Publications

Tumour cells can escape antiproliferative pressure by interferon-β through immunoediting of interferon receptor expression
Abstract Type I interferons (IFNs) play a central role not only in innate immunity against viral infection, but also in the antitumour response. Apart from indirect immune-modulatory and anti-angiogenic effects, they have direct impact on cell proliferation. Particularly for cancer arising in the context of chronic inflammation, constant exposure to IFNs may constitute a strong selective pressure during tumour evolution. Expansion of neoplastic subclones or -populations that developed resistance to the antiproliferative effects of IFNs might constitute an important contribution to immunoediting of the cancer cells leading to more aggressive and metastasising disease. Experimental evidence for this development of IFN-insensitivity has been scarce and its molecular mechanism is unclear. In this study we demonstrate that prolonged (six weeks) exposure of cells to IFN-β in vitro reduces their sensitivity to its antiproliferative effects, and that this phenotype was stable for up to four weeks. Furthermore, we observed substantial differences in cellular sensitivity to growth inhibition by IFN-β in a panel of ten different liver cancer cell lines of varying malignity. IFN-resistance was most prominent in a pair of highly dedifferentiated cell lines, and least in cells from well-differentiated tumours, fostering the hypothesis of IFN-driven immunoediting in advanced cancers. In both settings, long-term IFN selection in vitro as well as in dedifferentiated tumour cell lines, we found IFNAR expression to be substantially reduced, suggesting the receptor complex, in particular IFNAR2, to be a sensitive target amenable to immunoediting. Beyond new insights into possible molecular processes in tumour evolution, these findings might prove valuable for the development of biomarkers allowing to stratify tumours for their sensitivity to IFN treatment in the context of patient tailored therapies.
DOI: 10.21203/rs.3.rs-3276415/v1
Authors:
  • Felix Hiebinger
  • Aiste Kudulyte
  • Huanting Chi
  • Sebastian Burbano de Lara
  • Barbara Helm
  • Hendrik Welsch
  • Viet Loan Dao Thi
  • Ursula Klingmüller
  • Marco Binder
https://doi.org/10.21203/rs.3.rs-3276415/v1
2023-08-24
TGF-β1 inhibits cholesterol metabolism in hepatocytes to facilitate cell death, EMT and signals for HSC activation.
Objective: Transforming growth factor-β1 (TGF-β1) plays important roles in chronic liver diseases, including metabolic dysfunction-associated steatotic liver disease (MASLD), which involves various biological processes including dysfunctional cholesterol metabolism contributing to progression to metabolic dysfunction-associated steatohepatitis (MASH) and hepatocellular carcinoma (HCC). However, how TGF-β1 signaling and cholesterol metabolism affects each other in MASLD is yet unknown. Design: Changes in transcription of genes associated with cholesterol metabolism were assessed by RNA-Seq of AML12 cells and mouse primary hepatocytes (MPH) treated with TGF-β1. Functional assays were performed on AML12 cells (untreated, TGF-β1 treated, or subjected to cholesterol enrichment (CE) or depletion (CD)), and on mice injected with adeno-associated virus 8 (AAV8)-Control/TGF-β1. Results: TGF-β1 inhibited mRNA expression of several cholesterol metabolism regulatory genes, including rate-limiting enzymes of cholesterol biosynthesis in AML12 cells, MPHs, and AAV8-TGF- β1-treated mice. Total cholesterol levels in AML12 cells, as well as lipid droplet accumulation in AML12 cells and AAV-treated mice were also reduced. Smad2/3 phosphorylation following 2 h TGF-β1 treatment persisted after CE or CD and was mildly increased following CD, while TGF-β1-mediated AKT phosphorylation (30 min) was inhibited by CE. Furthermore, CE protected AML12 cells from several effects mediated by 72 h incubation with TGF-β1, including EMT, actin polymerization, and apoptosis. CD mimicked the outcome of long term TGF-β1 administration, an effect that was blocked by an inhibitor of the type I TGF-β receptor. Additionally, the supernatant of CE- or CD-treated AML12 cells inhibited or promoted, respectively, the activation of LX-2 hepatic stellate cells. Conclusion: TGF-β1 inhibits cholesterol metabolism while cholesterol attenuates TGF-β1 downstream effects in hepatocytes.
