My research group studies the short and long term effects of drugs of abuse, their mechanism of action, toxicity, metabolism, addiction potential, as well as new therapeutic approaches to treat drug addiction. We also develop new analytical methods for novel psychoactive substances on the drug market for implementation in the forensic toxicology laboratory. We employ animal models to study behavioral effects, in combination with analytical methods and molecular biology techniques for determination of drug concentrations and neurobiological and genetic changes. Our research group has a wide network of national and international collaborators. For more information, visit our website: "Experimental Drug Abuse Research".
Bogen, Inger Lise; Boix, Fernando; Andersen, Jannike Mørch; Steinsland, Synne; Nerem, Elisabeth & Mørland, Jørg Gustav
(2023).
Heroin metabolism in human blood and its impact for the design of an immunotherapeutic approach against heroin effects.
Basic & Clinical Pharmacology & Toxicology.
ISSN 1742-7835.
133(4),
s. 418–427.
doi: 10.1111/bcpt.13926.
Fulltekst i vitenarkiv
Bergh, Marianne Skov-Skov; Bogen, Inger Lise; Garibay, Nancy & Baumann, Michael H.
(2021).
Pharmacokinetics and pharmacodynamics of cyclopropylfentanyl in male rats.
Psychopharmacology.
ISSN 0033-3158.
238,
s. 3629–3641.
doi: 10.1007/s00213-021-05981-x.
Fulltekst i vitenarkiv
Andersen, Jannike Mørch; Bogen, Inger Lise; Karinen, Ritva Anneli; Brochmann, Gerd-Wenche; Mørland, Jørg Gustav & Vindenes, Vigdis
[Vis alle 7 forfattere av denne artikkelen](2021).
Does the preparation for intravenous administration affect the composition of heroin injections? A controlled laboratory study.
Addiction.
ISSN 0965-2140.
116(11),
s. 3104–3112.
doi: 10.1111/add.15492.
Fulltekst i vitenarkiv
Bergh, Marianne Skov-Skov; Bogen, Inger Lise; Nerem, Elisabeth; Wohlfarth, Ariane; Wilson, Steven Ray Haakon & Øiestad, Åse Marit Leere
(2021).
Discovering the major metabolites of the three novel fentanyl analogues 3-methylcrotonylfentanyl, furanylbenzylfentanyl, and 4-fluorocyclopropylbenzylfentanyl for forensic case work.
Forensic Toxicology.
ISSN 1860-8965.
39,
s. 167–178.
doi: 10.1007/s11419-020-00560-9.
Harrison, Sean; Siller, Richard; Tanaka, Yoshiaki; Xiang, Yangfei; Patterson, Benjamin & Kempf, Henning
[Vis alle 29 forfattere av denne artikkelen](2020).
Scalable production of tissue-like vascularised liver organoids from human PSCs.
bioRxiv.
ISSN 2692-8205.
doi: 10.1101/2020.12.02.406835.
Bergh, Marianne Skov-Skov; Øiestad, Åse Marit Leere; Baumann, Michael H & Bogen, Inger Lise
(2020).
Selectivity and sensitivity of urine fentanyl test strips to detect fentanyl analogues in illicit drugs.
International journal of drug policy.
ISSN 0955-3959.
90(103065).
doi: 10.1016/j.drugpo.2020.103065.
Kongstorp, Mette; Bogen, Inger Lise; Stiris, Tom & Andersen, Jannike Mørch
(2019).
High accumulation of methadone compared with buprenorphine in fetal rat brain after maternal exposure.
Journal of Pharmacology and Experimental Therapeutics.
ISSN 0022-3565.
371(1),
s. 130–137.
doi: 10.1124/jpet.119.259531.
Bergh, Marianne Skov-Skov; Bogen, Inger Lise; Garibay, Nancy & Baumann, Michael H
(2019).
Evidence for nonlinear accumulation of the ultrapotent fentanyl analog, carfentanil, after systemic administration to male rats.
Neuropharmacology.
ISSN 0028-3908.
158.
doi: 10.1016/j.neuropharm.2019.04.002.
Bergh, Marianne Skov-Skov; Bogen, Inger Lise; Wohlfarth, Ariane; Wilson, Steven Ray Haakon & Øiestad, Åse Marit Leere
(2019).
