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Translational Non-Invasive Metabolic Studies towards Novel Treatments of Chronic Hepatic Encephalopathy in Developing Brain, from 3D Organotypic Brain Cell Cultures to the In vivo Rat and Human Brain

English title Translational Non-Invasive Metabolic Studies towards Novel Treatments of Chronic Hepatic Encephalopathy in Developing Brain, from 3D Organotypic Brain Cell Cultures to the In vivo Rat and Human Brain
Applicant Cudalbu Cristina
Number 173222
Funding scheme Project funding
Research institution Centre d'Imagerie BioMédicale CIBM EPFL - SB - IPSB - LIFMET
Institution of higher education EPF Lausanne - EPFL
Main discipline Paediatrics
Start/End 01.04.2017 - 31.07.2021
Approved amount 518'206.00
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All Disciplines (3)

Discipline
Paediatrics
Biophysics
Pathophysiology

Keywords (8)

Hepatic Encephalopathy; in vivo Magnetic Resonance Spectroscopy / Imaging; creatine; brain energy metabolism; osmoregulation; brain metabolism; brain edema; Chronic liver disease

Lay Summary (French)

Lead
L’encéphalopathie hépatique est une complication neurologique grave due à l’insuffisance hépatique chronique, affectant à la fois les adultes et les enfants. L’atrésie des voies biliaires est la cause la plus fréquente de l’insuffisance hépatique chronique chez les enfants et est également l'indication la plus fréquente de transplantation hépatique pédiatrique. L'augmentation de la concentration cérébrale de l'ammoniaque est considérée à être la cause majeure de l’encéphalopathie hépatique mais sa relation avec les lésions neurologiques est mal comprise. Le rôle des acides biliaires est aussi mal connu.Le projet apporte sa contribution a ce domaine de recherche
Lay summary

Chez les adultes ayant une l’insuffisance hépatique chronique il est bien accepte que l’hyperammonémie va générer une augmentation de la glutamine cérébrale et une perturbation osmotique secondaire qui va entrainer un œdème cérébrale diffus.  Bien que dans l'âge adulte l’encéphalopathie hépatique est en grande partie réversible lors du retour de l'ammoniac sérique à des valeurs normales, l’hyperammonémie est associée à des dommages irréversibles au système nerveux central chez l'enfant. Des déficits résiduels similaires n'ont pas été décrits chez les patients adultes après l’insuffisance hépatique chronique. La compréhension des mécanismes moléculaires qui sont à la base de ces déficits neurocognitifs est importante pour finalement être en mesure de proposer des stratégies de neuroprotection pour ces enfants dans le but d'améliorer leurs résultats à long terme.

En se basant sur nos résultats préliminaires, nous émettons l'hypothèse que des différences dans l'osmoregulation, les neurotransmetteurs, antioxydants et les métabolites énergétiques sont à la base des différences neurologiques observés dans le cerveau mature et en développement durant une l’insuffisance hépatique chronique. Cette susceptibilité différentielle peut être liée à la déficience secondaire en Cr et à la toxicité combinée de l’ammoniac avec les acides biliaires.

Le projet est divisé en plusieurs phases. Dans une première phase nous allons étudier les différences entre le cerveau en développement et celui mature pendant l’insuffisance hépatique chronique en réalisant des mesures in vitro et in vivo et en utilisant l’imagerie et la spectroscopie de résonance magnétique. Dans une deuxième partie nous allons évaluer le rôle neuroprotecteur de la créatine dans l’insuffisance hépatique chronique. La dernière partie de notre projet implique l'étude métabolique du cerveau chez les patient pédiatriques atteints d'une insuffisance hépatique chronique.  

