Abstract
We describe a multiplexed imaging mass spectrometry approach especially suitable for fibrosis research. Fibrosis is a process characterized by excessive extracellular matrix (ECM) secretion. Buildup of ECM impairs tissue and organ function to promote further progression of disease. It is an ongoing analytical challenge to access ECM for diagnosis and therapeutic treatment of fibrosis. Recently, we reported the use of the enzyme collagenase type III to target the ECM proteome in thin histological tissue sections of fibrotic diseases including hepatocellular carcinoma, breast cancer, prostate cancer, lung cancer and aortic valve stenosis. Detection of collagenase type III peptides by matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) allows localization of ECM peptide sequences to specific regions of fibrosis. We have further identified that the ECM proteome accessed by collagenase type III has on average 3.7 sites per protein that may be differentially N-glycosylated. N-glycosylation is a major posttranslational modification of the ECM proteome, influencing protein folding, secretion, and organization. Understanding both N-glycosylation signaling and regulation of ECM expression significantly informs on ECM signaling in fibrosis.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cox TR, Erler JT (2011) Remodeling and homeostasis of the extracellular matrix: implications for fibrotic diseases and cancer. Dis Model Mech 4:165–178. https://doi.org/10.1242/dmm.004077
Fan D, Takawale A, Lee J, Kassiri Z (2012) Cardiac fibroblasts, fibrosis and extracellular matrix remodeling in heart disease. Fibrogenes Tissue Repair 5(1):15. https://doi.org/10.1186/1755-1536-5-15
Walraven M, Hinz B (2018) Therapeutic approaches to control tissue repair and fibrosis: extracellular matrix as a game changer. Matrix Biol 71-72:205–224. https://doi.org/10.1016/j.matbio.2018.02.020
Schwamborn K, Kriegsmann M, Weichert W (2017) MALDI imaging mass spectrometry — from bench to bedside. Biochim Biophys Acta Proteins Proteom 1865(7):776–783. https://doi.org/10.1016/j.bbapap.2016.10.014
West CA, Wang M, Herrera H et al (2018) N-linked glycan branching and fucosylation are increased directly in Hcc tissue as determined through in situ glycan imaging. J Proteome Res 17:3454–3462. https://doi.org/10.1021/acs.jproteome.8b00323
Drake RR, West CA, Mehta AS, Angel PM (2018) MALDI mass spectrometry imaging of N-linked glycans in tissues. Springer, Singapore, pp 59–76
Powers T, Holst S, Wuhrer M et al (2015) Two-dimensional N-glycan distribution mapping of hepatocellular carcinoma tissues by MALDI-imaging mass spectrometry. Biomol Ther 5:2554–2572. https://doi.org/10.3390/biom5042554
Powers TW, Jones EE, Betesh LR et al (2013) Matrix assisted laser desorption ionization imaging mass spectrometry workflow for spatial profiling analysis of N-linked glycan expression in tissues. Anal Chem 85:9799–9806. https://doi.org/10.1021/ac402108x
Angel PM, Mehta A, Norris-Caneda K, Drake RR (2018) MALDI imaging mass spectrometry of N-glycans and tryptic peptides from the same formalin-fixed, paraffin-embedded tissue section. Methods Mol Biol 1788:225–241. https://doi.org/10.1007/7651_2017_81
Angel PM, Bayoumi AS, Hinton RB et al (2016) MALDI-IMS as a lipidomic approach to heart valve research. J Heart Valve Dis 25:240–252
Scott DA, Casadonte R, Cardinali B et al (2019) Increases in tumor N-glycan polylactosamines associated with advanced HER2-positive and triple-negative breast Cancer tissues. Proteomics Clin Appl 13(1):e1800014. https://doi.org/10.1002/prca.201800014
Powers TW, Neely BA, Shao Y et al (2014) MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. PLoS One 9(9):e106255. https://doi.org/10.1371/journal.pone.0106255
Angel, PM (Corresponding author), Comte-Walters, Ball, LE, Talbot, K, Mehta AS, Drake RR (2018) Mapping extracellular matrix proteins in formalin-Fixed, paraffin-embedded tissues by MALDI imaging mass spectrometry.J Proteome Res 17:635–646. https://doi.org/10.1021/acs.jproteome.7b00713
Angel, PM (Corresponding author), Bruner E, Bethard JR, Clift CL, Ball LE, Drake RR, Feghali-Bostwick C (2019) Extracellular matrix alterations in low grade lung adenocarcinoma compared to normal lung tissue by imaging mass spectrometry. J Mass Spect 55(4):e4450. https://doi.org/10.1002/jms.4450
Ceroni A, Maass K, Geyer H et al (2008) GlycoWorkbench: a tool for the computer-assisted annotation of mass spectra of glycans. J Proteome Res 7:1650–1659. https://doi.org/10.1021/pr7008252
Acknowledgments
PMA and CLC are supported by P20GM103542 (NIH/NIGMS), HL007260 (NHLBI), and in part by pilot research funding, Hollings Cancer Center’s Cancer Center Support Grant P30 CA138313 at the Medical University of South Carolina. Additional support was provided by the South Carolina Centers of Economic Excellence SmartState program to RRD and ASM. The MUSC Mass Spectrometry Facility is supported by the Office of the Provost and the South Carolina COBRE in Oxidants, Redox Balance and Stress Signaling (P20GM 103542).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Clift, C.L., Mehta, A., Drake, R.R., Angel, P.M. (2021). Multiplexed Imaging Mass Spectrometry of Histological Staining, N-Glycan and Extracellular Matrix from One Tissue Section: A Tool for Fibrosis Research. In: Zamir, E. (eds) Multiplexed Imaging. Methods in Molecular Biology, vol 2350. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1593-5_20
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1593-5_20
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1592-8
Online ISBN: 978-1-0716-1593-5
eBook Packages: Springer Protocols