Distinguishing Between Cyclopropylfentanyl and Crotonylfentanyl by Methods Commonly Available in the Forensic Laboratory.
Therapeutic Drug Monitoring.
ISSN 0163-4356.
41(4),
s. 519–527.
doi: 10.1097/FTD.0000000000000617.
Fulltekst i vitenarkiv
Kvello, Anne Marte Sjursen; Andersen, Jannike Mørch; Boix, Fernando; Mørland, Jørg & Bogen, Inger Lise
(2019).
The role of 6-acetylmorphine in heroin-induced reward and locomotor sensitization in mice.
Addiction Biology.
ISSN 1355-6215.s. 1–10.
doi: 10.1111/adb.12727.
Fulltekst i vitenarkiv
Avvisati, Riccardo; Bogen, Inger Lise; Andersen, Jannike Mørch; Vindenes, Vigdis; Mørland, Jørg & Badiani, Aldo
[Vis alle 7 forfattere av denne artikkelen](2019).
The active heroin metabolite 6-acetylmorphine has robust reinforcing effects as assessed by self-administration in the rat.
Neuropharmacology.
ISSN 0028-3908.
150,
s. 192–199.
doi: 10.1016/j.neuropharm.2018.12.023.
Kvello, Anne Marte Sjursen; Andersen, Jannike Mørch; Øiestad, Elisabeth Leere; Steinsland, Synne; Aase, Audun & Mørland, Jørg
[Vis alle 7 forfattere av denne artikkelen](2019).
A monoclonal antibody against 6-acetylmorphine protects female mice offspring from adverse behavioral effects induced by prenatal heroin exposure.
Journal of Pharmacology and Experimental Therapeutics.
ISSN 0022-3565.
368(1),
s. 106–115.
doi: 10.1124/jpet.118.251504.
Fulltekst i vitenarkiv
Bergh, Marianne Skov-Skov; Bogen, Inger Lise; Wilson, Steven Ray Haakon & Øiestad, Åse Marit Leere
(2018).
Addressing the Fentanyl Analogue Epidemic by Multiplex UHPLC-MS/MS Analysis of Whole Blood.
Therapeutic Drug Monitoring.
ISSN 0163-4356.
40(6),
s. 738–748.
doi: 10.1097/FTD.0000000000000564.
Bergh, Marianne Skov-Skov; Bogen, Inger Lise; Andersen, Jannike Mørch; Øiestad, Åse Marit Leere & Berg, Thomas
(2017).
Determination of adrenaline, noradrenaline and corticosterone in rodent blood by ion pair reversed phase UHPLC–MS/MS.
Journal of chromatography. B.
ISSN 1570-0232.
1072,
s. 161–172.
doi: 10.1016/j.jchromb.2017.11.016.
Vevelstad, Merete; Øiestad, Elisabeth Leere; Nerem, Elisabeth; Arnestad, Marianne & Bogen, Inger Lise
(2017).
Studies on para-methoxymethamphetamine (PMMA) metabolite pattern and influence of CYP2D6 genetics in human liver microsomes and authentic samples from fatal PMMA intoxications.
Drug Metabolism And Disposition.
ISSN 0090-9556.
45(12),
s. 1326–1335.
doi: 10.1124/dmd.117.077263.
Berntsen, Hanne Friis; Bogen, Inger Lise; Wigestrand, Mattis B; Fonnum, Frode; Walaas, S. Ivar & Moldes-Anaya, Angel
(2017).
The fungal neurotoxin penitrem A induces the production of reactive oxygen species in human neutrophils at submicromolar concentrations.
Toxicology.
ISSN 0300-483X.
392,
s. 64–70.
doi: 10.1016/j.tox.2017.10.008.
Vis sammendrag
Penitrem A is a fungal neurotoxin that recurrently causes intoxication in animals, and occasionally also in humans. We have previously reported that penitrem A induced the production of reactive oxygen species (ROS) in rat cerebellar granule cells, opening for a new mechanism of action for the neurotoxin. The aim of this study was to examine the potential of penitrem A to induce ROS production in isolated human neutrophil granulocytes, and to study possible mechanisms involved.