Direct link to Lay Summary Last update: 05.04.2017

Responsible applicant and co-applicants

Employees

Publications

Publication
Probiotics combined with rifaximin influence the neurometabolic changes in a rat model of type C HE
Flatt Emmanuelle, McLin Valérie A., Braissant Olivier, Pierzchala Katarzyna, Mastromarino Paola, Mitrea Stefanita-Octavian, Sessa Dario, Gruetter Rolf, Cudalbu Cristina (2021), Probiotics combined with rifaximin influence the neurometabolic changes in a rat model of type C HE, in Scientific Reports, 11(1), 17988-17988.
In vivo macromolecule signals in rat brain 1 H‐MR spectra at 9.4T: Parametrization, spline baseline estimation, and T 2 relaxation times
Simicic Dunja, Rackayova Veronika, Xin Lijing, Tkáč Ivan, Borbath Tamas, Starcuk Zenon, Starcukova Jana, Lanz Bernard, Cudalbu Cristina (2021), In vivo macromolecule signals in rat brain 1 H‐MR spectra at 9.4T: Parametrization, spline baseline estimation, and T 2 relaxation times, in Magnetic Resonance in Medicine, 86(5), 2384-2401.
2021 ISHEN guidelines on animal models of hepatic encephalopathy
DeMorrow Sharon, Cudalbu Cristina, Davies Nathan, Jayakumar Arumugam R., Rose Christopher F. (2021), 2021 ISHEN guidelines on animal models of hepatic encephalopathy, in Liver International, 41(7), 1474-1488.
Probiotics improve the neurometabolic profile of rats with chronic cholestatic liver disease
RačkayováVeronika, Flatt Emmanuelle, BraissantOlivier, GrosseJocelyn, CapobiancoDaniela, MastromarinoPaola, McMillinMatthew, DeMorrow Sharon, McLinValérie A, CudalbuCristina (2021), Probiotics improve the neurometabolic profile of rats with chronic cholestatic liver disease, in Sci Rep, 11(1), 2269.
Late post-natal neurometabolic development in healthy male rats using 1 H and 31 P magnetic resonance spectroscopy
RačkayováVeronika, SimicicDunja, Donati Guillaume, BraissantOlivier, GruetterRolf, McLin Valérie A, CudalbuCristina (2021), Late post-natal neurometabolic development in healthy male rats using 1 H and 31 P magnetic resonance spectroscopy, in J Neurochem , XX.
Longitudinal osmotic and neurometabolic changes in young rats with chronic cholestatic liver disease
Rackayova Veronika, Braissant Olivier, Rougemont Anne-Laure, Cudalbu Cristina, McLin Valérie A. (2020), Longitudinal osmotic and neurometabolic changes in young rats with chronic cholestatic liver disease, in Scientific Reports, 10(1), 7536-7536.
B 0 shimming for in vivo magnetic resonance spectroscopy: Experts' consensus recommendations
Juchem Christoph, Cudalbu Cristina, Graaf Robin A., Gruetter Rolf, Henning Anke, Hetherington Hoby P., Boer Vincent O. (2020), B 0 shimming for in vivo magnetic resonance spectroscopy: Experts' consensus recommendations, in NMR Biomed, e4350.
Contribution of macromolecules to brain 1 H MR spectra: Experts' consensus recommendations
Cudalbu Cristina, Behar Kevin L., Bhattacharyya Pallab K., Bogner Wolfgang, Borbath Tamas, Graaf Robin A., Gruetter Rolf, Henning Anke, Juchem Christoph, Kreis Roland, Lee Phil, Lei Hongxia, Marjańska Małgorzata, Mekle Ralf, Murali‐Manohar Saipavitra, Považan Michal, Rackayová Veronika, Simicic Dunja, Slotboom Johannes, Soher Brian J., Zenon Starčuk Jr., Starčuková Jana, Tkáč Ivan, Williams Stephen, Wilson Martin, Wright Andrew Martin, Xin Lijing, Mlynárik Vladimír (2020), Contribution of macromolecules to brain 1 H MR spectra: Experts' consensus recommendations, in NMR Biomed, e4393.
Magnetic resonance spectroscopy in the rodent brain: Experts' consensus recommendations
Lanz Bernard, Abaei Alireza, Braissant Olivier, Choi In‐Young, Cudalbu Cristina, Henry Pierre‐Gilles, Gruetter Rolf, Kara Firat, Kantarci Kejal, Lee Phil, Lutz Norbert W., Marjańska Małgorzata, Mlynárik Vladimír, Rasche Volker, Xin Lijing, Valette Julien, Behar Kevin, Boumezbeur Fawzi, Deelchand Dinesh Kumar, Dreher Wolfgang, Klaunberg Brenda A., Ligneul Clemence, Lindquist Diana M., Öz Gülin, Tkáč Ivan, Biomed NMR (2020), Magnetic resonance spectroscopy in the rodent brain: Experts' consensus recommendations, in NMR Biomed, e4325.
Terminology and concepts for the characterization of in vivo MR spectroscopy methods and MR spectra: Background and experts' consensus recommendations
Kreis Roland, Boer Vincent, Choi In-Young, Cudalbu Cristina, Graaf Robin A., Gasparovic Charles, Heerschap Arend, Krššák Martin, Lanz Bernard, Maudsley Andrew A., Meyerspeer Martin, Near Jamie, Öz Gülin, Posse Stefan, Slotboom Johannes, Terpstra Melissa, Tkáč Ivan, Wilson Martin, Bogner Wolfgang (2020), Terminology and concepts for the characterization of in vivo MR spectroscopy methods and MR spectra: Background and experts' consensus recommendations, in {NMR} in Biomedicine, n/a.
Longitudinal neurometabolic changes in the hippocampus of a rat model of chronic hepatic encephalopathy
Braissant Olivier, Rackayová Veronika, Pierzchala Katarzyna, Grosse Jocelyn, McLin Valérie, Cudalbu Cristina (2019), Longitudinal neurometabolic changes in the hippocampus of a rat model of chronic hepatic encephalopathy, in Journal of Hepatology, 71(3), 505-515.
Brain Edema in Chronic Hepatic Encephalopathy
Cudalbu Cristina, Taylor-Robinson Simon D. (2019), Brain Edema in Chronic Hepatic Encephalopathy, in Journal of Clinical and Experimental Hepatology, 9(3), 362-382.

