Periodic Reporting for period 4 - TagIt (A Minimal-Tag Bioorthogonal Labelling Approach to Protein Uptake, Traffic and Delivery)
Reporting period: 2021-04-01 to 2023-03-31
Our research and approach is important because it allows an informed design of conjugates that are able to deliver a toxic payload to cancer cells while minimising adverse side-effects.
In the context of our research, we also developed a new self-immolative linker that is specifically cleaved in cancer cells. This new linker when combined with the specificity of antibodies (or other ligands) for cancer cells allows the construction of highly selective conjugates for cancer therapy.
Overall, our approach project highlighted how chemistry-driven approaches improve our understanding of complex biological phenomena that will inform the next generation of therapeutics.
1) the development of a novel bioorthogonal reaction that allow to specifically label a protein of interest in live cells. In particular, we developed a new biorthogonal labelling reaction between small-sized alkyne trifluoroborates and tetrazines that has allowed us to characterise the uptake and intracellular trafficking of a target receptor in leukemias;
2) the development of an antibody against the leukemia-specific receptor that enabled to characterise the downstream effects of binding to the the receptor;
3) we developed a new-self immolative linker that allows the cleavable of a C–C bond to release intracellularly an ortho-quinone drug;
4) we built conjugates that were shown to selectively induce cancer cell death both in cells and mouse models of cancer;
5) this research lead to discoveries that are the basis of the award of two proof-of-concept ERC grants to bring discoveries from the bench into the clinic.
Exploitation and dissemination of the results of the project included peer-reviewed publications in high impact journals, presentations at conferences and IP protection (which is now licensed). In addition, I have participated in a number of events in secondary schools where I discuss the role of basic science in health and the need for interdisciplinary approaches to major research questions.
1) first, we developed the first on-demand activable bioorthogonal reaction. This means, we can trigger the reactivity to allow a super fast reaction (complete within 9 seconds) that does not interfere with endogenous cellular processes
2) secondly, and in our search for the best linker+payload, we ended up developing a new self-immolative linker strategy for the protection of ortho-quinone payloads. Importantly, we showed for the first time that a C–C bond may be cleaved in vivo through a 1,6-elimination reaction.
Chemistry is at the center of our research. While the bioorthogonal chemistry provided insight into the internalisation and intracellular trafficking of a relevant receptor overexpressed in cancer cells, the second allows for specific intracellular releaase of a drug once the complex antibody/receptor is internalised. This combination led to the construction of conjugates which we showed to be safe and highly efficacious in relevant models of leukemia.