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Mapping the triglyceride distribution in NAFLD human liver by MALDI imaging mass spectrometry reveals molecular differences in micro and macro steatosis

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Abstract

Hepatic lipid accumulation, mainly in the form of triglycerides (TGs), is the hallmark of non-alcoholic fatty liver disease (NAFLD). To date, the spatial distribution of individual lipids in NAFLD-affected livers is not well characterized. This study aims to map the triglyceride distribution in normal human liver samples and livers with NAFLD and cirrhosis with imaging mass spectrometry (MALDI IMS). Specifically, whether individual triglyceride species differing by fatty acid chain length and degree of saturation correlate with the histopathological features of NAFLD as identified with classical H&E. Using a recently reported sodium-doped gold-assisted laser desorption/ionization IMS sample preparation, 20 human liver samples (five normal livers, five samples with simple steatosis, five samples with steatohepatitis, and five samples with cirrhosis) were analyzed at 10-μm lateral resolution. A total of 24 individual lipid species, primarily neutral lipids, were identified (22 TGs and two phospholipids). In samples with a low level of steatosis, TGs accumulated around the pericentral zone. In all samples, TGs with different degrees of side-chain saturation and side-chain length demonstrated differential distribution. Furthermore, hepatocytes containing macro lipid droplets were highly enriched in fully saturated triglycerides. This enrichment was also observed in areas of hepatocyte ballooning in samples with steatohepatitis and cirrhosis. In conclusion, macro lipid droplets in NAFLD are enriched in fully saturated triglycerides, indicating a possible increase in de novo lipogenesis that leads to steatohepatitis and cirrhosis.

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Abbreviations

AuLDI IMS:

Gold-assisted laser desorption/ionization imaging mass spectrometry

DAG:

Diglyceride

DNL:

De novo lipogenesis

FFAs:

Free fatty acids

MALDI IMS:

Matrix-assisted laser desorption/ionization imaging mass spectrometry

NAFLD:

Non-alcoholic fatty liver disease

NASH:

Non-alcoholic steatohepatitis

OCT:

Optimal cutting temperature compound

SFAs:

Saturated fatty acids

SS:

Simple steatosis

TG:

Triglycerides

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Acknowledgments

We would like to acknowledge the support of Stephanie Petrillo and Abdellatif Amri for technical support, the MUHC Liver Disease Biobank for all human specimens, and the patients who consented to providing samples to the Biobank, with whose support of this study would not have been possible.

Funding

This publication was supported by National Cancer Institute of the National Institutes of Health under award number R01CA198103 and by the Natural Sciences and Engineering Research Council of Canada.

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Author notes

  1. Nathan Heath Patterson

    Present address: Mass Spectrometry Research Center, Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, 37240, USA

Authors and Affiliations

  1. Department of Surgery, McGill University Health Center, 1650 Cedar Avenue, Montreal, Quebec, H3G 1A4, Canada

    Hussam Alamri & Peter Metrakos

  2. Department of Chemistry, University of Montreal, Montreal, Quebec, H3C 3J7, Canada

    Nathan Heath Patterson, Ethan Yang & Pierre Chaurand

  3. Pathology Department, School of Medicine, Pontificia Universidad Católica de Chile, Avda. Libertador Bernardo O’Higgins 340, 8331150, Santiago, Chile

    Pablo Zoroquiain

  4. Cancer Research Program, McGill University Health Center-Research Institute, Montreal, Quebec, H4A 3J1, Canada

    Anthoula Lazaris & Peter Metrakos

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Corresponding authors

Correspondence to Pierre Chaurand or Peter Metrakos.

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Informed consent was obtained from all patients through the McGill University Health Center Liver Disease Biobank. The study protocol conformed to the ethical guidelines of the 1975 Declaration of Helsinki and was approved by the research ethics board of the McGill University Health Centre.

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Alamri, H., Patterson, N.H., Yang, E. et al. Mapping the triglyceride distribution in NAFLD human liver by MALDI imaging mass spectrometry reveals molecular differences in micro and macro steatosis. Anal Bioanal Chem 411, 885–894 (2019). https://doi.org/10.1007/s00216-018-1506-8

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