Datasets

MR Spectra from rat hippocampus with LCModel quantification and the corresponding basis set

Author Simicic, Dunja; Cudalbu, Cristina
Publication date 23.06.2021
Persistent Identifier (PID) https://doi.org/10.5281/zenodo.3904443
Repository Spectra_hippocampus(rat)_TE02.rar
Abstract
This folder contains the LCModel quantifications of spectra acquired in hippocampus from 7 rats. The spectra were quntified using six different DKNTMN (spline stiffness) values (0.1, 0.25, 0.4, 0.5, 1, 5). In the folder Control_files_Basis_set you can find all the control files used in this quantification along with the corresponding basis set (metabolites/simulated using NMRScopeB from jMRUI and in vivo parameters + full MM spectrum).

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
The 19th International Society of Hepatic Encephalopathy and Nitrogen Metabolism, Symposium- Talk given at a conference Central Nervous System Oxidative Stress and Inflammation in a rat model of Type C Hepatic Encephalopathy – brothers in arms? 06.10.2021 online, Italy Pierzchala Katarzyna; McLin Valérie Anne; Simicic Dunja; Braissant Olivier; Cudalbu Cristina;
19th International Society of Hepatic Encephalopathy and Nitrogen Metabolism, Symposium Individual talk Brain oedema in persistent hepatic encephalopathy 06.10.2021 online, Italy Cudalbu Cristina;
ISMRM Annual Meeting August 2021 Poster Protective rCreatine in chronic hepatic encephalopathy developing brain: in vivo longitudinal 1H and 31P-MRS study 15.05.2021 online, Switzerland Braissant Olivier; Simicic Dunja; McLin Valérie Anne; Cudalbu Cristina; Pierzchala Katarzyna;
ISMRM Annual Meeting August 2020 Talk given at a conference Neurometabolism in children with chronic liver disease or portosystemic shunting: a 1H-MRS/MRI study at 7T 08.08.2020 online, France McLin Valérie Anne; Cudalbu Cristina;
CIBM BREAKFAST & SCIENCE SEMINARS Individual talk Translational Non-Invasive Neurometabolic Studies during Chronic Hepatic Encephalopathy in Developing Brain, from the In vivo Rat to the Human Brain 31.03.2020 https://cibm.ch/event-cibm-breakfast-science-seminar-3-march-31-2020/, Switzerland Simicic Dunja; Cudalbu Cristina; McLin Valérie Anne; Pierzchala Katarzyna; Rackayová Veronika; Braissant Olivier;
International Society for Hepatic Encephalopathy and Nitrogen Metabolism (ISHEN) Poster Brain regional susceptibility to Oxidative Stress in a rat model of Chronic Hepatic Encephalopathy: in-vivo 1H MRS, ex-vivo ESR spectroscopy and histology findings 12.09.2019 Williamsburg, United States of America Cudalbu Cristina; Braissant Olivier; McLin Valérie Anne; Simicic Dunja; Rackayová Veronika; Pierzchala Katarzyna;
International Society for Hepatic Encephalopathy and Nitrogen Metabolism (ISHEN) Poster In vivo longitudinal 1H MRS study of hippocampal, cereberal and striatal metabolic changes in the adult brain using an animal model of Chronic Hepatic Encephalopathy 12.09.2019 Williamsburg, United States of America Cudalbu Cristina; Pierzchala Katarzyna; Simicic Dunja; Braissant Olivier; Rackayová Veronika; McLin Valérie Anne;
