Development of in vitro-grown spheroids as a 3D tumor model system for solid-state NMR spectroscopy.
 

Development of in vitro-grown spheroids as a 3D tumor model system for solid-state NMR spectroscopy.

http://www.bionmr.com//www.ncbi.nlm....ringerlink.gif Related Articles Development of in vitro-grown spheroids as a 3D tumor model system for solid-state NMR spectroscopy.

J Biomol NMR. 2020 Jun 19;:

Authors: Damman R, Lucini Paioni A, Xenaki KT, Beltrán Hernández I, van Bergen En Henegouwen PMP, Baldus M

Abstract
Recent advances in the field of in-cell NMR spectroscopy have made it possible to study proteins in the context of bacterial or mammalian cell extracts or even entire cells. As most mammalian cells are part of a multi-cellular complex, there is a need to develop novel NMR approaches enabling the study of proteins within the complexity of a 3D cellular environment. Here we investigate the use of the hanging drop method to grow spheroids which are homogenous in size and shape as a model system to study solid tumors using solid-state NMR (ssNMR) spectroscopy. We find that these spheroids are stable under magic-angle-spinning conditions and show a clear change in metabolic profile as compared to single cell preparations. Finally, we utilize dynamic nuclear polarization (DNP)-supported ssNMR measurements to show that low concentrations of labelled nanobodies targeting EGFR (7D12) can be detected inside the spheroids. These findings suggest that solid-state NMR can be used to directly examine proteins or other biomolecules in a 3D cellular microenvironment with potential applications in pharmacological research.


PMID: 32562030 [PubMed - as supplied by publisher]



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