Penitrem A significantly increased the production of ROS in human neutrophils at concentrations as low as 0.25 μM (40% increase over basal levels), as measured with the DCF fluorescence assay. The EC50 determined for the production of ROS by penitrem A was 3.8 μM. The maximal increase in ROS production was approximately 330% over basal levels at a concentration of 12.5 μM. ROS formation was significantly inhibited by the antioxidant vitamin E (50 μM), the intracellular Ca+2 chelator BAPTA-AM (5 μM), the mitogen activated protein kinase kinase (MEK) 1/2 and 5 inhibitor U0126 (1 and 10 μM), the p38 mitogen activated protein kinase (MAPK) inhibitor SB203580 (1 μM), the c-Jun amino-terminal kinase (JNK) inhibitor SP600125 (10 μM), and the calcineurin inhibitors FK-506 and cyclosporine A (1.5 and 0.5 μM, respectively).
These finding suggest that penitrem A is able to induce an increase in ROS production in neutrophils via the activation of several MAPK-signalling pathways. We suggest that this increase may partly explain the pathophysiology generated by penitrem A neuromycotoxicosis in both humans and animals.
Berntsen, Hanne Friis; Fonnum, Frode; Walaas, S. Ivar & Bogen, Inger Lise
(2016).
Low-chlorinated non-dioxin-like polychlorinated biphenyls present in blood and breast milk induce higher levels of reactive oxygen species in neutrophil granulocytes than high-chlorinated congeners.
Basic & Clinical Pharmacology & Toxicology.
ISSN 1742-7835.
119(6),
s. 588–597.
doi: 10.1111/bcpt.12620.
Bergh, Marianne Skov-Skov; Bogen, Inger Lise; Lundanes, Elsa & Øiestad, Åse Marit Leere
(2016).
Validated methods for determination of neurotransmitters and metabolites in rodent brain tissue and extracellular fluid by reversed phase UHPLC-MS/MS.
Journal of chromatography. B.
ISSN 1570-0232.
1028,
s. 120–129.
doi: 10.1016/j.jchromb.2016.06.011.
Kvello, Anne Marte Sjursen; Andersen, Jannike Mørch; Øiestad, Elisabeth Leere; Mørland, Jørg & Bogen, Inger Lise
(2016).
Pharmacological Effects of a Monoclonal Antibody against
6-Monoacetylmorphine upon Heroin-Induced Locomotor Activity
and Pharmacokinetics in Mice.
Journal of Pharmacology and Experimental Therapeutics.
ISSN 0022-3565.
358(2),
s. 181–189.
doi: 10.1124/jpet.116.233510.
Vevelstad, Merete; Øiestad, Elisabeth Leere; Bremer, Sara; Bogen, Inger Lise; Zackrisson, Anna-Lena & Arnestad, Marianne
(2016).
Is toxicity of PMMA (paramethoxymethamphetamine) associated with cytochrome P450 pharmacogenetics? Forensic Science International.
ISSN 0379-0738.
261,
s. 137–147.
doi: 10.1016/j.forsciint.2016.02.027.
Viluksela, Matti; Heikkinen, Päivi; van der Ven, Leo T.M.; Rendel, Filip; Roos, Robert & Esteban, Javier
[Vis alle 29 forfattere av denne artikkelen](2014).
Toxicological profile of ultrapure 2,2′,3,4,4′,5,5′- heptachlorbiphenyl (PCB 180) in adult rats.
PLOS ONE.
ISSN 1932-6203.
9:e0104639(8).
doi: 10.1371/journal.pone.0104639.
Breivik, Torbjørn; Bogen, Inger Lise; Haug, Kristin Huse; Fonnum, Frode; Opstad, Per Kristian & Eide, Dag Markus
(2014).
Effects of long-term exposure of 3,4-methylenedioxymethamphetamine (MDMA; "ecstasy") on neuronal transmitter transport, brain immuno-regulatory systems and progression of experimental periodontitis in rats.
Neurochemistry International.
ISSN 0197-0186.
72,
s. 30–36.
doi: 10.1016/j.neuint.2014.04.002.
Bogen, Inger Lise; Boix, Fernando; Nerem, Elisabeth; Mørland, Jørg & Andersen, Jannike Mørch
(2014).
A monoclonal antibody specific for 6-monoacetylmorphine reduces acute heroin effects in mice.
Journal of Pharmacology and Experimental Therapeutics.
ISSN 0022-3565.
349(3),
s. 568–576.
doi: 10.1124/jpet.113.212035.
Berntsen, H F; Wigestrand, Mattis Brænne; Bogen, Inger Lise; Fonnum, Frode; Walaas, S. Ivar & Moldes-Anaya, Angel
(2013).