International Society for Hepatic Encephalopathy and Nitrogen Metabolism (ISHEN) Poster Neurometabolism in grey matter of children with chronic liver disease or portosystemic shunting: a 1H-MRS study at 7T 12.09.2019 Williamsburg, United States of America Cudalbu Cristina; McLin Valérie Anne;
International Society for Hepatic Encephalopathy and Nitrogen Metabolism (ISHEN) Poster Hippocampal and cerebellar astrocytes morphological alterations in a rat model of chronic hepatic encephalopathy 12.09.2019 Williamsburg , United States of America Cudalbu Cristina; Simicic Dunja; McLin Valérie Anne; Braissant Olivier; Pierzchala Katarzyna;
Joint Annual Meeting ISMRM-ESMRMB 2018 Poster Late post-natal brain metabolic changes in healthy rats, a longitudinal in vivo 1H and 31P MRS study 16.06.2019 Paris, France Rackayová Veronika; McLin Valérie Anne; Cudalbu Cristina; Braissant Olivier;
ISMRM 27th Annual Meeting & Exhibition Talk given at a conference Chronic hepatic encephalopathy in early developing brain, neurometabolic changes differ depending on the age of disease onset, in vivo longitudinal 1H MRS study 11.05.2019 Montreal, Canada Braissant Olivier; Rackayová Veronika; McLin Valérie Anne; Cudalbu Cristina;
MRS Workshop 2018-Metabolic Imaging Talk given at a conference Early and profound 1H MRS neurometabolic changes in pup rat with cholestatic liver disease compared to changes in adult rats 03.10.2018 Utrecht, Netherlands Cudalbu Cristina; Rackayová Veronika; McLin Valérie Anne; Braissant Olivier;
MRS workshop 2018- Metabolic imaging Talk given at a conference In vivo longitudinal 1H MRS Study of hippocampal, cerebral and striatal metabolic changes in BDL rats 03.10.2018 Utrecht, Netherlands Cudalbu Cristina; Pierzchala Katarzyna; Simicic Dunja; McLin Valérie Anne; Rackayová Veronika; Braissant Olivier;
Joint Annual Meeting ISMRM-ESMRMB 2018 Poster Protective effect of high creatine diet during chronic hepatic encephalopathy in young rats, an in vivo longitudinal 1H and 31P MRS study 16.06.2018 Paris, France Braissant Olivier; McLin Valérie Anne; Cudalbu Cristina; Rackayová Veronika;


Communication with the public

Communication Title Media Place Year
New media (web, blogs, podcasts, news feeds etc.) MRS Experts’ consensus recommendation published in ‘NMR Biomedicine’ International 2021
Media relations: print media, online media Des chercheurs romands exposent pourquoi le cerveau trinque quand le foie va mal Heidi News International Western Switzerland 2019
Media relations: print media, online media When liver disease affects the brain EPFL actu news International 2019

Associated projects

Number Title Start Funding scheme
201218 Enhanced MR Spectroscopic mapping of brain regional changes in type C hepatic encephalopathy of juvenile rats to develop novel combinatorial treatments 01.10.2021 Project funding
201218 Enhanced MR Spectroscopic mapping of brain regional changes in type C hepatic encephalopathy of juvenile rats to develop novel combinatorial treatments 01.10.2021 Project funding
175778 Cerebral creatine deficiency syndromes: New in vivo AAV approaches to treat SLC6A8 deficiency 01.11.2017 Project funding

