens the impact and relevance of the SynT technology.
Future Outlook
Building on the SynT proof of concept, future work will focus on refining and extending the reporters within collaborative academic networks and, where appropriate, in structured translational programmes. The broader potential of SynT will continue to be explored through the partnerships and consortium efforts established during the project period, and to align our future commercial strategy with the current drastic shift in the cell therapy industry.
Conclusion
SynT has delivered a comprehensive proof of concept for the use of synthetic regulatory DNA elements to monitor CAR T-cell functional states. The project has strengthened computational tools, generated the intended technological assets, and positioned the technology for continued development across oncology and autoimmunity. These achievements advance the broader goal of creating more precise, predictable, and adaptable engineered immune-cell therapies despite the substantial market pressures.
