Report
Teaser, summary, work performed and final results
Periodic Reporting for period 2 - TagIt (A Minimal-Tag Bioorthogonal Labelling Approach to Protein Uptake, Traffic and Delivery)
Teaser
The ability to probe dynamic cellular events involv disease-associated proteins is limited, to a large degree, by the development of a strategy that uses small-sized coupling partners that react in a selective fashion with very rapid kinetics and without interfering with...
Summary
The ability to probe dynamic cellular events involv disease-associated proteins is limited, to a large degree, by the development of a strategy that uses small-sized coupling partners that react in a selective fashion with very rapid kinetics and without interfering with biological function(s) and localisation. In this ERC StG project, we are developing a conceptually novel bioorthogonal approach by combining the introduction of small alkene-tags with chemoselective reactions that display rapid kinetics to label proteins in live cells. The small size of the handles to be installed on the protein of interest should not interfere with the protein\'s innate function(s) and localisation. Site-selective bioorthogonal labelling will be achieved through the use of a novel reactions that are triggered through the addition of chemical reagents. This approach offers potentially improved spatial and temporal resolution. The proposed methodology will be applied in the context of key actors cancer progression. The ability to label those actors – proteins – with minimal perturbation of its structure, function and localisation will enable time-lapsed monitoring of their uptake into cells and trafficking pathways once inside cells. In doing so, new biological insight into those proteins biology will be generated that will assist the construction of safer, more selective and more efficient protein-drug conjugates for cancer treatment. The concept we are developing has been designed to be generally applicable to label and study disease-associated proteins that are difficult to access using conventional protein labelling methods constituting the first integrated, interdisciplinary approach for the development of protein-drug conjugates. With our proposed strategy we aim to create optimal tissue specific drug-delivery systems that potentiate the efficacy of treatments while minimising side-effects.
Work performed
The main results achieved so far include:
1) the development of a novel bioorthogonal reaction that allow to specifically label a protein of interest in live cells
2) the development of an antibody against the specific receptor (for the protein of interest involved in leukaemia) that enabled to characterise the downstream effects of binding to the the receptor
3) the development of new chemistries that allow the construction of chemically-defined and stable protein conjugates for targeted cancer therapy
4) the award of a proof-of-concept ERC grant to bring discoveries from the bench into the clinic
Final results
During this first stage of the project, we have developed a new chemical tool that allows the chemical installation of a bioorthogonal (i.e., one chemical group that is not present in biological systems) and that react selectively with a non-toxic chemical partner only in the presence of a chemical trigger. The rapid kinetics, chemical-induced nature and biocompatibility of this reaction makes ideal to understand what happens to a protein involved in cancer, for example, once it interacts with receptors on the membrane of the cell.
Until the end of the project, we hope to elucidate the role of one protein involved in leukaemia, and to be able to use it as a delivery agent for potent drugs specifically to cells where the protein receptor is present. This will allow to spare healthy cells from the action of very potent drug, and thus maximise its effect only in cancer cells.
Website & more info
More info: http://www.gbernardeslab.com.