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Article: Dysregulation of the mRNA Expression of Human Renal Drug Transporters by Proinflammatory Cytokines in Primary Human Proximal Tubular Epithelial Cells.

Tsang, Yik Pui / Hao, Tianran / Mao, Qingcheng / Kelly, Edward J / Unadkat, Jashvant D

2024 Volume 16, Issue 2

Abstract: Proinflammatory cytokines, which are elevated during inflammation or infections, can affect drug pharmacokinetics (PK) due to the altered expression or activity of drug transporters and/or metabolizing enzymes. To date, such studies have focused on the ... ...

Abstract	Proinflammatory cytokines, which are elevated during inflammation or infections, can affect drug pharmacokinetics (PK) due to the altered expression or activity of drug transporters and/or metabolizing enzymes. To date, such studies have focused on the effect of cytokines on the activity and/or mRNA expression of hepatic transporters and drug-metabolizing enzymes. However, many antibiotics and antivirals used to treat infections are cleared by renal transporters, including the basal organic cation transporter 2 (OCT2), organic anion transporters 1 and 3 (OAT1 and 3), the apical multidrug and toxin extrusion proteins 1 and 2-K (MATE1/2-K), and multidrug resistance-associated protein 2 and 4 (MRP2/4). Here, we determined the concentration-dependent effect of interleukin-6 (IL-6), IL-1β, tumor necrosis factor (TNF)-α, and interferon-γ (IFN-γ) on the mRNA expression of human renal transporters in freshly isolated primary human renal proximal tubular epithelial cells (PTECs,
Language	English
Publishing date	2024-02-16
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527217-2
ISSN	1999-4923
ISSN	1999-4923
DOI	10.3390/pharmaceutics16020285
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Article ; Online: Endothelial dysfunction is dampened by early administration of fresh frozen plasma in a rodent burn shock model.

Kelly, Edward J / Ziedins, Eriks E / Carney, Bonnie C / Moffatt, Lauren T / Shupp, Jeffrey W

The journal of trauma and acute care surgery

2024

Abstract: Background: Endothelial dysfunction has been implicated in the pathogenesis of burn shock affecting patients with large thermal injury. In response to injury, glycocalyx components like Syndecan-1 (SDC-1) are shed into circulation and have been used as ... ...

Abstract	Background: Endothelial dysfunction has been implicated in the pathogenesis of burn shock affecting patients with large thermal injury. In response to injury, glycocalyx components like Syndecan-1 (SDC-1) are shed into circulation and have been used as markers of endothelial damage. Previous work in our lab has shown plasma inclusive resuscitation (PIR) with fresh frozen plasma (FFP) ameliorates endothelial damage. However, there remains a paucity of information regarding optimal timing and dosing of PIR as well as organ specific endothelial responses to shock. We aimed to examine the impact of PIR on endothelial dysfunction using clinically-translatable timing and dosing. Methods: Spraque-Dawley rats were used to create thermal burns. Rats were subjected to 40% TBSA scald burns and were resuscitated with LR only, LR + albumin and LR + early 1 ml boluses of FFP at 0,2,4 and 8 hours post-injury. A late group also received LR + FFP starting at hour 10 post-injury. SDC-1 levels were quantified by ELISA and qRT-PCR analysis characterized transcription of glycocalyx components and inflammatory cytokines in the lung and spleen. Evan's blue dye (EBD) was used to quantify amount of vascular leakage. Results: LR + early FFP reduced EBD extravasation when compared to LR only groups while late FFP did not. When comparing LR only vs LR + early FFP, SDC-1 levels were reduced in the LR + early FFP group at hour 8, 12 and 24 (5.23 vs. 2.07, p < 0.001, 4.49 vs. 2.05, p < 0.01, 3.82 vs 2.08, p < 0.05, respectively). LR only groups had upregulation of Exostosin-1 (EXT-1) and SDC-1 in the lung compared to LR + early FFP groups (p < 0.01 and p < 0.05) as well as upregulation of cytokines IL-10 and IFN-Ƴ (p < 0.001 and p < 0.001). Conclusions: Early administration of LR + FFP reduces the magnitude of SDC-1 shedding and dampens the cytokine response to injury. The upregulation of glycocalyx components as a response to endothelial injury is also decreased in the lung and spleen by LR + early FFP administration.
Language	English
Publishing date	2024-05-20
Publishing country	United States
Document type	Journal Article
ZDB-ID	2651070-4
ISSN	2163-0763 ; 2163-0755
ISSN (online)	2163-0763
ISSN	2163-0755
DOI	10.1097/TA.0000000000004373
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Zs.A 127: Show issues

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Article ; Online: Molecular Mechanisms of Organic Anion Transporting Polypeptide-Mediated Organic Anion Clearance at the Blood-Cerebrospinal Fluid Barrier.

