Ulcerative Colitis-Derived Colonoid Culture: A Multi-Mineral-Approach to Improve Barrier Protein Expression
BackgroundRecent studies demonstrated that Aquamin®, a calcium-, magnesium-rich, multi-mineral natural product, improves barrier structure and function in colonoids obtained from the tissue of healthy subjects. The goal of the present study was to determine if the colonic barrier could be improved i...
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| Format: | Article |
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Frontiers Media S.A.
2020-11-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2020.577221/full |
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| author | Muhammad N. Aslam Shannon D. McClintock Durga Attili Shailja Pandya Humza Rehman Daniyal M. Nadeem Mohamed Ali H. Jawad-Makki Areeba H. Rizvi Maliha M. Berner Michael K. Dame Danielle Kim Turgeon James Varani |
| author_facet | Muhammad N. Aslam Shannon D. McClintock Durga Attili Shailja Pandya Humza Rehman Daniyal M. Nadeem Mohamed Ali H. Jawad-Makki Areeba H. Rizvi Maliha M. Berner Michael K. Dame Danielle Kim Turgeon James Varani |
| author_sort | Muhammad N. Aslam |
| collection | DOAJ |
| description | BackgroundRecent studies demonstrated that Aquamin®, a calcium-, magnesium-rich, multi-mineral natural product, improves barrier structure and function in colonoids obtained from the tissue of healthy subjects. The goal of the present study was to determine if the colonic barrier could be improved in tissue from subjects with ulcerative colitis (UC).MethodsColonoid cultures were established with colon biopsies from 9 individuals with UC. The colonoids were then incubated for a 2-week period under control conditions (in culture medium with a final calcium concentration of 0.25 mM) or in the same medium supplemented with Aquamin® to provide 1.5 – 4.5 mM calcium. Effects on differentiation and barrier protein expression were determined using several approaches: phase-contrast and scanning electron microscopy, quantitative histology and immunohistology, mass spectrometry-based proteome assessment and transmission electron microscopy.ResultsAlthough there were no gross changes in colonoid appearance, there was an increase in lumen diameter and wall thickness on histology and greater expression of cytokeratin 20 (CK20) along with reduced expression of Ki67 by quantitative immunohistology observed with intervention. In parallel, upregulation of several differentiation-related proteins was seen in a proteomic screen with the intervention. Aquamin®-treated colonoids demonstrated a modest up-regulation of tight junctional proteins but stronger induction of adherens junction and desmosomal proteins. Increased desmosomes were seen at the ultrastructural level. Proteomic analysis demonstrated increased expression of several basement membrane proteins and hemidesmosomal components. Proteins expressed at the apical surface (mucins and trefoils) were also increased as were several additional proteins with anti-microbial activity or that modulate inflammation. Finally, several transporter proteins that affect electrolyte balance (and, thereby affect water resorption) were increased. At the same time, growth and cell cycle regulatory proteins (Ki67, nucleophosmin, and stathmin) were significantly down-regulated. Laminin interactions, matrix formation and extracellular matrix organization were the top three up-regulated pathways with the intervention.ConclusionA majority of individuals including patients with UC do not reach the recommended daily intake for calcium and other minerals. To the extent that such deficiencies might contribute to the weakening of the colonic barrier, the findings employing UC tissue-derived colonoids here suggest that adequate mineral intake might improve the colonic barrier. |
| first_indexed | 2024-12-11T16:12:55Z |
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| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-12-11T16:12:55Z |
| publishDate | 2020-11-01 |
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| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-c1644ff21bb74747ac2011092a6e57712022-12-22T00:59:02ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2020-11-01810.3389/fcell.2020.577221577221Ulcerative Colitis-Derived Colonoid Culture: A Multi-Mineral-Approach to Improve Barrier Protein ExpressionMuhammad N. Aslam0Shannon D. McClintock1Durga Attili2Shailja Pandya3Humza Rehman4Daniyal M. Nadeem5Mohamed Ali H. Jawad-Makki6Areeba H. Rizvi7Maliha M. Berner8Michael K. Dame9Danielle Kim Turgeon10James Varani11Department of