DOI: 10.1101/2023.08.14.552900
Authors:
  • Sai Wang
  • Frederik Link
  • Mei Han
  • Roman Liebe
  • Ye Yao
  • Seddik Hammad
  • Anne Dropmann
  • Roohi Chaudhary
  • Anastasia Asimakopoulos
  • Marinela Krizanac
  • Ralf Weiskirchen
  • Yoav I Henis
  • Marcelo Ehrlich
  • Matthias Ebert
  • Steven Dooley
https://doi.org/10.1101/2023.08.14.552900
2023-08-15
Hepatokine‐based identification of fibrotic NASH and improved risk stratification in a multicentre cohort of NAFLD patients
Abstract Background and Aims The presence of significant liver fibrosis associated with non‐alcoholic steatohepatitis (NASH) is regarded as the major prognostic factor in non‐alcoholic fatty liver disease (NAFLD). Identification of patients at risk for NASH with significant fibrosis is therefore important. Although the established fibrosis score FIB‐4 is suitable to exclude advanced fibrosis, it does not allow the prediction of significant fibrosis in NAFLD patients. We therefore evaluated whether the hepatokine fibroblast growth factor 21 (FGF21), a regulator of glucose and lipid metabolism, might identify ‘at‐risk NASH’ in NAFLD. Methods FGF21 levels were assessed by enzyme‐linked immunosorbent assay in sera from an exploration ( n  = 137) and a validation ( n  = 88) cohort of biopsy‐proven NAFLD patients with different disease activity and fibrosis stages. In addition, we evaluated whether the use of FGF21 could improve risk stratification in NAFLD patients with low (<1.3) or intermediate (1.3–2.67) FIB‐4. Results FGF21 levels could significantly discriminate between NASH and non‐alcoholic fatty liver (NAFL) patients, even in the absence of diabetes. Moreover, patients with NASH and fibrosis ≥F2 showed significantly higher FGF21 levels compared to NAFLD patients without significant fibrosis. Significantly elevated FGF21 levels could even be detected in NAFLD patients with NASH and significant fibrosis despite low or intermediate FIB‐4. Conclusion Serological FGF21 detection might allow the identification of NAFLD patients at risk and improves patient stratification in combination with FIB‐4.
DOI: 10.1111/liv.15686
Authors:
  • Martin Franck
  • Katharina John
  • Sherin Al Aoua
  • Monika Rau
  • Andreas Geier
  • Jörn M. Schattenberg
  • Heiner Wedemeyer
  • Klaus Schulze‐Osthoff
  • Heike Bantel
https://doi.org/10.1111/liv.15686
Liver International2023-08-03
An integrative experimental and computational twin modeling approach to understand clonal dynamics in the normal liver
DOI: 10.1016/j.jhep.2023.05.016
Authors:
  • Dirk Drasdo
  • Jieling Zhao
https://doi.org/10.1016/j.jhep.2023.05.016
Journal of Hepatology2023-08-01
Anti-Cancer Prodrug Cyclophosphamide Exerts Thrombogenic Effects on Human Venous Endothelial Cells Independent of CYP450 Activation—Relevance to Thrombosis
Cancer patients are at a very high risk of serious thrombotic events, often fatal. The causes discussed include the detachment of thrombogenic particles from tumor cells or the adverse effects of chemotherapeutic agents. Cytostatic agents can either act directly on their targets or, in the case of a prodrug approach, require metabolization for their action. Cyclophosphamide (CPA) is a widely used cytostatic drug that requires prodrug activation by cytochrome P450 enzymes (CYP) in the liver. We hypothesize that CPA could induce thrombosis in one of the following ways: (1) damage to endothelial cells (EC) after intra-endothelial metabolization; or (2) direct damage to EC without prior metabolization. In order to investigate this hypothesis, endothelial cells (HUVEC) were treated with CPA in clinically relevant concentrations for up to 8 days. HUVECs were chosen as a model representing the first place of action after intravenous CPA administration. No expression of CYP2B6, CYP3A4, CYP2C9 and CYP2C19 was found in HUVEC, but a weak expression of CYP2C18 was observed. CPA treatment of HUVEC induced DNA damage and a reduced formation of an EC monolayer and caused an increased release of prostacyclin (PGI2) and thromboxane (TXA) associated with a shift of the PGI2/TXA balance to a prothrombotic state. In an in vivo scenario, such processes would promote the risk of thrombus formation.