Mechanisms of penitrem-induced cerebellar granule neuron death in vitro: Possible involvement of GABA(A) receptors and oxidative processes.
Neurotoxicology.
ISSN 0161-813X.
35,
s. 129–136.
doi: 10.1016/j.neuro.2013.01.004.
Wendt, Anna; Speidel, Dina; Danielsson, Anders; Esguerra, Jonathan L S; Bogen, Inger Lise & Walaas, S. Ivar
[Vis alle 8 forfattere av denne artikkelen](2012).
Synapsins I and II Are Not Required for Insulin Secretion from Mouse Pancreatic beta-cells.
Endocrinology.
ISSN 0013-7227.
153(5),
s. 2112–2119.
doi: 10.1210/en.2011-1702.
Bogen, Inger Lise; Jensen, Vidar; Hvalby, Øyvind Christian & Walaas, S. Ivar
(2009).
Synapsin-dependent development of glutamatergic vesicles and presynaptic plasticity in postnatal mouse brain.
Neuroscience.
ISSN 0306-4522.
158(1),
s. 231–241.
doi: 10.1016/j.neuroscience.2008.05.055.
Bogen, Inger Lise; Haug, Kristin Huse; Roberg, Bjørg Åse; Fonnum, Frode & Walaas, S. Ivar
(2009).
The importance of synapsin I and II for neurotransmitter levels and vesicular storage in cholinergic, glutamatergic and GABAergic nerve terminals.
Neurochemistry International.
ISSN 0197-0186.
55(1-3),
s. 13–21.
doi: 10.1016/j.neuint.2009.02.006.
Bogen, IL; Risa, Øystein; Risa, O; Haug, KH; Sonnewald, Ursula & Fonnum, F
[Vis alle 7 forfattere av denne artikkelen](2008).
Distinct changes in neuronal and astrocytic amino acid neurotransmitter metabolism in mice with reduced numbers of synaptic vesicles.
Journal of Neurochemistry.
ISSN 0022-3042.
105(6),
s. 2524–2534.
doi: 10.1111/j.1471-4159.2008.05344.x.
Bogen, Inger Lise; Boulland, Jean-Luc; Mariussen, Espen; Wright, Marianne; Fonnum, Frode & Kao, H.T
[Vis alle 7 forfattere av denne artikkelen](2006).
Absence of synapsin I and II is accompanied by decreases in vesicular transport of specific neurotransmitters.
Journal of Neurochemistry.
ISSN 0022-3042.
96,
s. 1458–1466.
doi: 10.1111/j.1471-4159.2005.03636.
Vis sammendrag
Studies of synapsin-deficient mice have shown decreases in the number of synaptic vesicles but knowledge about the consequences of this decrease, and which classes of vesicles are being affected, has been lacking. In this study, glutamatergic, GABAergic and dopaminergic transport has been analysed in animals where the genes encoding synapsin I and II were inactivated. The levels of the vesicular glutamate transporter (VGLUT) 1, VGLUT2 and the vesicular GABA transporter (VGAT) were decreased by approximately 40% in adult forebrain from mice devoid of synapsin I and II, while vesicular monoamine transporter (VMAT) 2 and VGLUT3 were present in unchanged amounts compared with wild-type mice. Functional studies on synaptic vesicles showed that the vesicular uptake of glutamate and GABA was decreased by 41 and 23%, respectively, while uptake of dopamine was unaffected by the lack of synapsin I and II. Double-labelling studies showed that VGLUT1 and VGLUT2 colocalized fully with synapsin I and/or II in the hippocampus and neostriatum, respectively. VGAT showed partial colocalization, while VGLUT3 and VMAT2 did not colocalize with either synapsin I or II in the brain areas studied. In conclusion, distinct vesicular transporters show a variable degree of colocalization with synapsin proteins and, hence, distinct sensitivities to inactivation of the genes encoding synapsin I and II.
Hallberg, Olof Ehlers; Bogen, Inger Lise; Reistad, Trine; Haug, Kristin Huse; Wright, Marianne & Fonnum, Frode
(2006).
Differential development of vesicular glutamate transporters in brain: An in vitro study of cerebellar granule cells.
Neurochemistry International.
ISSN 0197-0186.s. 579–585.
doi: 10.1016/j.neuint.2005.12.027.
