Spatial Biology Europe

Join leaders, experts, and researchers at the Spatial Biology Europe: In-Person, connecting global academic & research organizations as well as pharma representatives for high-level discussions on the latest innovations in spatial research & technologies. The Yokogawa Life Science team Europe will showcase our Single Cellome Systems. 

   On June 8th, Ph.D. Carla Newman (Associate Director GSK) will hold a talk about "SiLC-MS (Single Live-Cell Mass Spectrometry) analysis in the context of drug discovery".     

   

 Abstract

In the last few decades, the pharmaceutical industry has transformed people’s lives. However, the development of new drugs is becoming increasingly difficult and a paradigm shift in the drug discovery workflow is required to reduce attrition and transform conventional drug screening assays into translatable analytical techniques for the analysis of drugs in complex environments, both in-vitro and ex-vivo. The ability to visualise unlabelled compounds inside the cell at physiological dosages can offer valuable insight into the compound behaviour both on and off-target.

SiLC-MS is a semi-automated methodology that allows the collection of intracellular contents using a modified CQ1 imaging system developed by Yokowaga. The instrument is equipped with a confocal microscope that allows bright field imaging as well as fluorescence imaging with 4 lasers (405, 488, 561 and 640 nm). Sampling is performed using the tips developed by Professor Masujima (1-4). The tip, holding the cellular contents, is then used for static nanospray of the contents into an Orbitrap Fusion Lumos (Thermo Scientific) and the resulting data processed using Compound Discoverer (Thermo Scientific).

In this study, we show the applicability of the SiLC-MS technology to drug discovery, as it is crucial to identify compound and its metabolites when incubated in a mammalian cell at a therapeutic dose. We report on the validation studies performed using the SiLC-MS platform, in these validation studies we assess the ability to distinguish different cell types based on their metabolomic fingerprint, furthermore, we have also evaluated if this assay was sensitive enough to detect drugs intracellularly.

 

We are currently establishing a multi-omics platform on the modified CQ1 that allows both metabolomics and transcriptomics at the single-cell level. For that, we have sampled the cells first for metabolomics and then for transcriptomics.

 

We demonstrate that dosed compound can be identified in a single cell after sampling using the modified CQ1, endogenous metabolites can also be identified that can further the understanding of the drug’s mechanism. This technique has direct relevance for assessing compound effects on disease-relevant cells and its low sample requirement makes it applicable to studying rare cell types. The use of high content imaging system enables the effect of compounds on live cells to be studied and suitable time points selected for sampling cell contents.

 

References:

1) Fujii T., Matsuda S., Tejedor M.J.,Esaki T., Sakane I., Mizuno H., Tsuyama N.,  Masujima T.,

Nature Protocols volume 10, pages 1445–1456 (2015) doi:10.1038/nprot.2015.084

2) Masujima T. Anal Sci. 2009 Aug;25(8):953-60.

3) Mizuno H., Tsuyama N., Harada T., Masujima T., J Mass Spectrom. 2008 Dec;43(12):1692-700. doi: 10.1002/jms.1460.

4) Guillaume-Gentil O., Rey O., Kiefer P., Ibáñez A.J., Steinhoff R., Brönnimann R., Dorwling-Carter L., Zambelli T., Zenobi R. , Vorholt J.A.,  Anal. Chem., 2017, 89 (9), pp 5017–5023 DOI: 10.1021/acs.analchem.7b00367