Pär Nordlund's Group
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 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.
Proteins are targets for the majority of drugs and are involved in most cellular process. However, to study proteins and their interactions in live cells and tissues has been very challenging. Recently our group introduced a new method to meet this challenge: the Cellular Thermal Shift Assay (CETSA) (Science 341(6141):84-87). CETSA is a biophysical assay based on the principle of ligand-induced thermal stabilization of target proteins, meaning that a proteins melting temperature will change upon ligand interaction. Thus, by heating samples (lysate, cells or tissue pieces) to different temperatures, and quantifying proteins in the soluble fraction we can detect altered protein interactions after for example drug treatment. This can either be done for selected proteins of interest by using anti-body based assays or proteome wide by using mass spec. CETSA allows direct monitoring of ligand binding to a specific target (target engagement) in lysate, live cells or even tissue pieces. It can also be used to study downstream effects on protein interaction, providing a novel perspective on protein function in situ.
Using CETSA to study cancer
The current focus in our lab is to explore how CETSA can be used in order to improve cancer treatment. We study key processes in cancer cells as well as the effect of drug treatment on protein interactions. Today, a main drawback of many cytostatics is innate or acquired drug resistance. Thus, we also work on gaining knowledge about the cellular mechanisms during resistance development. These approaches will hopefully lead to the discovery of new biomarkers that in the future can be used to predict treatment outcome and help guiding personalized medicine.
Some of our projects also include studies on tumour associated immune cells and the interaction between the immune system and tumour cells. Novel findings in the immune-oncology field have raised hope that many previously untreatable cancers might be possible to target. For some patient groups such treatment may represent actual cures. However, a large proportion of patients does not respond to treatment or develop resistance. Thus, there is a large interest to gain a better understanding of the intricate interplay between the immune system and the tumour cells. Improved immunotherapies could be a game changer in cancer research and we believe that CETSA has the potential to contribute to this goal.
Taken together, we use CETSA to:
- Provide detailed mechanistic understanding of cellular proteomics in basic cancer processes
- Gain a better understanding of protein interactions in cancer-related immune processes
- Provide in-depth mechanistic understanding of resistance to cytostatics and immunotherapies
- Identify novel candidate drug targets
- Find new protein-based biomarkers that report on therapeutic efficacy and resistance that can help guide personalized therapy
In summary, we use CETSA to study protein interactions in order to acquire data that in has the potential to improve future cancer treatment.
Pär Nordlund, Professor
Smaranda Bacanu, PhD student
Mindaugas Raitelaitis, PhD student
Anette Langebäck, Laboratory coordinator
Henritte Laursen, Post doc
Sara Lööf, Project manager
Anderson Ramos, Post doc
Lu Wang, Post doc
- Knut och Alice Wallenbergs stiftelse
- Vetenskapsrådet Medicin och Hälsa
Horizontal Cell Biology: Monitoring Global Changes of Protein Interaction States with the Proteome-Wide Cellular Thermal Shift Assay (CETSA).
Dai L, Prabhu N, Yu LY, Bacanu S, Ramos AD, Nordlund P
Annu. Rev. Biochem. 2019 Jun;88():383-408
Modulation of Protein-Interaction States through the Cell Cycle.
Dai L, Zhao T, Bisteau X, Sun W, Prabhu N, Lim Y, et al
Cell 2018 05;173(6):1481-1494.e13
Thermal proximity coaggregation for system-wide profiling of protein complex dynamics in cells.
Tan CSH, Go KD, Bisteau X, Dai L, Yong CH, Prabhu N, et al
Science 2018 03;359(6380):1170-1177
Dual blockade of the lipid kinase PIP4Ks and mitotic pathways leads to cancer-selective lethality.
Kitagawa M, Liao PJ, Lee KH, Wong J, Shang SC, Minami N, et al
Nat Commun 2017 12;8(1):2200
CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil.
Almqvist H, Axelsson H, Jafari R, Dan C, Mateus A, Haraldsson M, et al
Nat Commun 2016 Mar;7():11040
The Cellular Thermal Shift Assay: A Novel Biophysical Assay for In Situ Drug Target Engagement and Mechanistic Biomarker Studies.
Martinez Molina D, Nordlund P
Annu. Rev. Pharmacol. Toxicol. 2016 ;56():141-61
The cellular thermal shift assay for evaluating drug target interactions in cells.
Jafari R, Almqvist H, Axelsson H, Ignatushchenko M, Lundbäck T, Nordlund P, et al
Nat Protoc 2014 Sep;9(9):2100-22
Tracking cancer drugs in living cells by thermal profiling of the proteome.
Savitski MM, Reinhard FB, Franken H, Werner T, Savitski MF, Eberhard D, et al
Science 2014 Oct;346(6205):1255784
Engineering protein thermostability using a generic activity-independent biophysical screen inside the cell.
Asial I, Cheng YX, Engman H, Dollhopf M, Wu B, Nordlund P, et al
Nat Commun 2013 ;4():2901
Monitoring drug target engagement in cells and tissues using the cellular thermal shift assay.
Martinez Molina D, Jafari R, Ignatushchenko M, Seki T, Larsson EA, Dan C, et al
Science 2013 Jul;341(6141):84-7