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An optimized approach and inflation media for obtaining complimentary mass spectrometry-based omics data from human lung tissue

Human disease states are biomolecularly multifaceted and can span across phenotypic states, therefore it is important to understand diseases on all levels, across cell types, and within and across microanatomical tissue compartments. To obtain an accurate and representative view of the molecular lan...

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Autores principales: Lukowski, Jessica K., Olson, Heather, Velickovic, Marija, Wang, Juan, Kyle, Jennifer E., Kim, Young-Mo, Williams, Sarah M., Zhu, Ying, Huyck, Heidi L., McGraw, Matthew D., Poole, Cory, Rogers, Lisa, Misra, Ravi, Alexandrov, Theodore, Ansong, Charles, Pryhuber, Gloria S., Clair, Geremy, Adkins, Joshua N., Carson, James P., Anderton, Christopher R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9709465/
https://www.ncbi.nlm.nih.gov/pubmed/36465564
http://dx.doi.org/10.3389/fmolb.2022.1022775
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author Lukowski, Jessica K.
Olson, Heather
Velickovic, Marija
Wang, Juan
Kyle, Jennifer E.
Kim, Young-Mo
Williams, Sarah M.
Zhu, Ying
Huyck, Heidi L.
McGraw, Matthew D.
Poole, Cory
Rogers, Lisa
Misra, Ravi
Alexandrov, Theodore
Ansong, Charles
Pryhuber, Gloria S.
Clair, Geremy
Adkins, Joshua N.
Carson, James P.
Anderton, Christopher R.
author_facet Lukowski, Jessica K.
Olson, Heather
Velickovic, Marija
Wang, Juan
Kyle, Jennifer E.
Kim, Young-Mo
Williams, Sarah M.
Zhu, Ying
Huyck, Heidi L.
McGraw, Matthew D.
Poole, Cory
Rogers, Lisa
Misra, Ravi
Alexandrov, Theodore
Ansong, Charles
Pryhuber, Gloria S.
Clair, Geremy
Adkins, Joshua N.
Carson, James P.
Anderton, Christopher R.
author_sort Lukowski, Jessica K.
collection PubMed
description Human disease states are biomolecularly multifaceted and can span across phenotypic states, therefore it is important to understand diseases on all levels, across cell types, and within and across microanatomical tissue compartments. To obtain an accurate and representative view of the molecular landscape within human lungs, this fragile tissue must be inflated and embedded to maintain spatial fidelity of the location of molecules and minimize molecular degradation for molecular imaging experiments. Here, we evaluated agarose inflation and carboxymethyl cellulose embedding media and determined effective tissue preparation protocols for performing bulk and spatial mass spectrometry-based omics measurements. Mass spectrometry imaging methods were optimized to boost the number of annotatable molecules in agarose inflated lung samples. This optimized protocol permitted the observation of unique lipid distributions within several airway regions in the lung tissue block. Laser capture microdissection of these airway regions followed by high-resolution proteomic analysis allowed us to begin linking the lipidome with the proteome in a spatially resolved manner, where we observed proteins with high abundance specifically localized to the airway regions. We also compared our mass spectrometry results to lung tissue samples preserved using two other inflation/embedding media, but we identified several pitfalls with the sample preparation steps using this preservation method. Overall, we demonstrated the versatility of the inflation method, and we can start to reveal how the metabolome, lipidome, and proteome are connected spatially in human lungs and across disease states through a variety of different experiments.
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spelling pubmed-97094652022-12-01 An optimized approach and inflation media for obtaining complimentary mass spectrometry-based omics data from human lung tissue Lukowski, Jessica K. Olson, Heather Velickovic, Marija Wang, Juan Kyle, Jennifer E. Kim, Young-Mo Williams, Sarah M. Zhu, Ying Huyck, Heidi L. McGraw, Matthew D. Poole, Cory Rogers, Lisa Misra, Ravi Alexandrov, Theodore Ansong, Charles Pryhuber, Gloria S. Clair, Geremy Adkins, Joshua N. Carson, James P. Anderton, Christopher R. Front Mol Biosci Molecular Biosciences Human disease states are biomolecularly multifaceted and can span across phenotypic states, therefore it is important to understand diseases on all levels, across cell types, and within and across microanatomical tissue compartments. To obtain an accurate and representative view of the molecular landscape within human lungs, this fragile tissue must be inflated and embedded to maintain spatial fidelity of the location of molecules and minimize molecular degradation for molecular imaging experiments. Here, we evaluated agarose inflation and carboxymethyl cellulose embedding media and determined effective tissue preparation protocols for performing bulk and spatial mass spectrometry-based omics measurements. Mass spectrometry imaging methods were optimized to boost the number of annotatable molecules in agarose inflated lung samples. This optimized protocol permitted the observation of unique lipid distributions within several airway regions in the lung tissue block. Laser capture microdissection of these airway regions followed by high-resolution proteomic analysis allowed us to begin linking the lipidome with the proteome in a spatially resolved manner, where we observed proteins with high abundance specifically localized to the airway regions. We also compared our mass spectrometry results to lung tissue samples preserved using two other inflation/embedding media, but we identified several pitfalls with the sample preparation steps using this preservation method. Overall, we demonstrated the versatility of the inflation method, and we can start to reveal how the metabolome, lipidome, and proteome are connected spatially in human lungs and across disease states through a variety of different experiments. Frontiers Media S.A. 2022-11-16 /pmc/articles/PMC9709465/ /pubmed/36465564 http://dx.doi.org/10.3389/fmolb.2022.1022775 Text en Copyright © 2022 Lukowski, Olson, Velickovic, Wang, Kyle, Kim, Williams, Zhu, Huyck, McGraw, Poole, Rogers, Misra, Alexandrov, Ansong, Pryhuber, Clair, Adkins, Carson and Anderton. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Molecular Biosciences
Lukowski, Jessica K.
Olson, Heather
Velickovic, Marija
Wang, Juan
Kyle, Jennifer E.
Kim, Young-Mo
Williams, Sarah M.
Zhu, Ying
Huyck, Heidi L.
McGraw, Matthew D.
Poole, Cory
Rogers, Lisa
Misra, Ravi
Alexandrov, Theodore
Ansong, Charles
Pryhuber, Gloria S.
Clair, Geremy
Adkins, Joshua N.
Carson, James P.
Anderton, Christopher R.
An optimized approach and inflation media for obtaining complimentary mass spectrometry-based omics data from human lung tissue
title An optimized approach and inflation media for obtaining complimentary mass spectrometry-based omics data from human lung tissue
title_full An optimized approach and inflation media for obtaining complimentary mass spectrometry-based omics data from human lung tissue
title_fullStr An optimized approach and inflation media for obtaining complimentary mass spectrometry-based omics data from human lung tissue
title_full_unstemmed An optimized approach and inflation media for obtaining complimentary mass spectrometry-based omics data from human lung tissue
title_short An optimized approach and inflation media for obtaining complimentary mass spectrometry-based omics data from human lung tissue
title_sort optimized approach and inflation media for obtaining complimentary mass spectrometry-based omics data from human lung tissue
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9709465/
https://www.ncbi.nlm.nih.gov/pubmed/36465564
http://dx.doi.org/10.3389/fmolb.2022.1022775
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