Spatial quantitative metabolomics enables identification of remote and sustained ipsilateral cortical metabolic reprogramming after stroke

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

Abstract Mass spectrometry imaging (MSI) has become a cornerstone of spatial biology research. However, various factors that are intrinsic to the technology limit quantitative capacity of MSI-based spatial metabolomics and thus reliable interpretation. Here, we developed a quantitative MSI workflow (Q-MSI), based on isotopically 13 C-labeled yeast extract as internal standards, to overcome these pitfalls. Using brain and kidney tissue, we demonstrate that this approach allows for (absolute) quantification of hundreds of metabolites and lipids. Applying our workflow to a stroke model allowed us to not only map metabolic remodeling of the infarct and peri-infarct area over time, but also discover hitherto unnoted remote metabolic remodeling in the histologically unaffected ipsilateral sensorimotor cortex. At day 7 post-stroke, increased levels of neuroprotective lysine and reduced excitatory glutamate levels were found compared to the contralateral cortex. By day 28 post-stroke, lysine and glutamate levels had been normalized, while decreased precursor pools of UDP-GlcNAc and linoleate persisted that have previously been associated with vulnerability. Importantly, traditional normalization strategies not employing internal standards were unable to visualize these differences. Using 13 C-labeled yeast extracts as a normalization strategy establishes a new paradigm in quantitative MSI-based spatial metabolomics that greatly enhances reliability and interpretive strength.

Authors 8

1. Gangqi Wang corresponding Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine

   Leiden University · Fudan University · Leiden University Medical Center · Children's Hospital of Fudan University

   Affiliation as printed

   Children’s Hospital of Fudan University, and the Shanghai Key Laboratory of Medical Epigenetics, Institutes of Biomedical Sciences, Fudan University, Shanghai, China

   Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, The Netherlands

2. Bernard M. van den Berg Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)

   Leiden University · Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine

   Affiliation as printed

   Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, The Netherlands

   The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, Leiden, The Netherlands

3. Sarantos Kostidis Aachen Center of Proteomics and Metabolomics

   Leiden University Medical Center

   Affiliation as printed

   Center of Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands

4. Kelsey Pinkham

   LMU Klinikum · Munich Cluster for Systems Neurology · Institut für Schlaganfall- und Demenzforschung · Ludwig-Maximilians-Universität München

   Affiliation as printed

   Institute for Stroke and Dementia Research (ISD), University Hospital, LMU Munich, Munich, Germany; Munich Cluster for Systems Neurology (SyNergy), Munich, Germany

5. Marleen E Jacobs Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)

   Leiden University · Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine

   Affiliation as printed

   Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, The Netherlands

   The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, Leiden, The Netherlands

6. Arthur Liesz

   LMU Klinikum · Munich Cluster for Systems Neurology · Institut für Schlaganfall- und Demenzforschung · Ludwig-Maximilians-Universität München

   Affiliation as printed

   Institute for Stroke and Dementia Research (ISD), University Hospital, LMU Munich, Munich, Germany; Munich Cluster for Systems Neurology (SyNergy), Munich, Germany

7. Martin A. Giera corresponding Aachen Center of Proteomics and Metabolomics Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)

   Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine

   Affiliation as printed

   Center of Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands

   The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, Leiden, The Netherlands

8. Ton J. Rabelink corresponding Aachen Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)

   Leiden University · Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine

   Affiliation as printed

   Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, The Netherlands

   The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, Leiden, The Netherlands