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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...
Autores principales: | , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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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. |
format | Online Article Text |
id | pubmed-9709465 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
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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