Pathology, The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Pathology, The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Pathology, The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Pathology, The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Pathology, The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Pathology, The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Pathology, The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Pathology, The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Internal Medicine (The Division of Gastroenterology), The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Internal Medicine (The Division of Gastroenterology), The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Internal Medicine (The Division of Gastroenterology), The University of Michigan Medical School, Ann Arbor, MI, United StatesDepartment of Pathology, The University of Michigan Medical School, Ann Arbor, MI, United StatesBackgroundRecent studies demonstrated that Aquamin®, a calcium-, magnesium-rich, multi-mineral natural product, improves barrier structure and function in colonoids obtained from the tissue of healthy subjects. The goal of the present study was to determine if the colonic barrier could be improved in tissue from subjects with ulcerative colitis (UC).MethodsColonoid cultures were established with colon biopsies from 9 individuals with UC. The colonoids were then incubated for a 2-week period under control conditions (in culture medium with a final calcium concentration of 0.25 mM) or in the same medium supplemented with Aquamin® to provide 1.5 – 4.5 mM calcium. Effects on differentiation and barrier protein expression were determined using several approaches: phase-contrast and scanning electron microscopy, quantitative histology and immunohistology, mass spectrometry-based proteome assessment and transmission electron microscopy.ResultsAlthough there were no gross changes in colonoid appearance, there was an increase in lumen diameter and wall thickness on histology and greater expression of cytokeratin 20 (CK20) along with reduced expression of Ki67 by quantitative immunohistology observed with intervention. In parallel, upregulation of several differentiation-related proteins was seen in a proteomic screen with the intervention. Aquamin®-treated colonoids demonstrated a modest up-regulation of tight junctional proteins but stronger induction of adherens junction and desmosomal proteins. Increased desmosomes were seen at the ultrastructural level. Proteomic analysis demonstrated increased expression of several basement membrane proteins and hemidesmosomal components. Proteins expressed at the apical surface (mucins and trefoils) were also increased as were several additional proteins with anti-microbial activity or that modulate inflammation. Finally, several transporter proteins that affect electrolyte balance (and, thereby affect water resorption) were increased. At the same time, growth and cell cycle regulatory proteins (Ki67, nucleophosmin, and stathmin) were significantly down-regulated. Laminin interactions, matrix formation and extracellular matrix organization were the top three up-regulated pathways with the intervention.ConclusionA majority of individuals including patients with UC do not reach the recommended daily intake for calcium and other minerals. To the extent that such deficiencies might contribute to the weakening of the colonic barrier, the findings employing UC tissue-derived colonoids here suggest that adequate mineral intake might improve the colonic barrier.https://www.frontiersin.org/articles/10.3389/fcell.2020.577221/fullbasement membranecalciumcell barriercolonoiddesmosomeorganoid culture |
| spellingShingle | Muhammad N. Aslam Shannon D. McClintock Durga Attili Shailja Pandya Humza Rehman Daniyal M. Nadeem Mohamed Ali H. Jawad-Makki Areeba H. Rizvi Maliha M. Berner Michael K. Dame Danielle Kim Turgeon James Varani Ulcerative Colitis-Derived Colonoid Culture: A Multi-Mineral-Approach to Improve Barrier Protein Expression Frontiers in Cell and Developmental Biology basement membrane calcium cell barrier colonoid desmosome organoid culture |
| title | Ulcerative Colitis-Derived Colonoid Culture: A Multi-Mineral-Approach to Improve Barrier Protein Expression |
| title_full | Ulcerative Colitis-Derived Colonoid Culture: A Multi-Mineral-Approach to Improve Barrier Protein Expression |
| title_fullStr | Ulcerative Colitis-Derived Colonoid Culture: A Multi-Mineral-Approach to Improve Barrier Protein Expression |
| title_full_unstemmed | Ulcerative Colitis-Derived Colonoid Culture: A Multi-Mineral-Approach to Improve Barrier Protein Expression |
| title_short | Ulcerative Colitis-Derived Colonoid Culture: A Multi-Mineral-Approach to Improve Barrier Protein Expression |
| title_sort | ulcerative colitis derived colonoid culture a multi mineral approach to improve barrier protein expression |
| topic | basement membrane calcium cell barrier colonoid desmosome organoid culture |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2020.577221/full |
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