DOI: 10.3390/cells12151965
Authors:
  • Anne Krüger-Genge
  • Susanne Köhler
  • Markus Laube
  • Vanessa Haileka
  • Sandy Lemm
  • Karolina Majchrzak
  • Sarah Kammerer
  • Christian Schulz
  • Joachim Storsberg
  • Jens Pietzsch
  • Jan-Heiner Küpper
  • Friedrich Jung
https://doi.org/10.3390/cells12151965
Cells2023-08-01
Myostatin is associated with the presence and development of acute-on-chronic liver failure
DOI: 10.1016/j.jhepr.2023.100761
Authors:
  • Astrid Ruiz-Margáin
  • Alessandra Pohlmann
  • Silke Lanzerath
  • Melanie Langheinrich
  • Alejandro Campos-Murguía
  • Berenice M. Román-Calleja
  • Robert Schierwagen
  • Sabine Klein
  • Frank Erhard Uschner
  • Maximilian Joseph Brol
  • Aldo Torre-Delgadillo
  • Nayelli C. Flores-García
  • Michael Praktiknjo
  • Ricardo U. Macías Rodríguez
  • Jonel Trebicka
https://doi.org/10.1016/j.jhepr.2023.100761
JHEP Reports2023-08-01
Development, validation, and prognostic evaluation of a risk score for long-term liver-related outcomes in the general population: a multicohort study
DOI: 10.1016/S0140-6736(23)01174-1
Authors:
  • Miquel Serra-Burriel
  • Adrià Juanola
  • Feliu Serra-Burriel
  • Maja Thiele
  • Isabel Graupera
  • Elisa Pose
  • Guillem Pera
  • Ivica Grgurevic
  • Llorenç Caballeria
  • Salvatore Piano
  • Laurens van Kleef
  • Mathias Reichert
  • Dominique Roulot
  • Juan M Pericàs
  • Jörn M Schattenberg
  • Emmanuel A Tsochatztis
  • Indra Neil Guha
  • Montserrat Garcia-Retortillo
  • Rosario Hernández
  • Jordi Hoyo
  • Matilde Fuentes
  • Carmen Expósito
  • Alba Martínez
  • Patricia Such
  • Anita Madir
  • Sönke Detlefsen
  • Marta Tonon
  • Andrea Martini
  • Ann T Ma
  • Judith Pich
  • Eva Bonfill
  • Marta Juan
  • Anna Soria
  • Marta Carol
  • Jordi Gratacós-Ginès
  • Rosa M Morillas
  • Pere Toran
  • J M Navarrete
  • Antoni Torrejón
  • Céline Fournier
  • Anne Llorca
  • Anita Arslanow
  • Harry J de Koning
  • Fernando Cucchietti
  • Michael Manns
  • Phillip N Newsome
  • Rubén Hernáez
  • Alina Allen
  • Paolo Angeli
  • Robert J de Knegt
  • Tom H Karlsen
  • Peter Galle
  • Vincent Wai-Sun Wong
  • Núria Fabrellas
  • Laurent Castera
  • Aleksander Krag
  • Frank Lammert
  • Patrick S Kamath
  • Pere Ginès
  • Marifé Alvarez
  • Peter Andersen
  • Paolo Angeli
  • Alba Ardèvol
  • Anita Arslanow
  • Luca Beggiato
  • Zahia Ben Abdesselam
  • Lucy Bennett
  • Bajiha Boutouria
  • Alessandra Brocca
  • M. Teresa Broquetas
  • Llorenç Caballeria
  • Valeria Calvino
  • Judith Camacho
  • Aura Capdevila
  • Marta Carol
  • Laurent Castera
  • Marta Cervera
  • Fernando Cucchietti
  • Anna de Fuentes
  • Rob de Knegt
  • Harry J de Koning
  • Sonke Detlefsen
  • Alba Diaz
  • José Diéguez Bande
  • Vanessa Esnault
  • Núria Fabrellas
  • Josep Lluis Falcó
  • Rosa Fernández
  • Céline Fournier
  • Matilde Fuentes
  • Peter Galle
  • Edgar García
  • Montserrat García-Retortillo
  • Esther Garrido
  • Pere Ginès
  • Rosa Gordillo Medina
  • Jordi Gratacós-Ginès
  • Isabel Graupera
  • Ivica Grgurevic
  • Indra Neil Guha
  • Eva Guix
  • Johanne Kragh Hansen
  • Rebecca Harris
  • Elena Hernández Boluda
  • Rosario Hernández-Ibañez
  • Jordi Hoyo
  • Arfan Ikram
  • Simone Incicco
  • Mads Israelsen
  • Marta Juan
  • Adrià Juanola
  • Ralf Kaiser
  • Patrick S Kamath
  • Tom H Karlsen
  • Maria Kjærgaard
  • Marko Korenjak
  • Aleksander Krag
  • Marcin Krawczyk
  • Philippe Laboulaye
  • Irina Lambert
  • Frank Lammert
  • Simon Langkjær Sørensen
  • Cristina Laserna-Jiménez
  • Sonia Lazaro Pi
  • Elsa Ledain
  • Vincent Levy
  • Katrine Prier Lindvig
  • Anne Llorca
  • Vanessa Londoño
  • Guirec Loyer
  • Ann T. Ma
  • Anita Madir
  • Michael Manns
  • Denise Marshall
  • M. Lluïsa Martí
  • Sara Martínez
  • Ricard Martínez Sala
  • Roser Masa-Font
  • Jane Møller Jensen
  • Rosa M Morillas
  • Laura Muñoz
  • Ruth Nadal
  • Laura Napoleone
  • JM Navarrete
  • Phillip N Newsome
  • Vibeke Nielsen
  • Martina Pérez
  • Juan Manuel Pericás-Pulido
  • Salvatore Piano
  • Judit Pich
  • Elisa Pose
  • Judit Presas Escobet
  • Matthias Reichert
  • Carlota Riba
  • Dominique Roulot
  • Ana Belén Rubio
  • Maria Sánchez-Morata
  • Jörn Schattenberg
  • Miquel Serra-Burriel
  • Feliu Serra-Burriel
  • Louise Skovborg Just
  • Milan Sonneveld
  • Anna Soria
  • Christiane Stern
  • Patricia Such
  • Maja Thiele
  • Marta Tonon
  • Pere Toran
  • Antoni Torrejón
  • Emmanuel A Tsochatzis
  • Laurens van Kleef
  • Paulien van Wijngaarden
  • Vanessa Velázquez
  • Ana Viu
  • Susanne Nicole Weber
  • Tracey Wildsmith
https://doi.org/10.1016/S0140-6736(23)01174-1
The Lancet2023-08-01
An integrated view of anti-inflammatory and antifibrotic targets for the treatment of NASH
DOI: 10.1016/j.jhep.2023.03.038
Authors:
  • Frank Tacke
  • Tobias Puengel
  • Rohit Loomba
  • Scott L. Friedman
https://doi.org/10.1016/j.jhep.2023.03.038
Journal of Hepatology2023-08-01
Apical bulkheads accumulate as adaptive response to impaired bile flow in liver disease.