Vis sammendrag
The cerebellar granule cells have been extensively used for studies on metabolism, neurotransmission and neurotoxicology, since they can easily be grown in cultures. However, knowledge about the development of different proteins essential for synaptic transmission in these cells is lacking. This study has characterized the developmental profiles of the vesicular glutamate transporters (VGLUTs) and the synaptic vesicle proteins synapsins and synaptophysin in cerebellar granule cells and in co-cultures containing both granule cells and astrocytes. The protein levels of VGLUT2 decreased by approximately 70% from days 2 to 7 in vitro, whereas the levels of VGLUT1 increased by approximately 95%. Protein levels of synapsin I, synapsin IIIa and synaptophysin showed a developmental pattern similar to VGLUT1 while synapsin II and VGLUT3 were absent. The mRNA expressions of VGLUT1 and VGLUT2 were in accordance with the protein levels. The results indicate both that cerebellar granule cells are mature at approximately 7 days in vitro, and that the up-regulation of VGLUT1 and down-regulation of VGLUT2 in cerebellar granule cells are both independent of surrounding astrocytes and neuronal input. The results of this study are discussed in relation to general developmental profiles of VGLUTs in other brain regions
Gundersen, Yngvar; Vaagenes, Per; Thrane, Ingjerd; Bogen, Inger Lise; Haug, Kristin Huse & Reistad, Trine
(2005).
Response of circulating immune cells to major gunshot injury, haemorrhage, and acute surgery.
Injury.
ISSN 0020-1383.
36(8),
s. 949–955.
doi: 10.1016/j.injury.2004.09.021.
Fulltekst i vitenarkiv
Haug, Kristin Huse; Bogen, Inger Lise; Osmundsen, Harald; Walaas, Sven Ivar & Fonnum, Frode
(2005).
Effects on cholinergic markers in rat brain and blood after short and prolonged administration of Donepezil.
Neurochemical Research.
ISSN 0364-3190.
30(12),
s. 1511–1520.
Gylterud, Simen; Bogen, Inger Lise; Walaas, Sven Ivar; Storm-Mathisen, Jon & Bergersen, Linda Hildegard
(2005).
Ultrastructural quantification of glutamate receptors at excitatory synapses in hippocampus of synapsin I+II double knock-out mice.
Neuroscience.
ISSN 0306-4522.
136,
s. 769–777.
Myhre, Oddvar; Sterri, Sigrun Hanne; Bogen, Inger Lise & Fonnum, Frode
(2004).
Erk1/2 phosphorylation and reactive oxygen species formation via nitric oxide and Akt-1/Raf-1 crosstalk in cultured rat cerebellar granule cells exposed to the organic solvent 1,2,4-trimethylcyclohexane.
Toxicological Sciences.
ISSN 1096-6080.
80(2),
s. 296–303.
doi: 10.1093/toxsci/kfh166.
Bogen, Inger Lise; Haug, Kristin Huse; Myhre, Oddvar & Fonnum, Frode
(2003).
Short- and long-term effects of MDMA ("ecstasy") on synaptosomal and vesicular uptake of neurotransmitters in vitro and ex vivo.
Neurochemistry International.
ISSN 0197-0186.
43(4-5),
s. 393–400.
doi: 10.1016/S0197-0186(03)00027-5.
Kogler, Stian; Aizenshtadt, Aleksandra; Harrison, Sean Philip; Sullivan, Gareth John; Lundanes, Elsa & Bogen, Inger Lise
[Vis alle 9 forfattere av denne artikkelen](2023).
Organ-in-a-column – automated on-line incubation, sampling, and LC-MS analysis for measurement of drug metabolites.
Skottvoll, Frøydis Sved; Aizenshtadt, Aleksandra; Harrison, Sean; Krauss, Stefan Johannes Karl; Sullivan, Gareth & Bogen, Inger Lise
[Vis alle 9 forfattere av denne artikkelen](2022).
Organ-in-a-column - On-line incubation and LC-MS-analysis for drug metabolism studies.
Boger, Ida Caroline Sneis; Nerem, Elisabeth; Skottvoll, Frøydis Sved; Harrison, Sean; Sullivan, Gareth & Lundanes, Elsa
[Vis alle 8 forfattere av denne artikkelen](2020).
