Probing DNA-damage response at the single cell level

Prof. Thalassinos's research focuses on understanding how cells respond to DNA damage using single-cell proteomics. The study employs a compound called 60G to induce DNA damage, which activates DNA repair mechanisms such as mismatch repair, base excision repair, and nucleotide excision repair. By analyzing single cells, the research aims to uncover the variability in protein expression and the biological processes involved in DNA repair and apoptosis.

The team uses advanced techniques like the Orbitrap Eclipse mass spectrometer and the SLN1 single-cell isolation system to isolate and analyze individual cells. This approach allows them to observe the upregulation of proteins involved in DNA repair and the arrest of cells in the S phase of the cell cycle. The findings highlight the importance of single-cell proteomics in revealing cellular heterogeneity and the dynamic response to DNA damage, which could have implications for understanding diseases like cancer and neurodegenerative disorders. 

Learning points:

• Single-Cell Proteomics Advantage: Single-cell proteomics allows for the observation of individual cellular responses to DNA damage, revealing heterogeneity that would be masked in bulk proteomics. This approach helps in understanding the specific activation of DNA repair mechanisms and cell cycle arrest at the S phase. 
• DNA Repair Mechanisms: The study highlights the activation of various DNA repair pathways, such as mismatch repair, base excision repair, and nucleotide excision repair, in response to DNA damage induced by the compound 60G. This understanding is crucial for insights into diseases like cancer and neurodegenerative disorders. 
• Technological Integration: The use of advanced technologies like the Orbitrap Eclipse mass spectrometer and the SLN1 single-cell isolation system enables precise isolation and analysis of single cells, facilitating detailed proteomic studies and enhancing the understanding of cellular processes at a granular level. 

Who should attend:

• Cancer Researchers: Understanding the variability in DNA repair mechanisms at the single-cell level can provide insights into tumor heterogeneity and the development of targeted cancer therapies.
• Neurodegenerative Disease Scientists: Insights into DNA damage response mechanisms can aid in understanding the molecular basis of neurodegenerative diseases and potentially lead to new therapeutic strategies. 
• Proteomics and Genomics Technologists: Professionals working with advanced proteomics and genomics technologies can benefit from the presentation's focus on cutting-edge techniques for single-cell analysis, enhancing their research capabilities and methodologies. 

If you cannot attend this webinar please register to receive a link to the On Demand version the following day. 

Presenter: Prof. Konstantinos Thalassinos (Professor, University College London)

Kostas moved to the UK in 1998 to study for a BSc in Genetics at the University of Leicester, having completed his high school education in his native Greece. After graduation he went on to the University of York to undertake a Masters in Bioinformatics, which included a three-month industrial placement writing software to process mass spectrometric data. Having gained an interest in mass spectrometry, Kostas then went to the University of Warwick in order to pursue a PhD that combined experimental and computational studies in mass spectrometry-based proteomics. After being awarded his PhD in 2006, Kostas was the successful recipient of a one-year Wellcome Trust Value in People Fellowship. Post-doctoral training followed and in 2010 he moved to the Institute of Structural and Molecular Biology at University College London / Birkbeck to take up a lectureship in biophysical mass spectrometry and has been there ever since. In 2015 he was promoted to Senior Lecturer and in 2019 to Professor of mass spectrometry.

His lab is using structural mass spectrometry approaches, especially ion mobility and crosslinking, to study the structure and dynamics of proteins and protein complexes, particularly those involved in protein misfolding diseases.