Metabolite profiling and pharmacokinetic evaluation of hydrocortisone in a perfused 3D human liver bioreactor
Endotoxin lipopolysaccharide (LPS) is known to cause liver injury primarily involving inflammatory cells such as Kupffer cells, but few in vitro culture models are applicable for investigation of inflammatory effects on drug metabolism. We have developed a 3D human microphysiological hepatocyte-Kupf...
| Main Authors: | , , , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
American Society for Pharmacology & Experimental Therapeutics (ASPET)
2015
|
| Online Access: | http://hdl.handle.net/1721.1/96927 https://orcid.org/0000-0002-1801-5548 https://orcid.org/0000-0002-2325-552X https://orcid.org/0000-0002-9618-8495 |
| _version_ | 1826206453310947328 |
|---|---|
| author | Sarkar, Ujjal Rivera-Burgos, Dinelia Large, Emma M. Hughes, David J. Kodihalli, Ravindra Dyer, Rachel Lee Ebrahimkhani, Mohammad Reza Wishnok, John S. Griffith, Linda G. Tannenbaum, Steven Robert |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Sarkar, Ujjal Rivera-Burgos, Dinelia Large, Emma M. Hughes, David J. Kodihalli, Ravindra Dyer, Rachel Lee Ebrahimkhani, Mohammad Reza Wishnok, John S. Griffith, Linda G. Tannenbaum, Steven Robert |
| author_sort | Sarkar, Ujjal |
| collection | MIT |
| description | Endotoxin lipopolysaccharide (LPS) is known to cause liver injury primarily involving inflammatory cells such as Kupffer cells, but few in vitro culture models are applicable for investigation of inflammatory effects on drug metabolism. We have developed a 3D human microphysiological hepatocyte-Kupffer-cell coculture system and evaluated the anti-inflammatory effect of glucocorticoids on liver cultures. LPS was introduced to the cultures to elicit an inflammatory response and assessed by the release of pro-inflammatory cytokines, IL6 and TNFα. A sensitive and specific RP-UHPLC-QTOF-MS method was used to evaluate hydrocortisone disappearance and metabolism at near physiological levels. For this, the systems were dosed with 100 nM hydrocortisone and circulated for two days; hydrocortisone was depleted to approximately 30 nM, with first-order kinetics. Phase I metabolites, including tetrahydrocortisone and dihydrocortisol, accounted for 8-10 % of the loss, and 45-52 % was phase II metabolites, including glucuronides of tetrahydrocortisol and tetrahydrocortisone. Pharmacokinetic parameters, i.e., half-life (t1/2), rate of elimination (kel), clearance (CL), and area under the curve (AUC), were 23.03 h, 0.03 h-1, 6.6x10-5 L. h-1 and 1.03 mg/L*h respectively. The ability of the bioreactor to predict the in vivo clearance of hydrocortisone was characterized and the obtained intrinsic clearance values correlated with human data. This system offers a physiologically-relevant tool for investigating hepatic function in an inflamed liver. Endotoxin lipopolysaccharide (LPS) is known to cause liver injury primarily involving inflammatory cells such as Kupffer cells, but few in vitro culture models are applicable for investigation of inflammatory effects on drug metabolism. We have developed a 3D human microphysiological hepatocyte-Kupffer-cell coculture system and evaluated the anti-inflammatory effect of glucocorticoids on liver cultures. LPS was introduced to the cultures to elicit an inflammatory response and assessed by the release of pro-inflammatory cytokines, IL6 and TNFα. A sensitive and specific RP-UHPLC-QTOF-MS method was used to evaluate hydrocortisone disappearance and metabolism at near physiological levels. For this, the systems were dosed with 100 nM hydrocortisone and circulated for two days; hydrocortisone was depleted to approximately 30 nM, with first-order kinetics. Phase I metabolites, including tetrahydrocortisone and dihydrocortisol, accounted for 8-10 % of the loss, and 45-52 % was phase II metabolites, including glucuronides of tetrahydrocortisol and tetrahydrocortisone. Pharmacokinetic parameters, i.e., half-life (t[subscript 1/2]), rate of elimination (k[subscript el]), clearance (CL), and area under the curve (AUC), were 23.03 h, 0.03 h[superscript -1], 6.6x10[superscript -5] L. h-1 and 1.03 mg/L*h respectively. The ability of the bioreactor to predict the in vivo clearance of hydrocortisone was characterized and the obtained intrinsic clearance values correlated with human data. This system offers a physiologically-relevant tool for investigating hepatic function in an inflamed liver. |