Determination of liver organoid-induced drug metabolites using liquid chromatography-mass spectrometry (LC-MS)”.
Vis sammendrag
Liver organoids are three-dimensional tissue models typically derived from adult and human induced pluripotent stem cells. They are intended to e.g. represent the physiological functions of a patient´s liver [1]. The liver is the main metabolizing organ in the human body [2]; thus, an important application of liver organoids is to map drug metabolism in vitro as liver organoids can be more patient-specific compared to traditional biomaterials, e.g. human liver microsomes (HLM). The aim of this study is to explore liver organoid drug metabolism in vitro using liquid chromatography-mass spectrometry (LC-MS). To establish a standardized conventional approach for metabolism studies for later comparison with organoids, heroin metabolism studies in HLMs and S9 fraction were carried out. Quantification of model substance heroin and its well-known metabolites, 6-Monoacetylmorphine (6-MAM) and morphine, was done using UHPLC-MS/MS. The heroin metabolism method was miniaturized to make it transferable to the small organoid samples. This was done by decreasing the microsome amount as much as possible while still observing heroin metabolism (HLM amount decreased from 0.2 mg to 0.01 mg; Heroin concentration was decreased from 10 µM to 0.1 µM). This miniaturized experiment was also transferred to S9 fraction Preliminary experiments with organoids showed that the organoids metabolize heroin to 6-MAM and morphine, as well as phase II biotransformation metabolites morphine-3-glucuronide and morphine-6-glucuronide. These results show that the studied organoids have metabolizing properties. Taken together, LC-MS can be a valuable tool for studying metabolism properties of organoids.
Kongstorp, Mette; Andersen, Jannike Mørch; Bogen, Inger Lise & Stiris, Tom
(2019).
Prenatal exposure to opioid maintenance treatment (OMT) reduces cognitive performance in young adult rats.
Liver organoids are three-dimensional tissue models typically derived from adult and human induced pluripotent stem cells. They are intended to e.g. represent the physiological functions of a patient´s liver [1]. The liver is the main metabolizing organ in the human body [2]; thus, an important application of liver organoids is to map drug metabolism in vitro as liver organoids can be more patient-specific compared to traditional biomaterials, e.g. human liver microsomes (HLM). The aim of this study is to explore liver organoid drug metabolism in vitro using liquid chromatography-mass spectrometry (LC-MS). To establish a standardized conventional approach for metabolism studies for later comparison with organoids, heroin metabolism studies in HLMs were carried out. Quantification of model substance heroin and its well-known metabolites, 6-Monoacetylmorphine (6-MAM) and morphine, was done using UHPLC-MS/MS. The heroin metabolism method was miniaturized to make it transferable to the small organoid samples. This was done by decreasing the microsome amount as much as possible while still observing heroin metabolism (HLM amount decreased from 0.2 mg to 0.01 mg; Heroin concentration was decreased from 10 µM to 0.1 µM). Preliminary experiments with organoids showed that the organoids metabolize heroin to 6-MAM and morphine, as well as phase II biotransformation metabolites morphine-3-glucuronide and morphine-6-glucuronide. These results show that the studied organoids have metabolizing properties. We are also exploring sensitive nano-LC-MS (traditionally used for proteomics) for studying organoid drug metabolism. Using a 50 µm (I.D) x 5 cm C18 pre-column and a 50 µm (I.D) x 12 cm C18 analytical column, coupled up with a triple quadrupole MS (QMS) using electrospray ionization (ESI), a LoD of 0.7 fg heroin was achieved. The LoD of the nano-LC-MS method gives a good foundation for future work with detection of trace liver-organoid induced drug metabolites. Taken together, LC-MS can be a valuable tool for studying metabolism properties of organoids.
[1] Eisenstein, M., Organoids: the body builders, Nature Methods, (2018) 19.
[2] Brandon, E. F., Raap, C. D., Meijerman, I., Beijnen, J. H., Schellens, J. H., An update on in vitro test methods in human hepatic drug biotransformation research: pros and cons, Toxicology and applied pharmacology, (2003) 233-246
Liquid chromatography coupled to mass spectrometry (LC-MS) is an analytical tool used to separate and measure the mass (m/z) and concentration of compounds in matrices like water, blood, and tumors. Drugs, metabolites and proteins are all measurable with LC-MS. Unfortunately, the sample is diluted during chromatography, hurting the sensitivity. Thus, the limited sample volumes and low abundant compounds of organ-on-a-chip and organoids demands miniaturization of LC-MS to enable measurements of high sensitivity.