Sun, Austin / Hagenbuch, Bruno / Kelly, Edward J / Wang, Joanne

Molecular pharmacology

2023 Volume 104, Issue 6, Page(s) 255–265

Abstract: The blood-cerebrospinal fluid barrier (BCSFB), formed by the choroid plexus epithelial (CPE) cells, plays an active role in removing drugs and metabolic wastes from the brain. Recent functional studies in isolated mouse choroid plexus (CP) tissues ... ...

Abstract	The blood-cerebrospinal fluid barrier (BCSFB), formed by the choroid plexus epithelial (CPE) cells, plays an active role in removing drugs and metabolic wastes from the brain. Recent functional studies in isolated mouse choroid plexus (CP) tissues suggested the presence of organic anion transporting polypeptides (OATPs, encoded by SLCOs) at the apical membrane of BCSFB, which may clear large organic anions from the cerebrospinal fluid (CSF). However, the specific OATP isoform involved is unclear. Using quantitative fluorescence imaging, we showed that the fluorescent anions sulforhodamine 101 (SR101), fluorescein methotrexate (FL-MTX), and 8-fluorescein-cAMP (fluo-cAMP) are actively transported from the CSF to the subepithelial space in CP tissues isolated from wild-type mice. In contrast, transepithelial transport of these compounds across the CPE cells was abolished in Oatp1a/1b
MeSH term(s)	Mice ; Humans ; Animals ; Kinetics ; Blood-Brain Barrier/metabolism ; Organic Anion Transporters/metabolism ; Biological Transport ; Fluorescein/metabolism ; Anions/metabolism
Chemical Substances	fluorescein-methotrexate ; Organic Anion Transporters ; Fluorescein (TPY09G7XIR) ; Anions
Language	English
Publishing date	2023-08-31
Publishing country	United States
Document type	Journal Article
ZDB-ID	124034-1
ISSN	1521-0111 ; 0026-895X
ISSN (online)	1521-0111
ISSN	0026-895X
DOI	10.1124/molpharm.123.000703
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Zs.A 525: Show issues

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Article: Kidney microphysiological models for nephrotoxicity assessment.

Mahadeo, Anish / Yeung, Catherine K / Himmelfarb, Jonathan / Kelly, Edward J

Current opinion in toxicology

2022 Volume 30

Abstract: Nephrotoxicity testing is an important step in preclinical development of new molecular entities (NMEs) and has traditionally been performed in 2-D cell culture systems and animal models. However, 2-D culture systems fail to replicate ... ...

Abstract	Nephrotoxicity testing is an important step in preclinical development of new molecular entities (NMEs) and has traditionally been performed in 2-D cell culture systems and animal models. However, 2-D culture systems fail to replicate complex
Language	English
Publishing date	2022-03-30
Publishing country	Netherlands
Document type	Journal Article
ISSN	2468-2934
ISSN	2468-2934
DOI	10.1016/j.cotox.2022.03.002
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Article ; Online: Human Enteroid Monolayers: A Novel, Functionally-Stable Model for Investigating Oral Drug Disposition.

Arian, Christopher / Mahony, Eimear O / MacDonald, James W / Bammler, Theo K / Donowitz, Mark / Kelly, Edward J / Thummel, Kenneth E

Drug metabolism and disposition: the biological fate of chemicals

2024

Abstract: To further the development of an in vitro model which faithfully recapitulates drug disposition of orally administered drugs, we investigated the utility of human enteroid monolayers to simultaneously assess intestinal drug absorption and first-pass ... ...