Abstract

1.1BackgroundChronic hepatic encephalopathy (HE) is a well-accepted complication of cholestatic, chronic liver disease (CLD) in adults. It is less well understood in children, although many children with cholestatic CLD display an array of neurocognitive deficits. Increase in ammonium (NH4+) delivery to the brain in CLD is thought to be the main culprit. The role of bile acids in these deficits is unknown. It is commonly accepted in adults with CLD that increased central nervous system NH4+ generates a rise in the osmolyte glutamine and a secondary osmotic imbalance, recently shown to be associated with low grade brain edema. These changes might impact brain energy metabolism, but seem to be largely corrected when blood NH4+ declines. However, based on current understanding of infants with urea cycle defects (UCD)-characterized by acute and extreme hyperammonemia (HA), it appears that the developing (infant) brain under HA faces irreversible impairment of brain cell migration and differentiation and ultimately significant brain cell death. Additionally, there is emerging evidence that chronic cholestasis early in life may be associated with long-term neurocognitive and neuromotor deficits. No similar long-term deficits have been described in adult patients. How the infant or child’s brain responds to the metabolic changes of CLD, and how these mechanisms differ from those in adult patients are both unknown. Further, prophylactic or therapeutic measures are needed to prevent or treat the effects of CLD in children. Our previous in vitro measurements showed that NH4+ in developing brain cells leads to cell death, inhibition of axonal growth and generates a secondary creatine (Cr) deficiency, while Cr supplementation shows neuroprotective effects. Our preliminary data also show in vitro that several bile acids occurring in CLD may be toxic to neurons and astrocytes. Using in vivo 1H, 31P,13C MR spectroscopy (MRS) and Diffusion Tensor Imaging (DTI) in a rat model of CLD we have recently shown that rat pups display a distinct brain metabolism from adults. First, we observed a greater glutamine increase, together with the predicted myo-inositol decrease, yet more pronounced brain edema than predicted. Next, pups displayed a stronger decrease in neurotransmitters, and antioxidants and evidence of altered energy metabolism (lactate). Importantly for the proposal, there was a significant decrease in measured Cr, while Cr treatment increased brain N-acetylaspartate levels in rat pups. Cr has multiple essential functions in the central nervous system: it participates in energy metabolism, osmoregulation, and neurotransmission. Therefore, Cr emerges as a likely candidate underlying the differential susceptibility to CLD between adults and pups.1.2Working hypothesisWe hypothesize that differences in osmoregulation, neurotransmitter, antioxidant and energy metabolism underlie the differential susceptibility to CLD-induced neurological changes observed in the mature and developing brain, and that this differential susceptibility may be related to secondary Cr deficiency, and the combined toxicities of bile acids and NH4+.1.3AimsI: Differences between mature and developing brain during CLD: from 3D organotypic brain cell cultures to an in vivo rat modelII: Neuroprotective role of Cr in the adult and developing rat brain with CLD-induced HE.III: Abnormalities in brain Gln, Ins, Cr, Glu and cognitive performance in children with CLD1.4Experimental design and/or methods-We will assess the effects of bile acids with/without NH4+ on 3D brain cell cultures using biochemical and histological methods.-We will analyze the longitudinal progression of 16 brain metabolites non-invasively, in vivo, with a focus on osmoregulation, energy, neurotransmitter, antioxidant metabolites, using 1H MRS in rats and children with CLD. -We will simultaneously measure brain edema and changes in white matter maturation in vivo by DTI in rats with CLD. -We will investigate neuroglial energy metabolism in vivo using 13C MRS and 31P MRS in rats with CLD.-We will analyze the effects of HE using biochemical and histological methods in the brain of the same rats evaluated by 1H, 13C, 31P MRS as well as DTI.-The neuroprotective effects of Cr supplementation will be assessed in vitro against the toxic effects of bile acids with- or without NH4+ exposure, as well as in vivo in pups and adult CLD rats by looking at the effect of Cr supplementation using the approaches described above.1.5Expected values of the proposed projectThe proposed experiments aim to decipher the molecular underpinnings behind the differential vulnerability of the developing vs the mature brain to the insults of CLD using a 3D brain cell culture model to analyze the impact of HA and bile acids. This will be correlated with in vivo studies to analyze the temporal differences between pup and adult rats with CLD by examining in detail the longitudinal variations in osmoregulation, neurotransmission, antioxidants, energy metabolites, white matter maturation and edema. Using this multimodal approach, we will specifically monitor differences in Cr metabolism and analyze the response to Cr supplementation in the diet of CLD animals, thereby testing for the first time the hypothesis that fluctuations in Cr may underlie the age-dependent differential susceptibility to HE, and the potential neuroprotective effect of Cr supplementation. In parallel, we will pave the way to future interventional studies in children, in whom we will analyze the link between neurometabolism and neurocognitive performance in a preliminary study.
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