Hepatocytes form bile canaliculi that dynamically respond to the signalling activity of bile acids and bile flow. Little is known about their responses to intraluminal pressure. During embryonic development, hepatocytes assemble apical bulkheads that increase the canalicular resistance to intraluminal pressure. Here, we investigate whether they also protect bile canaliculi against elevated pressure upon impaired bile flow in adult liver. Apical bulkheads accumulate upon bile flow obstruction in mouse models and patients with primary sclerosing cholangitis (PSC). Their loss under these conditions leads to abnormally dilated canaliculi, resembling liver cell rosettes described in other hepatic diseases. 3D reconstruction reveals that these structures are sections of cysts and tubes formed by hepatocytes. Mathematical modelling establishes that they positively correlate with canalicular pressure and occur in early PSC stages. Using primary hepatocytes and 3D organoids, we demonstrate that excessive canalicular pressure causes the loss of apical bulkheads and formation of rosettes. Our results suggest that apical bulkheads are a protective mechanism of hepatocytes against impaired bile flow, highlighting the role of canalicular pressure in liver diseases.
Authors:
  • C. Mayer
  • S. Nehring
  • M. Kucken
  • U. Repnik
  • S. Seifert
  • A. Sljukic
  • J. Delpierre
  • H. Morales-Navarrete
  • S. Hinz
  • M. Brosch
  • B. Chung
  • T. Karlsen
  • M. Huch
  • Y. Kalaidzidis
  • L. Brusch
  • J. Hampe
  • C. Schafmayer
  • M. Zerial
https://www.ncbi.nlm.nih.gov/pubmed/37522754
EMBO Rep2023-07-31
To TIPS or Not to TIPS in High Risk of Variceal Rebleeding and Acute-on-Chronic Liver Failure.
Variceal bleeding is a consequence of severe portal hypertension in patients with liver cirrhosis. Although the rate of bleeding has decreased over time, variceal bleeding in the presence of acute-on-chronic liver failure (ACLF) carries a high risk of treatment failure and short-term mortality. Treatment and/or removal of precipitating events (mainly bacterial infection and alcoholic hepatitis) and decrease of portal pressure may improve outcome of patients with acute decompensation or ACLF. Transjugular intrahepatic portosystemic shunts (TIPSs), especially in the preemptive situation, have been found to efficiently control bleeding, prevent rebleeding, and reduce short-term mortality. Therefore, TIPS placement should be considered as an option in the management of ACLF patients with variceal bleeding.
Authors:
  • W. Gu
  • M. Kimmann
  • W. Laleman
  • M. Praktiknjo
  • J. Trebicka
https://www.ncbi.nlm.nih.gov/pubmed/37286178
Semin Liver Dis2023-07-17
Endoscopic Advances in Hepatology.
Endoscopy is and remains an indispensable tool in diagnosing and managing liver disease and its complications. Due to the progress in advanced endoscopy, endoscopy has become an alternative route for many surgical, percutaneous, and angiographic interventions, not only as a backup tool when conventional interventions fail but increasingly as a first-line choice. The term endo-hepatology refers to the integration of advanced endoscopy in the practice of hepatology. Endoscopy is key in the diagnosis and management of esophageal and gastric varices, portal hypertensive gastropathy, and gastric antral vascular ectasia. Endoscopic ultrasound (EUS) can be used for the evaluation of the liver parenchyma, liver lesions, and surrounding tissues and vessels, including targeted biopsy and complemented with new software functions. Moreover, EUS can guide portal pressure gradient measurement, and assess and help manage complications of portal hypertension. It is crucial that each present-day hepatologist is aware of the (rapidly increasing) full spectrum of diagnostic and therapeutic tools that exist within this field. In this comprehensive review, we would like to discuss the current endo-hepatology spectrum, as well as future directions for endoscopy in hepatology.
