Hypoxia Primes Human ISCs for Interleukin-Dependent Rescue of Stem Cell ActivitySummary
Background and Aims: Hypoxia in the intestinal epithelium can be caused by acute ischemic events or chronic inflammation in which immune cell infiltration produces inflammatory hypoxia starving the mucosa of oxygen. The epithelium has the capacity to regenerate after some ischemic and inflammatory c...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-01-01
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| Series: | Cellular and Molecular Gastroenterology and Hepatology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352345X23001443 |
| _version_ | 1797676613077303296 |
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| author | Kristina R. Rivera R. Jarrett Bliton Joseph Burclaff Michael J. Czerwinski Jintong Liu Jessica M. Trueblood Caroline M. Hinesley Keith A. Breau Halston E. Deal Shlok Joshi Vladimir A. Pozdin Ming Yao Amanda L. Ziegler Anthony T. Blikslager Michael A. Daniele Scott T. Magness |
| author_facet | Kristina R. Rivera R. Jarrett Bliton Joseph Burclaff Michael J. Czerwinski Jintong Liu Jessica M. Trueblood Caroline M. Hinesley Keith A. Breau Halston E. Deal Shlok Joshi Vladimir A. Pozdin Ming Yao Amanda L. Ziegler Anthony T. Blikslager Michael A. Daniele Scott T. Magness |
| author_sort | Kristina R. Rivera |
| collection | DOAJ |
| description | Background and Aims: Hypoxia in the intestinal epithelium can be caused by acute ischemic events or chronic inflammation in which immune cell infiltration produces inflammatory hypoxia starving the mucosa of oxygen. The epithelium has the capacity to regenerate after some ischemic and inflammatory conditions suggesting that intestinal stem cells (ISCs) are highly tolerant to acute and chronic hypoxia; however, the impact of hypoxia on human ISC (hISC) function has not been reported. Here we present a new microphysiological system (MPS) to investigate how hypoxia affects hISCs from healthy donors and test the hypothesis that prolonged hypoxia modulates how hISCs respond to inflammation-associated interleukins (ILs). Methods: hISCs were exposed to <1.0% oxygen in the MPS for 6, 24, 48, and 72 hours. Viability, hypoxia-inducible factor 1a (HIF1a) response, transcriptomics, cell cycle dynamics, and response to cytokines were evaluated in hISCs under hypoxia. HIF stabilizers and inhibitors were screened to evaluate HIF-dependent responses. Results: The MPS enables precise, real-time control and monitoring of oxygen levels at the cell surface. Under hypoxia, hISCs maintain viability until 72 hours and exhibit peak HIF1a at 24 hours. hISC activity was reduced at 24 hours but recovered at 48 hours. Hypoxia induced increases in the proportion of hISCs in G1 and expression changes in 16 IL receptors. Prolyl hydroxylase inhibition failed to reproduce hypoxia-dependent IL-receptor expression patterns. hISC activity increased when treated IL1β, IL2, IL4, IL6, IL10, IL13, and IL25 and rescued hISC activity caused by 24 hours of hypoxia. Conclusions: Hypoxia pushes hISCs into a dormant but reversible proliferative state and primes hISCs to respond to a subset of ILs that preserves hISC activity. These findings have important implications for understanding intestinal epithelial regeneration mechanisms caused by inflammatory hypoxia. |
| first_indexed | 2024-03-11T22:31:45Z |
| format | Article |
| id | doaj.art-4c003ca116b84e8fb51d880c837e2004 |
| institution | Directory Open Access Journal |
| issn | 2352-345X |
| language | English |
| last_indexed | 2024-03-11T22:31:45Z |
| publishDate | 2023-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cellular and Molecular Gastroenterology and Hepatology |
| spelling | doaj.art-4c003ca116b84e8fb51d880c837e20042023-09-23T05:11:16ZengElsevierCellular and Molecular Gastroenterology and Hepatology2352-345X2023-01-01165823846Hypoxia Primes Human ISCs for Interleukin-Dependent Rescue of Stem Cell ActivitySummaryKristina R. Rivera0R. Jarrett Bliton1Joseph Burclaff2Michael J. Czerwinski3Jintong Liu4Jessica M. Trueblood5Caroline M. Hinesley6Keith A. Breau7Halston E. Deal8Shlok Joshi9Vladimir A. Pozdin10Ming Yao11Amanda L. Ziegler12Anthony T. Blikslager13Michael A. Daniele14Scott T. Magness15Joint Department of Biomedical Engineering, North Carolina State University and University of North Carolina at Chapel Hill, Raleigh, North CarolinaJoint Department of Biomedical Engineering, North Carolina State University and University of North Carolina at Chapel Hill, Raleigh, North CarolinaJoint Department of Biomedical Engineering, North Carolina State University and University of North Carolina at Chapel Hill, Raleigh, North CarolinaDepartment of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North CarolinaDepartment of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North CarolinaCenter for Gastrointestinal Biology and Disease, University of North Carolina at Chapel Hill, Chapel Hill, North CarolinaCenter for Gastrointestinal Biology and Disease, University of North Carolina at Chapel Hill, Chapel Hill, North CarolinaDepartment