Abstract	To further the development of an in vitro model which faithfully recapitulates drug disposition of orally administered drugs, we investigated the utility of human enteroid monolayers to simultaneously assess intestinal drug absorption and first-pass metabolism processes. We cultured human enteroid monolayers from three donors, derived via biopsies containing duodenal stem cells that were propagated and then differentiated atop permeable Transwell® inserts, and confirmed transformation into a largely enterocyte population via RNA-seq analysis and immunocytochemical (ICC) assays. Proper cell morphology was assessed and confirmed via bright field microscopy and ICC imaging of tight junction proteins and other apically and basolaterally localized proteins. Enteroid monolayer barrier integrity was demonstrated by elevated transepithelial electrical resistance (TEER) that stabilized after 10 days in culture and persisted for 42 days. These results were corroborated by low paracellular transport probe permeability at 7 and 21 days in culture. The activity of a prominent drug metabolizing enzyme, CYP3A, was confirmed at 7, 21, and 42 days culture under basal, 1α,25(OH)2 vitamin D3-induced, and 6',7'-dihydroxybergamottin-inhibited conditions. The duration of these experiments is particularly noteworthy, as this is the first study assessing drug metabolizing enzymes and transporters (DMET) expression/function for enteroids cultured for greater than 12 days. The sum of these results suggests enteroid monolayers are a promising ex vivo model to investigate and quantitatively predict an orally administered drug's intestinal absorption and/or metabolism.
Language	English
Publishing date	2024-05-14
Publishing country	United States
Document type	Journal Article
ZDB-ID	186795-7
ISSN	1521-009X ; 0090-9556
ISSN (online)	1521-009X
ISSN	0090-9556
DOI	10.1124/dmd.124.001551
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Zs.A 1014: Show issues

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Article ; Online: Endothelial damage occurs early after inhalation injury as measured by increased syndecan-1 levels.

Kelly, Edward J / Carney, Bonnie C / Oliver, Mary A / Keyloun, John W / Prindeze, Nicholas J / Nisar, Saira / Moffatt, Lauren T / Shupp, Jeffrey W

Journal of burn care & research : official publication of the American Burn Association

2023 Volume 44, Issue 4, Page(s) 769–774

Abstract: Inhalation injury is a significant cause of morbidity and mortality in the burn patient population. However, the pathogenesis of inhalation injury and its potential involvement in burn shock is not well understood. Preclinical studies have shown ... ...

Abstract	Inhalation injury is a significant cause of morbidity and mortality in the burn patient population. However, the pathogenesis of inhalation injury and its potential involvement in burn shock is not well understood. Preclinical studies have shown endothelial injury, as measured by syndecan-1 (SDC-1) levels, to be involved in the increased vascular permeability seen in shock states. Furthermore, the lung has been identified as a site of significant SDC-1 shedding. Here we aim to characterize the contribution of endotheliopathy caused by inhalation alone in a swine model. When comparing injured animals, the fold change of circulating SDC-1 levels from preinjury was significantly higher at 2, 4, and 6 hours postinjury (P = .0045, P = .0017, and P < .001, respectively). When comparing control animals, the fold change of SDC-1 from preinjury was not significant at any timepoint. When comparing injured animals versus controls, the fold change of SDC-1 injured animals was significantly greater at 2, 4, 6, and 18 hours (P = .004, P = .03, P < .001, and P = .03, respectively). Histological sections showed higher lung injury severity compared to control uninjured lungs (0.56 vs 0.38, P < .001). This novel animal model shows significant increases in SDC-1 levels that provide evidence for the connection between smoke inhalation injury and endothelial injury. Further understanding of the mechanisms underlying inhalation injury and its contribution to shock physiology may aid in development of early, more targeted therapies.
MeSH term(s)	Humans ; Animals ; Swine ; Burns/therapy ; Lung Injury/etiology ; Lung Injury/pathology ; Syndecan-1 ; Lung/pathology ; Smoke Inhalation Injury/pathology
Chemical Substances	Syndecan-1
Language	English
Publishing date	2023-03-16
Publishing country	England
Document type	Journal Article
ZDB-ID	2224246-6
ISSN	1559-0488 ; 1559-047X
ISSN (online)	1559-0488
ISSN	1559-047X
DOI	10.1093/jbcr/irad018
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Zs.A 6223: Show issues

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Article ; Online: Neutrophil Extracellular Traps Are Induced by Coronavirus 2019 Disease-Positive Patient Plasma and Persist Longitudinally: A Possible Link to Endothelial Dysfunction as Measured by Syndecan-1.

Kelly, Edward J / Oliver, Mary A / Carney, Bonnie C / Kolachana, Sindhura / Moffatt, Lauren T / Shupp, Jeffrey W

Surgical infections

2023 Volume 24, Issue 10, Page(s) 887–896

Abstract: Background: ...