Authors:
  • E. Vanderschueren
  • J. Trebicka
  • W. Laleman
https://www.ncbi.nlm.nih.gov/pubmed/37192654
Semin Liver Dis2023-07-17
Basal MET Phosphorylation is an Indicator of Hepatocyte Dysregulation in Liver Disease
Chronic liver diseases are worldwide on the rise. Due to the rapidly increasing incidence, in particular in Western countries, Non-alcoholic fatty liver disease (NAFLD) is gaining importance as the disease can develop into hepatocellular carcinoma. Lipid accumulation in hepatocytes has been identified as the characteristic structural change in NAFLD development, but molecular mechanisms responsible for disease progression remained unresolved. Here, we uncover in primary hepatocytes from a preclinical model fed with a Western diet (WD) a strong downregulation of the PI3K-AKT pathway and an upregulation of the MAPK pathway. Dynamic pathway modeling of hepatocyte growth factor (HGF) signal transduction combined with global proteomics identifies that an elevated basal MET phosphorylation rate is the main driver of altered signaling leading to increased proliferation of WD-hepatocytes. Model-adaptation to patient-derived hepatocytes reveal patient-specific variability in basal MET phosphorylation, which correlates with patient outcome after liver surgery. Thus, dysregulated basal MET phosphorylation could be an indicator for the health status of the liver and thereby inform on the risk of a patient to suffer from liver failure after surgery.
DOI: 10.1101/2023.07.04.547655
Authors:
  • Sebastian Burbano de Lara
  • Svenja Kemmer
  • Ina Biermayer
  • Svenja Feiler
  • Artyom Vlasov
  • Lorenza A. D'Alessandro
  • Barbara Helm
  • Yannik Dieter
  • Ahmed Ghallab
  • Jan G Hengstler
  • Katrin Hoffmann
  • Marcel Schilling
  • Jens Timmer
  • Ursula Klingmueller
https://doi.org/10.1101/2023.07.04.547655
2023-07-04
Hepatic inflammatory responses in liver fibrosis.
Chronic liver diseases such as nonalcoholic fatty liver disease (NAFLD) or viral hepatitis are characterized by persistent inflammation and subsequent liver fibrosis. Liver fibrosis critically determines long-term morbidity (for example, cirrhosis or liver cancer) and mortality in NAFLD and nonalcoholic steatohepatitis (NASH). Inflammation represents the concerted response of various hepatic cell types to hepatocellular death and inflammatory signals, which are related to intrahepatic injury pathways or extrahepatic mediators from the gut-liver axis and the circulation. Single-cell technologies have revealed the heterogeneity of immune cell activation concerning disease states and the spatial organization within the liver, including resident and recruited macrophages, neutrophils as mediators of tissue repair, auto-aggressive features of T cells as well as various innate lymphoid cell and unconventional T cell populations. Inflammatory responses drive the activation of hepatic stellate cells (HSCs), and HSC subsets, in turn, modulate immune mechanisms via chemokines and cytokines or transdifferentiate into matrix-producing myofibroblasts. Current advances in understanding the pathogenesis of inflammation and fibrosis in the liver, mainly focused on NAFLD or NASH owing to the high unmet medical need, have led to the identification of several therapeutic targets. In this Review, we summarize the inflammatory mediators and cells in the diseased liver, fibrogenic pathways and their therapeutic implications.
Authors:
  • L. Hammerich
  • F. Tacke
https://www.ncbi.nlm.nih.gov/pubmed/37400694
Nat Rev Gastroenterol Hepatol2023-07-03
Tolerance of repeated toxic injuries of murine livers is associated with steatosis and inflammation
Abstract The human liver has a remarkable capacity to regenerate and thus compensate over decades for fibrosis caused by toxic chemicals, drugs, alcohol, or malnutrition. To date, no protective mechanisms have been identified that help the liver tolerate these repeated injuries. In this study, we revealed dysregulation of lipid metabolism and mild inflammation as protective mechanisms by studying longitudinal multi-omic measurements of liver fibrosis induced by repeated CCl 4 injections in mice ( n  = 45). Based on comprehensive proteomics, transcriptomics, blood- and tissue-level profiling, we uncovered three phases of early disease development—initiation, progression, and tolerance. Using novel multi-omic network analysis, we identified multi-level mechanisms that are significantly dysregulated in the injury-tolerant response. Public data analysis shows that these profiles are altered in human liver diseases, including fibrosis and early cirrhosis stages. Our findings mark the beginning of the tolerance phase as the critical switching point in liver response to repetitive toxic doses. After fostering extracellular matrix accumulation as an acute response, we observe a deposition of tiny lipid droplets in hepatocytes only in the Tolerant phase. Our comprehensive study shows that lipid metabolism and mild inflammation may serve as biomarkers and are putative functional requirements to resist further disease progression.