of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North CarolinaJoint Department of Biomedical Engineering, North Carolina State University and University of North Carolina at Chapel Hill, Raleigh, North CarolinaDepartment of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North CarolinaDepartment of Electrical and Computer Engineering, North Carolina State University, Raleigh, North CarolinaDepartment of Mechanical and Aerospace Engineering, North Carolina State University, Raleigh, North CarolinaComparative Medicine Institute, Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North CarolinaComparative Medicine Institute, Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North CarolinaJoint Department of Biomedical Engineering, North Carolina State University and University of North Carolina at Chapel Hill, Raleigh, North Carolina; Department of Electrical and Computer Engineering, North Carolina State University, Raleigh, North CarolinaJoint Department of Biomedical Engineering, North Carolina State University and University of North Carolina at Chapel Hill, Raleigh, North Carolina; Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina; Center for Gastrointestinal Biology and Disease, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina; Department of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina; Correspondence Address correspondence to: Scott T. Magness, PhD, University of North Carolina at Chapel Hill, 111 Mason Farm Rd. Rm. 4337 MBRB, CB#7032, Chapel Hill, North Carolina, 27599.Background and Aims: Hypoxia in the intestinal epithelium can be caused by acute ischemic events or chronic inflammation in which immune cell infiltration produces inflammatory hypoxia starving the mucosa of oxygen. The epithelium has the capacity to regenerate after some ischemic and inflammatory conditions suggesting that intestinal stem cells (ISCs) are highly tolerant to acute and chronic hypoxia; however, the impact of hypoxia on human ISC (hISC) function has not been reported. Here we present a new microphysiological system (MPS) to investigate how hypoxia affects hISCs from healthy donors and test the hypothesis that prolonged hypoxia modulates how hISCs respond to inflammation-associated interleukins (ILs). Methods: hISCs were exposed to <1.0% oxygen in the MPS for 6, 24, 48, and 72 hours. Viability, hypoxia-inducible factor 1a (HIF1a) response, transcriptomics, cell cycle dynamics, and response to cytokines were evaluated in hISCs under hypoxia. HIF stabilizers and inhibitors were screened to evaluate HIF-dependent responses. Results: The MPS enables precise, real-time control and monitoring of oxygen levels at the cell surface. Under hypoxia, hISCs maintain viability until 72 hours and exhibit peak HIF1a at 24 hours. hISC activity was reduced at 24 hours but recovered at 48 hours. Hypoxia induced increases in the proportion of hISCs in G1 and expression changes in 16 IL receptors. Prolyl hydroxylase inhibition failed to reproduce hypoxia-dependent IL-receptor expression patterns. hISC activity increased when treated IL1β, IL2, IL4, IL6, IL10, IL13, and IL25 and rescued hISC activity caused by 24 hours of hypoxia. Conclusions: Hypoxia pushes hISCs into a dormant but reversible proliferative state and primes hISCs to respond to a subset of ILs that preserves hISC activity. These findings have important implications for understanding intestinal epithelial regeneration mechanisms caused by inflammatory hypoxia.http://www.sciencedirect.com/science/article/pii/S2352345X23001443Inflammatory HypoxiaMicrophysiological SystemIntestinal Stem CellsStem Cell PrimingOxygen SensorCytokines |
| spellingShingle | Kristina R. Rivera R. Jarrett Bliton Joseph Burclaff Michael J. Czerwinski Jintong Liu Jessica M. Trueblood Caroline M. Hinesley Keith A. Breau Halston E. Deal Shlok Joshi Vladimir A. Pozdin Ming Yao Amanda L. Ziegler Anthony T. Blikslager Michael A. Daniele Scott T. Magness Hypoxia Primes Human ISCs for Interleukin-Dependent Rescue of Stem Cell ActivitySummary Cellular and Molecular Gastroenterology and Hepatology Inflammatory Hypoxia Microphysiological System Intestinal Stem Cells Stem Cell Priming Oxygen Sensor Cytokines |
| title | Hypoxia Primes Human ISCs for Interleukin-Dependent Rescue of Stem Cell ActivitySummary |
| title_full | Hypoxia Primes Human ISCs for Interleukin-Dependent Rescue of Stem Cell ActivitySummary |
| title_fullStr | Hypoxia Primes Human ISCs for Interleukin-Dependent Rescue of Stem Cell ActivitySummary |
| title_full_unstemmed | Hypoxia Primes Human ISCs for Interleukin-Dependent Rescue of Stem Cell ActivitySummary |
| title_short | Hypoxia Primes Human ISCs for Interleukin-Dependent Rescue of Stem Cell ActivitySummary |
| title_sort | hypoxia primes human iscs for interleukin dependent rescue of stem cell activitysummary |
| topic | Inflammatory Hypoxia Microphysiological System Intestinal Stem Cells Stem Cell Priming Oxygen Sensor Cytokines |
| url | http://www.sciencedirect.com/science/article/pii/S2352345X23001443 |
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