| first_indexed | 2024-09-23T13:30:55Z |
| format | Article |
| id | mit-1721.1/96927 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:30:55Z |
| publishDate | 2015 |
| publisher | American Society for Pharmacology & Experimental Therapeutics (ASPET) |
| record_format | dspace |
| spelling | mit-1721.1/969272022-10-01T15:35:46Z Metabolite profiling and pharmacokinetic evaluation of hydrocortisone in a perfused 3D human liver bioreactor Sarkar, Ujjal Rivera-Burgos, Dinelia Large, Emma M. Hughes, David J. Kodihalli, Ravindra Dyer, Rachel Lee Ebrahimkhani, Mohammad Reza Wishnok, John S. Griffith, Linda G. Tannenbaum, Steven Robert Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Chemistry Massachusetts Institute of Technology. Department of Mechanical Engineering Sarkar, Ujjal Rivera-Burgos, Dinelia Kodihalli, Ravindra Dyer, Rachel Lee Ebrahimkhani, Mohammad Reza Wishnok, John S. Griffith, Linda G. Tannenbaum, Steven Robert Endotoxin lipopolysaccharide (LPS) is known to cause liver injury primarily involving inflammatory cells such as Kupffer cells, but few in vitro culture models are applicable for investigation of inflammatory effects on drug metabolism. We have developed a 3D human microphysiological hepatocyte-Kupffer-cell coculture system and evaluated the anti-inflammatory effect of glucocorticoids on liver cultures. LPS was introduced to the cultures to elicit an inflammatory response and assessed by the release of pro-inflammatory cytokines, IL6 and TNFα. A sensitive and specific RP-UHPLC-QTOF-MS method was used to evaluate hydrocortisone disappearance and metabolism at near physiological levels. For this, the systems were dosed with 100 nM hydrocortisone and circulated for two days; hydrocortisone was depleted to approximately 30 nM, with first-order kinetics. Phase I metabolites, including tetrahydrocortisone and dihydrocortisol, accounted for 8-10 % of the loss, and 45-52 % was phase II metabolites, including glucuronides of tetrahydrocortisol and tetrahydrocortisone. Pharmacokinetic parameters, i.e., half-life (t1/2), rate of elimination (kel), clearance (CL), and area under the curve (AUC), were 23.03 h, 0.03 h-1, 6.6x10-5 L. h-1 and 1.03 mg/L*h respectively. The ability of the bioreactor to predict the in vivo clearance of hydrocortisone was characterized and the obtained intrinsic clearance values correlated with human data. This system offers a physiologically-relevant tool for investigating hepatic function in an inflamed liver. Endotoxin lipopolysaccharide (LPS) is known to cause liver injury primarily involving inflammatory cells such as Kupffer cells, but few in vitro culture models are applicable for investigation of inflammatory effects on drug metabolism. We have developed a 3D human microphysiological hepatocyte-Kupffer-cell coculture system and evaluated the anti-inflammatory effect of glucocorticoids on liver cultures. LPS was introduced to the cultures to elicit an inflammatory response and assessed by the release of pro-inflammatory cytokines, IL6 and TNFα. A sensitive and specific RP-UHPLC-QTOF-MS method was used to evaluate hydrocortisone disappearance and metabolism at near physiological levels. For this, the systems were dosed with 100 nM hydrocortisone and circulated for two days; hydrocortisone was depleted to approximately 30 nM, with first-order kinetics. Phase I metabolites, including tetrahydrocortisone and dihydrocortisol, accounted for 8-10 % of the loss, and 45-52 % was phase II metabolites, including glucuronides of tetrahydrocortisol and tetrahydrocortisone. Pharmacokinetic