DOI: 10.1038/s41419-023-05855-4
Authors:
  • Seddik Hammad
  • Christoph Ogris
  • Amnah Othman
  • Pia Erdoesi
  • Wolfgang Schmidt-Heck
  • Ina Biermayer
  • Barbara Helm
  • Yan Gao
  • Weronika Piorońska
  • Christian H. Holland
  • Lorenza A. D’Alessandro
  • Carolina de la Torre
  • Carsten Sticht
  • Sherin Al Aoua
  • Fabian J. Theis
  • Heike Bantel
  • Matthias P. Ebert
  • Ursula Klingmüller
  • Jan G. Hengstler
  • Steven Dooley
  • Nikola S. Mueller
https://doi.org/10.1038/s41419-023-05855-4
Cell Death & Disease2023-07-01
Sublethal necroptosis signaling promotes inflammation and liver cancer
DOI: 10.1016/j.immuni.2023.05.017
Authors:
  • Mihael Vucur
  • Ahmed Ghallab
  • Anne T. Schneider
  • Arlind Adili
  • Mingbo Cheng
  • Mirco Castoldi
  • Michael T. Singer
  • Veronika Büttner
  • Leonie S. Keysberg
  • Lena Küsgens
  • Marlene Kohlhepp
  • Boris Görg
  • Suchira Gallage
  • Jose Efren Barragan Avila
  • Kristian Unger
  • Claus Kordes
  • Anne-Laure Leblond
  • Wiebke Albrecht
  • Sven H. Loosen
  • Carolin Lohr
  • Markus S. Jördens
  • Anne Babler
  • Sikander Hayat
  • David Schumacher
  • Maria T. Koenen
  • Olivier Govaere
  • Mark V. Boekschoten
  • Simone Jörs
  • Carlos Villacorta-Martin
  • Vincenzo Mazzaferro
  • Josep M. Llovet
  • Ralf Weiskirchen
  • Jakob N. Kather
  • Patrick Starlinger
  • Michael Trauner
  • Mark Luedde
  • Lara R. Heij
  • Ulf P. Neumann
  • Verena Keitel
  • Johannes G. Bode
  • Rebekka K. Schneider
  • Frank Tacke
  • Bodo Levkau
  • Twan Lammers
  • Georg Fluegen
  • Theodore Alexandrov
  • Amy L. Collins
  • Glyn Nelson
  • Fiona Oakley
  • Derek A. Mann
  • Christoph Roderburg
  • Thomas Longerich
  • Achim Weber
  • Augusto Villanueva
  • Andre L. Samson
  • James M. Murphy
  • Rafael Kramann
  • Fabian Geisler
  • Ivan G. Costa
  • Jan G. Hengstler
  • Mathias Heikenwalder
  • Tom Luedde
https://doi.org/10.1016/j.immuni.2023.05.017
Immunity2023-07-01
Gene regulatory network inference in the era of single-cell multi-omics.
The interplay between chromatin, transcription factors and genes generates complex regulatory circuits that can be represented as gene regulatory networks (GRNs). The study of GRNs is useful to understand how cellular identity is established, maintained and disrupted in disease. GRNs can be inferred from experimental data - historically, bulk omics data - and/or from the literature. The advent of single-cell multi-omics technologies has led to the development of novel computational methods that leverage genomic, transcriptomic and chromatin accessibility information to infer GRNs at an unprecedented resolution. Here, we review the key principles of inferring GRNs that encompass transcription factor-gene interactions from transcriptomics and chromatin accessibility data. We focus on the comparison and classification of methods that use single-cell multimodal data. We highlight challenges in GRN inference, in particular with respect to benchmarking, and potential further developments using additional data modalities.
Authors:
  • P. Badia-I-Mompel
  • L. Wessels
  • S. Muller-Dott
  • R. Trimbour
  • R. O. Ramirez Flores
  • R. Argelaguet
  • J. Saez-Rodriguez
https://www.ncbi.nlm.nih.gov/pubmed/37365273
Nat Rev Genet2023-06-26
Acute-On-Chronic Liver Failure: Current Interventional Treatment Options and Future Challenges.
Acute-on-chronic liver failure (ACLF) is a frequent complication in patients with liver cirrhosis that has high short-term mortality. It is characterized by acute decompensation (AD) of liver cirrhosis, intra- and extrahepatic organ failure, and severe systemic inflammation (SI). In the recent past, several studies have investigated the management of this group of patients. Identification and treatment of precipitants of decompensation and ACLF play an important role, and management of the respective intra- and extrahepatic organ failures is essential. However, no specific treatment for ACLF has been established to date, and the only curative treatment option currently available for these patients is liver transplantation (LT). It has been shown that ACLF patients are at severe risk of waitlist mortality, and post-LT survival rates are high, making ACLF patients suitable candidates for LT. However, only a limited number of patients are eligible for LT due to related contraindications such as uncontrolled infections. In this case, bridging strategies (e.g., extracorporeal organ support systems) are required. Further therapeutic approaches have recently been developed and evaluated. Thus, this review focuses on current management and potential future treatment options.
