A protein-centric approach to understand cancer processes and drugs
Our research aims to improve the understanding of cancer processes by using protein centric strategies. The main method in our lab is the Cellular Thermal Shift Assay (CETSA), the first broadly applicable method to study interactions with proteins in living cells. By using this method we aim to gain knowledge about cellular mechanisms in cancer, discover novel drug targets and find new biomarkers for improved cancer therapy.
Different schemes of CETSA experiments.
To evaluate the protein stability changes using the CETSA principle, the samples used typically differ in the incubated compound (or drug) dose. The 1-dimensional way of CETSA includes classical melting curve scheme of treatment with one compound dose (versus vehicle control) in a range of heating temperatures, and the isothermal dose response (ITDR) scheme of a series of compound doses under one heating temperature (such as the median melting temperature point of the soluble proteome of the studied organism, the typical value for mammalian cell lysate is 52°C and intact-cell is 50°C). The CETSA scheme can be easily expanded to a two-dimensional form with the combination of a range of heating temperatures and compound doses. A most recent addition of 2D-CETSA scheme allows comparison of the protein stability changes and protein level changes between biological samples from different cellular states such as in different cell cycle phases, particularly the profiles of thermal shifts across a range of temperatures are distinctive for many proteins, here we refer to it as IMPRINTS-CETSA.
(Adapted from Dai et al, Annual Rev of Biochem, 2019)
Projects in Stockholm:
Using CETSA to study target engagement of e.g. taxanes in breast cancer.
MS-CETSA as a tool to understand the action of cellular proteomes in basic cancer processes related to tumor immunology.
Proteome-wide mapping using MS-CETSA for several anti-cancer drugs: microtubule inhibitors (e.g. taxanes) and antimetabolite compounds (e.g. 5-Fluorouracil). The project aims at improving our understanding of cancer drug action, discovering novel bio-markers for drug efficacy or toxicity, and identifying mechanisms for innate/acquired drug resistance.
Developing CETSA towards a clinical assay. The project ultimately aims at implementing CETSA as a diagnostic tool to guide personalized cancer therapy. We are currently working with clinical samples from different cancer types (acute myeloid leukemia (AML), breast cancer, and colorectal cancer) for which we are utilizing MS-CETSA to further enhance our knowledge of drug response in the individual patients.
Projects in Singapore:
MS-CETSA for the target de-convolution of BET inhibitors and immunomodulatory drugs
Studying HCV inhibitors with MS-CETSA and the viral infectious process through them
Studying the EMT processes in cancer metastasis
Understanding targets of anti-tubercular drugs using MS-CETSA
Studying resistance mechanisms in cancer cells
Studying drugs affecting ageing processes using IMPRINTS CETSA