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

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.

journal_name

J Biomol NMR

journal_title

Journal of biomolecular NMR

authors

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

doi

10.1007/s10858-020-00328-8

subject

Has Abstract

pub_date

2020-09-01 00:00:00

pages

401-412

issue

8-9

eissn

0925-2738

issn

1573-5001

pii

10.1007/s10858-020-00328-8

journal_volume

74

pub_type

杂志文章

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