Authors:
  • M. Kimmann
  • J. Trebicka
https://www.ncbi.nlm.nih.gov/pubmed/37511665
J Pers Med2023-06-26
Efruxifermin, an investigational treatment for fibrotic or cirrhotic nonalcoholic steatohepatitis (NASH)
DOI: 10.1080/13543784.2023.2230115
Authors:
  • Tobias Puengel
  • Frank Tacke
https://doi.org/10.1080/13543784.2023.2230115
Expert Opinion on Investigational Drugs2023-06-03
Synthesis and In Vitro Biological Evaluation of p-Carborane-Based Di-tert-butylphenol Analogs
Targeting inflammatory mediators and related signaling pathways may offer a rational strategy for the treatment of cancer. The incorporation of metabolically stable, sterically demanding, and hydrophobic carboranes in dual cycloxygenase-2 (COX-2)/5-lipoxygenase (5-LO) inhibitors that are key enzymes in the biosynthesis of eicosanoids is a promising approach. The di-tert-butylphenol derivatives R-830, S-2474, KME-4, and E-5110 represent potent dual COX-2/5-LO inhibitors. The incorporation of p-carborane and further substitution of the p-position resulted in four carborane-based di-tert-butylphenol analogs that showed no or weak COX inhibition but high 5-LO inhibitory activities in vitro. Cell viability studies on five human cancer cell lines revealed that the p-carborane analogs R-830-Cb, S-2474-Cb, KME-4-Cb, and E-5110-Cb exhibited lower anticancer activity compared to the related di-tert-butylphenols. Interestingly, R-830-Cb did not affect the viability of primary cells and suppressed HCT116 cell proliferation more potently than its carbon-based R-830 counterpart. Considering all the advantages of boron cluster incorporation for enhancement of drug biostability, selectivity, and availability of drugs, R-830-Cb can be tested in further mechanistic and in vivo studies.
DOI: 10.3390/molecules28114547
Authors:
  • Sebastian Braun
  • Sanja Jelača
  • Markus Laube
  • Sven George
  • Bettina Hofmann
  • Peter Lönnecke
  • Dieter Steinhilber
  • Jens Pietzsch
  • Sanja Mijatović
  • Danijela Maksimović-Ivanić
  • Evamarie Hey-Hawkins
https://doi.org/10.3390/molecules28114547
Molecules2023-06-01
Carborane‐Based Tebufelone Analogs and Their Biological Evaluation In Vitro
The presence of inflammatory mediators in the tumor microenvironment, such as cytokines, growth factors or eicosanoids, indicate cancer-related inflammatory processes. Targeting these inflammatory mediators and related signal pathways may offer a rational strategy for the treatment of cancer. This study focuses on the incorporation of metabolically stable, sterically demanding, and hydrophobic dicarba-closo-dodecaboranes (carboranes) into dual cyclooxygenase-2 (COX-2)/5-lipoxygenase (5-LO) inhibitors that are key enzymes in the biosynthesis of eicosanoids. The di-tert-butylphenol derivative tebufelone represents a selective dual COX-2/5-LO inhibitor. The incorporation of meta- or para-carborane into the tebufelone scaffold resulted in eight carborane-based tebufelone analogs that show no COX inhibition but 5-LO inhibitory activity in vitro. Cell viability studies on HT29 colon adenocarcinoma cells revealed that the observed antiproliferative effect of the para-carborane analogs of tebufelone is enhanced by structural modifications that include chain elongation in combination with introduction of a methylene spacer resulting in higher anticancer activity compared to tebufelone. Hence, this strategy proved to be a promising approach to design potent 5-LO inhibitors with potential application as cytostatic agents.
DOI: 10.1002/cmdc.202300206
Authors:
  • Sebastian Braun
  • Svetlana Paskaš
  • Markus Laube
  • Sven George
  • Bettina Hofmann
  • Peter Lönnecke
  • Dieter Steinhilber
  • Jens Pietzsch
  • Sanja Mijatović
  • Danijela Maksimović‐Ivanić
  • Evamarie Hey‐Hawkins
https://doi.org/10.1002/cmdc.202300206
ChemMedChem2023-05-24
Isonimesulide and Its Carborane Analogues as Isoform‐Selective COX Inhibitors and Antitumor Agents
Nonsteroidal anti-inflammatory drugs (NSAIDs) are the most widely used therapeutics against pain, fever, and inflammation; additionally, antitumor properties are reported. NSAIDs reduce the synthesis of prostaglandins by inhibiting the cyclooxygenase (COX) isoforms COX-1 and COX-2. As nonselective inhibition is associated with off-target effects, strategies to achieve selectivity for the clinically preferred isoform COX-2 are of high interest. The modification of NSAIDs using carborane clusters as phenyl mimetics is reported to alter the selectivity profile through size exclusion. Inspired by these findings, isonimesulide and its carborane derivatives are prepared. The biological screening shows that the carborane containing compounds exhibit a stronger antitumor potential compared to nimesulide and isonimesulide. Furthermore, the replacement of the phenyl ring of isonimesulide with a carborane moiety resulted in a shift of the COX activity from nonactive to COX-active compounds.
