Adrenal Hormone Interactions and Metabolism: A Single Sample
                    Multi-Omics Approach

Nicole Bechmann; Deepika Watts; Charlotte Steenblock; Paal William Wallace; Annette Schürmann; Stefan R. Bornstein; Ben Wielockx; Graeme Eisenhofer; Mirko Peitzsch

doi:10.1055/a-1440-0278

Hormone and Metabolic Research, Inhaltsverzeichnis

CC BY-NC-ND 4.0 · Horm Metab Res 2021; 53(05): 326-334
DOI: 10.1055/a-1440-0278

Endocrine Care

Adrenal Hormone Interactions and Metabolism: A Single Sample Multi-Omics Approach

Nicole Bechmann

¹Institute of Clinical Chemistry and Laboratory Medicine, Technische Universität Dresden, Dresden, Germany

²Department of Medicine III, Technische Universität Dresden, Dresden, Germany

³German Institute of Human Nutrition Potsdam-Rehbruecke, Department of Experimental Diabetology, Nuthetal, Germany

⁴German Center for Diabetes Research (DZD), München-Neuherberg, Germany

,

Deepika Watts

¹Institute of Clinical Chemistry and Laboratory Medicine, Technische Universität Dresden, Dresden, Germany

,

Charlotte Steenblock

²Department of Medicine III, Technische Universität Dresden, Dresden, Germany

,

Paal William Wallace

¹Institute of Clinical Chemistry and Laboratory Medicine, Technische Universität Dresden, Dresden, Germany

,

Annette Schürmann

³German Institute of Human Nutrition Potsdam-Rehbruecke, Department of Experimental Diabetology, Nuthetal, Germany

⁴German Center for Diabetes Research (DZD), München-Neuherberg, Germany

,

Stefan R. Bornstein

²Department of Medicine III, Technische Universität Dresden, Dresden, Germany

,

Ben Wielockx

¹Institute of Clinical Chemistry and Laboratory Medicine, Technische Universität Dresden, Dresden, Germany

,

Graeme Eisenhofer

¹Institute of Clinical Chemistry and Laboratory Medicine, Technische Universität Dresden, Dresden, Germany

²Department of Medicine III, Technische Universität Dresden, Dresden, Germany

,

Mirko Peitzsch

¹Institute of Clinical Chemistry and Laboratory Medicine, Technische Universität Dresden, Dresden, Germany

› Institutsangaben

Abstract

The adrenal gland is important for many physiological and pathophysiological processes, but studies are often restricted by limited availability of sample material. Improved methods for sample preparation are needed to facilitate analyses of multiple classes of adrenal metabolites and macromolecules in a single sample. A procedure was developed for preparation of chromaffin cells, mouse adrenals, and human chromaffin tumors that allows for multi-omics analyses of different metabolites and preservation of native proteins. To evaluate the new procedure, aliquots of samples were also prepared using conventional procedures. Metabolites were analyzed by liquid-chromatography with mass spectrometry or electrochemical detection. Metabolite contents of chromaffin cells and tissues analyzed with the new procedure were similar or even higher than with conventional methods. Catecholamine contents were comparable between both procedures. The TCA cycle metabolites, cis-aconitate, isocitate, and α-ketoglutarate were detected at higher concentrations in cells, while in tumor tissue only isocitrate and potentially fumarate were measured at higher contents. In contrast, in a broad untargeted metabolomics approach, a methanol-based preparation procedure of adrenals led to a 1.3-fold higher number of detected metabolites. The established procedure also allows for simultaneous investigation of adrenal hormones and related enzyme activities as well as proteins within a single sample. This novel multi-omics approach not only minimizes the amount of sample required and overcomes problems associated with tissue heterogeneity, but also provides a more complete picture of adrenal function and intra-adrenal interactions than previously possible.

Key words

adrenal - steroids - catecholamines - TCA-cycle metabolites - preservation of native proteins - cortical-medullary interaction

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Referenzen

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