parameters, i.e., half-life (t[subscript 1/2]), rate of elimination (k[subscript el]), clearance (CL), and area under the curve (AUC), were 23.03 h, 0.03 h[superscript -1], 6.6x10[superscript -5] L. h-1 and 1.03 mg/L*h respectively. The ability of the bioreactor to predict the in vivo clearance of hydrocortisone was characterized and the obtained intrinsic clearance values correlated with human data. This system offers a physiologically-relevant tool for investigating hepatic function in an inflamed liver. United States. Defense Advanced Research Projects Agency (DARPA-BAA-11-73 Microphysiological Systems W911NF-12-2-0039) National Institutes of Health (U.S.) (5-UH2-TR000496) Massachusetts Institute of Technology. Center for Environmental Health Sciences (P30-ES002109) 2015-05-07T14:40:00Z 2015-05-07T14:40:00Z 2015-04 Article http://purl.org/eprint/type/JournalArticle 1521-009X 0090-9556 http://hdl.handle.net/1721.1/96927 Sarkar, U., D. Rivera-Burgos, E. M. Large, D. J. Hughes, K. C. Ravindra, R. L. Dyer, M. R. Ebrahimkhani, J. S. Wishnok, L. G. Griffith, and S. R. Tannenbaum. “Metabolite Profiling and Pharmacokinetic Evaluation of Hydrocortisone in a Perfused 3D Human Liver Bioreactor.” Drug Metabolism and Disposition (April 29, 2015). https://orcid.org/0000-0002-1801-5548 https://orcid.org/0000-0002-2325-552X https://orcid.org/0000-0002-9618-8495 en_US http://dx.doi.org/10.1124/dmd.115.063495 Drug Metabolism and Disposition Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf American Society for Pharmacology & Experimental Therapeutics (ASPET) Tannenbaum |
| spellingShingle | Sarkar, Ujjal Rivera-Burgos, Dinelia Large, Emma M. Hughes, David J. Kodihalli, Ravindra Dyer, Rachel Lee Ebrahimkhani, Mohammad Reza Wishnok, John S. Griffith, Linda G. Tannenbaum, Steven Robert Metabolite profiling and pharmacokinetic evaluation of hydrocortisone in a perfused 3D human liver bioreactor |
| title | Metabolite profiling and pharmacokinetic evaluation of hydrocortisone in a perfused 3D human liver bioreactor |
| title_full | Metabolite profiling and pharmacokinetic evaluation of hydrocortisone in a perfused 3D human liver bioreactor |
| title_fullStr | Metabolite profiling and pharmacokinetic evaluation of hydrocortisone in a perfused 3D human liver bioreactor |
| title_full_unstemmed | Metabolite profiling and pharmacokinetic evaluation of hydrocortisone in a perfused 3D human liver bioreactor |
| title_short | Metabolite profiling and pharmacokinetic evaluation of hydrocortisone in a perfused 3D human liver bioreactor |
| title_sort | metabolite profiling and pharmacokinetic evaluation of hydrocortisone in a perfused 3d human liver bioreactor |
| url | http://hdl.handle.net/1721.1/96927 https://orcid.org/0000-0002-1801-5548 https://orcid.org/0000-0002-2325-552X https://orcid.org/0000-0002-9618-8495 |
| work_keys_str_mv | AT sarkarujjal metaboliteprofilingandpharmacokineticevaluationofhydrocortisoneinaperfused3dhumanliverbioreactor AT riveraburgosdinelia metaboliteprofilingandpharmacokineticevaluationofhydrocortisoneinaperfused3dhumanliverbioreactor AT largeemmam metaboliteprofilingandpharmacokineticevaluationofhydrocortisoneinaperfused3dhumanliverbioreactor AT hughesdavidj metaboliteprofilingandpharmacokineticevaluationofhydrocortisoneinaperfused3dhumanliverbioreactor AT kodihalliravindra metaboliteprofilingandpharmacokineticevaluationofhydrocortisoneinaperfused3dhumanliverbioreactor AT dyerrachellee metaboliteprofilingandpharmacokineticevaluationofhydrocortisoneinaperfused3dhumanliverbioreactor AT ebrahimkhanimohammadreza metaboliteprofilingandpharmacokineticevaluationofhydrocortisoneinaperfused3dhumanliverbioreactor AT wishnokjohns metaboliteprofilingandpharmacokineticevaluationofhydrocortisoneinaperfused3dhumanliverbioreactor AT griffithlindag metaboliteprofilingandpharmacokineticevaluationofhydrocortisoneinaperfused3dhumanliverbioreactor AT tannenbaumstevenrobert metaboliteprofilingandpharmacokineticevaluationofhydrocortisoneinaperfused3dhumanliverbioreactor |