Abstract	Background:
MeSH term(s)	Humans ; COVID-19 ; Extracellular Traps/chemistry ; Extracellular Traps/metabolism ; Histones ; Neutrophils ; Peroxidase/analysis ; Peroxidase/metabolism ; Syndecan-1
Chemical Substances	Histones ; Peroxidase (EC 1.11.1.7) ; Syndecan-1 ; SDC1 protein, human
Language	English
Publishing date	2023-11-21
Publishing country	United States
Document type	Journal Article
ZDB-ID	1440120-4
ISSN	1557-8674 ; 1096-2964
ISSN (online)	1557-8674
ISSN	1096-2964
DOI	10.1089/sur.2023.156
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Zs.A 5617: Show issues

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Article ; Online: Examining Obesity and Its Association With Burn Injury: A Secondary Analysis of the Transfusion Requirement in Burn Care Evaluation Study.

Kelly, Edward J / Reese, Adam D / Carney, Bonnie C / Keyloun, John W / Palmieri, Tina L / Moffatt, Lauren T / Shupp, Jeffrey W / Tejiram, Shawn

The Journal of surgical research

2023 Volume 290, Page(s) 221–231

Abstract: Introduction: Literature examining the connection between obesity and burn injuries is limited. This study is a secondary analysis of a multicenter trial data set to investigate the association between burn outcomes and obesity following severe burn ... ...

Abstract	Introduction: Literature examining the connection between obesity and burn injuries is limited. This study is a secondary analysis of a multicenter trial data set to investigate the association between burn outcomes and obesity following severe burn injury. Materials and methods: Body mass index (BMI) was used to stratify patients as normal weight (NW; BMI 18.5-25), all obese (AO; any BMI>30), obese I (OI; BMI 30-34.9), obese II (OII; BMI 35-39.9), or obese III (OIII; BMI>40). The primary outcome examined was mortality. Secondary outcomes included hospital length of stay (LOS), number of transfusions, injury scores, infection occurrences, number of operations, ventilator days, intensive care unit LOS, and days to wound healing. Results: Of 335 patients included for study, 130 were obese. Median total body surface area (TBSA) was 31%, 77 patients (23%) had inhalation injury and 41 patients died. Inhalation injury was higher in OIII than NW (42.1% versus 20%, P = 0.03). Blood stream infections (BSI) were higher in OI versus NW (0.72 versus 0.33, P = 0.03). Total operations, ventilator days, days to wound healing, multiorgan dysfunction score, Acute Physiology and Chronic Health Evaluationscore, hospital LOS, and intensive care unit LOS were not significantly affected by BMI classification. Mortality was not significantly different between obesity groups. Kaplan-Meier survival curves did not significantly differ between the groups (χ Conclusions: No significant association between obesity and mortality was seen after burn injury. Age, TBSA, and percent full- thickness burn were independent predictors of mortality after burn injury, while BMI classification was not.
MeSH term(s)	Humans ; Burns/complications ; Burns/therapy ; Obesity/complications ; Obesity/epidemiology ; Obesity/therapy ; Blood Transfusion ; Sepsis/complications ; Organ Dysfunction Scores ; Retrospective Studies ; Length of Stay
Language	English
Publishing date	2023-06-06
Publishing country	United States
Document type	Multicenter Study ; Journal Article
ZDB-ID	80170-7
ISSN	1095-8673 ; 0022-4804
ISSN (online)	1095-8673
ISSN	0022-4804
DOI	10.1016/j.jss.2023.05.005
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Zs.A 178: Show issues

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Article ; Online: Mechanisms of CYP3A Induction During Pregnancy: Studies in HepaRG Cells.

Sachar, Madhav / Kelly, Edward J / Unadkat, Jashvant D

The AAPS journal

2019 Volume 21, Issue 3, Page(s) 45

Abstract: Activity of CYP3A, an enzyme responsible for metabolism of many marketed drugs, is induced by ~ 2-fold in pregnant women. Through studies in sandwich-cultured human hepatocytes (SCHH) and HepaRG cells, our laboratory has shown that this induction is ... ...