By narrowing the inner diameter (ID) of the separation column, the sample dilution is vastly reduced, adding to sensitivity, e.g. enabling analysis of limited sample volumes and low abundant compounds of the organ-on-a-chip and organoids. The bioanalytical group at the Department of Chemistry (UiO) is currently developing miniaturized systems for proteins and small molecules for assessing the properties of liver organoids. These efforts will be expanded to testing islet and adipose organoids, and will also be incorporated in to on-line systems, e.g. organ on a chip systems. We believe that LC-MS and organoid analysis will be an increasingly natural partnering.
Kongstorp, Mette; Andersen, Jannike Mørch & Bogen, Inger Lise
(2018).
Prenatal exposure to opioid maintenance treatment (OMT) reduced cognitive performance in young adult rats.
Kongstorp, Mette; Andersen, Jannike Mørch & Bogen, Inger Lise
(2018).
The effect on neonatal rat offspring born to dams exposed to methadone or buprenorphine during pregnancy.
Kongstorp, Mette; Andersen, Jannike Mørch & Bogen, Inger Lise
(2017).
Neurobiological development in rat pups exposed to methadone or buprenorphine during gestation.
Bogen, Inger Lise; Boix, Fernando; Nerem, Elisabeth; Mørland, Jørg & Andersen, Jannike Mørch
(2013).
Prekliniske studier av en heroin-vaksine.
Bogen, Inger Lise; Boix, Fernando; Nerem, Elisabeth; Mørland, Jørg & Andersen, Jannike Mørch
(2013).
Preklinisk studie av en passiv heroinvaksine.
Bogen, Inger Lise; Andersen, Jannike Mørch; Boix, Fernando; Nerem, Elisabeth; Normann, Per Trygve & Mørland, Jørg
(2012).
Heroin abuse and addiction: Heroin metabolism as a key player and possible target for new pharmacotherapy.
The liver is the main metabolizing organ in the human body, and several in vitro models have been developed to resemble hepatic drug metabolism. However, commonly used in vitro models such as human liver microsomes (HLMs), S9 fraction, and hepatocytes lack the complexity of the corresponding in vivo tissue, which limits the in vivo resemblance. Liver organoids are three-dimensional tissue models typically derived from induced pluripotent stem cells (iPSCs) and are intended to recapitulate physiological functions of the human liver. Liver spheroids are similar 3D culture systems but lack the multicellularity that characterizes the organoids. Few studies have investigated drug metabolism in liver organoids- and spheroids. The aim of this study was, therefore, to explore liver organoid- and spheroid drug metabolism using liquid chromatography-mass spectrometry (LC-MS) as the liver organoids- and spheroids can be more representative to the in vivo situation. Additionally, an LC-MS method was to be established at the Department of Chemistry for drug metabolite detection. Drug incubated samples were analyzed using a validated LC-MS method at the Department of Forensic Sciences or by the LC-MS method established at the Department of Chemistry. Heroin was chosen as the model drug, and heroin metabolism studies were initially carried out in HLMs to establish a standardized conventional approach for later drug metabolism studies in organoids and spheroids. The concentrations of heroin and its phase I metabolites, 6-monoacetylmorphine (6-MAM) and morphine were measured over time (20 minutes). The heroin metabolism method was downscaled by decreasing the HLM concentration (from 2 mg/mL to 0.1 mg/mL) and the heroin concentration (from 1 μM to 0.1 μM). The downscaled method was applied to the S9 fraction and later executed with the addition of exogenous cofactors to look for the phase II metabolites morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G). With the addition of cofactors, M3G could be quantified in the S9 fraction. Preliminary heroin metabolism studies in iPSC derived liver organoids showed that the organoids metabolized heroin (1 μM) to 6-MAM and morphine. The phase II metabolites M3G and M6G were also identified but below the limit of quantification (LOQ). Both the phase I- and phase II metabolites were detected above LOQ in liver spheroid samples after incubation in 10 M heroin. To sum up, the detection of heroin drug metabolites using LC-MS showed that the liver organoids- and spheroids had heroin metabolizing properties. Additionally, LC-MS proved to be a valuable tool for detection of liver organoid induced drug metabolites.