Abstract	Activity of CYP3A, an enzyme responsible for metabolism of many marketed drugs, is induced by ~ 2-fold in pregnant women. Through studies in sandwich-cultured human hepatocytes (SCHH) and HepaRG cells, our laboratory has shown that this induction is likely mediated by the increase in cortisol plasma concentrations during pregnancy. Cortisol, at plasma concentrations observed during the third trimester (~ 800 nM), either alone or in combination with other pregnancy-related hormones, induces CYP3A activity in SCHH and HepaRG cells when cultured in dexamethasone-free media. To determine the mechanism(s) by which cortisol induces CYP3A activity, HepaRG cells were pre-incubated in dexamethasone-free medium and then incubated for 72 h with cortisol (798 nM). Glucocorticoid receptor (GR), pregnane X receptor (PXR), and CYP3A4 or CYP3A5 were knocked down using siRNA, and mRNA expression of these genes was measured. CYP3A4, and not CYP3A5, was found to be the dominant contributor to total CYP3A activity in control- and cortisol-treated HepaRG cells. Constitutive mRNA expression of CYP3A4 in HepaRG cells was regulated by both PXR and GR whereas constitutive expression of CYP3A5 in HepaRG cells was regulated by GR alone. Cortisol-mediated CYP3A4 induction in HepaRG cells was primarily mediated by GR-dependent PXR induction pathway and to a smaller extent via a PXR-independent pathway. Cortisol-mediated CYP3A5 induction was regulated by GR-dependent PXR-independent pathway. These data indicate that PXR plays a central role in cortisol-mediated induction of CYP3A activity during pregnancy and suggests that other enzymes and transporters, such as CYP2B6 and P-glycoprotein, may also be induced during pregnancy via the same mechanism(s).
MeSH term(s)	Cell Line ; Cytochrome P-450 CYP3A/genetics ; Cytochrome P-450 CYP3A/metabolism ; Female ; Gene Knockdown Techniques ; Humans ; Hydrocortisone/blood ; Hydrocortisone/metabolism ; Pregnancy/blood ; Pregnancy/metabolism ; Pregnane X Receptor/genetics ; Pregnane X Receptor/metabolism ; RNA, Small Interfering/metabolism ; Receptors, Glucocorticoid/genetics ; Receptors, Glucocorticoid/metabolism ; Up-Regulation
Chemical Substances	NR1I2 protein, human ; Pregnane X Receptor ; RNA, Small Interfering ; Receptors, Glucocorticoid ; CYP3A5 protein, human (EC 1.14.14.1) ; Cytochrome P-450 CYP3A (EC 1.14.14.1) ; CYP3A4 protein, human (EC 1.14.14.55) ; Hydrocortisone (WI4X0X7BPJ)
Language	English
Publishing date	2019-03-27
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1550-7416
ISSN (online)	1550-7416
DOI	10.1208/s12248-019-0316-z
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Article: Development of a kidney microphysiological system hardware platform for microgravity studies.

Jones-Isaac, Kendan A / Lidberg, Kevin A / Yeung, Catherine K / Yang, Jade / Bain, Jacelyn / Ruiz, Micaela / Koenig, Greta / Koenig, Paul / Countryman, Stefanie / Himmelfarb, Jonathan / Kelly, Edward J

NPJ microgravity

2024 Volume 10, Issue 1, Page(s) 54

Abstract: Determining the physiological effects of microgravity on the human kidney is limited to relatively insensitive tests of biofluids (blood and urine) that do not return abnormal results until more than 50% of kidney function is lost. We have developed an " ... ...

Abstract	Determining the physiological effects of microgravity on the human kidney is limited to relatively insensitive tests of biofluids (blood and urine) that do not return abnormal results until more than 50% of kidney function is lost. We have developed an "organ on chip" microphysiological model of the human kidney proximal tubule (PT-MPS) that can recapitulate many kidney functions and disease states and could play a critical role in determining mechanisms of early kidney dysfunction in microgravity. However, the ground-based PT-MPS system is incompatible with spaceflight as it requires a large pneumatic system coupled to a cell incubator for perfusion and intensive hand-on manipulation. Herein, we report the hardware engineering and performance of the Kidney Chip Perfusion Platform (KCPP), a small, advanced, semi-autonomous hardware platform to support kidney microphysiological model experiments in microgravity. The KCPP is composed of five components, the kidney MPS, the MPS housing and valve block, media cassettes, fixative cassettes, and the programable precision syringe pump. The system has been deployed twice to the ISSNL (aboard CRS-17 and CRS-22). From each set of ISSNL experiments and ground-based controls, we were able to recover PT-MPS effluent for biomarker analysis and RNA suitable for transcriptomics analysis demonstrating the usability and functionality of the KCPP.
Language	English
Publishing date	2024-05-11
Publishing country	United States
Document type	Journal Article
ZDB-ID	2823626-9
ISSN	2373-8065
ISSN	2373-8065
DOI	10.1038/s41526-024-00398-0
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