DOI: 10.1002/adtp.202300117
Authors:
  • Liridona Useini
  • Teodora Komazec
  • Markus Laube
  • Peter Lönnecke
  • Jonas Schädlich
  • Sanja Mijatović
  • Danijela Maksimović‐Ivanić
  • Jens Pietzsch
  • Evamarie Hey‐Hawkins
https://doi.org/10.1002/adtp.202300117
Advanced Therapeutics2023-05-24
Cholangiokines: undervalued modulators in the hepatic microenvironment
The biliary epithelial cells, also known as cholangiocytes, line the intra- and extrahepatic bile ducts, forming a barrier between intra- and extra-ductal environments. Cholangiocytes are mostly known to modulate bile composition and transportation. In hepatobiliary diseases, bile duct injury leads to drastic alterations in cholangiocyte phenotypes and their release of soluble mediators, which can vary depending on the original insult and cellular states (quiescence, senescence, or proliferation). The cholangiocyte-secreted cytokines (also termed cholangiokines) drive ductular cell proliferation, portal inflammation and fibrosis, and carcinogenesis. Hence, despite the previous consensus that cholangiocytes are bystanders in liver diseases, their diverse secretome plays critical roles in modulating the intrahepatic microenvironment. This review summarizes recent insights into the cholangiokines under both physiological and pathological conditions, especially as they occur during liver injury-regeneration, inflammation, fibrosis and malignant transformation processes.
DOI: 10.3389/fimmu.2023.1192840
Authors:
  • Xiurong Cai
  • Frank Tacke
  • Adrien Guillot
  • Hanyang Liu
https://doi.org/10.3389/fimmu.2023.1192840
Frontiers in Immunology2023-05-16
The rationale and study design of two phase II trials examining the effects of BI 685,509, a soluble guanylyl cyclase activator, on clinically significant portal hypertension in patients with compensated cirrhosis.
BACKGROUND: Clinically significant portal hypertension (CSPH) drives cirrhosis-related complications (i.e. hepatic decompensation). Impaired nitric oxide (NO) bioavailability promotes sinusoidal vasoconstriction, which is the initial pathomechanism of CSPH development. Activation of soluble guanylyl cyclase (sGC), a key downstream effector of NO, facilitates sinusoidal vasodilation, which in turn may improve CSPH. Two phase II studies are being conducted to assess the efficacy of the NO-independent sGC activator BI 685,509 in patients with CSPH due to various cirrhosis aetiologies. METHODS: The 1366.0021 trial (NCT05161481) is a randomised, placebo-controlled, exploratory study that will assess BI 685,509 (moderate or high dose) for 24 weeks in patients with CSPH due to alcohol-related liver disease. The 1366.0029 trial (NCT05282121) is a randomised, open-label, parallel-group, exploratory study that will assess BI 685,509 (high dose) alone in patients with hepatitis B or C virus infection or non-alcoholic steatohepatitis (NASH) and in combination with 10 mg empagliflozin in patients with NASH and type 2 diabetes mellitus for 8 weeks. The 1366.0021 trial will enrol 105 patients, and the 1366.0029 trial will enrol 80 patients. In both studies, the primary endpoint is the change from baseline in hepatic venous pressure gradient (HVPG) until the end of treatment (24 or 8 weeks, respectively). Secondary endpoints include the proportion of patients with an HVPG reduction of > 10% from baseline, the development of decompensation events and the change from baseline in HVPG after 8 weeks in the 1366.0021 trial. In addition, the trials will assess changes in liver and spleen stiffness by transient elastography, changes in hepatic and renal function and the tolerability of BI 685,509. DISCUSSION: These trials will enable the assessment of the short-term (8 weeks) and longer-term (24 weeks) effects and safety of sGC activation by BI 685,509 on CSPH due to various cirrhosis aetiologies. The trials will use central readings of the diagnostic gold standard HVPG for the primary endpoint, as well as changes in established non-invasive biomarkers, such as liver and spleen stiffness. Ultimately, these trials will provide key information for developing future phase III trials. TRIAL REGISTRATION: 1366.0021: EudraCT no. 2021-001,285-38; ClinicalTrials.gov NCT05161481. Registered on 17 December 2021, https://www. CLINICALTRIALS: gov/ct2/show/NCT05161481 . 1366.0029: EudraCT no. 2021-005,171-40; ClinicalTrials.gov NCT05282121. Registered on 16 March 2022, https://www. CLINICALTRIALS: gov/ct2/show/NCT05282121 .
Authors:
  • T. Reiberger
  • A. Berzigotti
  • J. Trebicka
  • J. Ertle
  • I. Gashaw
  • R. Swallow
  • A. Tomisser
https://www.ncbi.nlm.nih.gov/pubmed/37095557
Trials2023-04-24
Hepatocyte apical bulkheads provide a mechanical means to oppose bile pressure
Hepatocytes grow their apical surfaces anisotropically to generate a 3D network of bile canaliculi (BC). BC elongation is ensured by apical bulkheads, membrane extensions that traverse the lumen and connect juxtaposed hepatocytes. We hypothesize that apical bulkheads are mechanical elements that shape the BC lumen in liver development but also counteract elevated biliary pressure. Here, by resolving their structure using STED microscopy, we found that they are sealed by tight junction loops, connected by adherens junctions, and contain contractile actomyosin, characteristics of mechanical function. Apical bulkheads persist at high pressure upon microinjection of fluid into the BC lumen, and laser ablation demonstrated that they are under tension. A mechanical model based on ablation results revealed that apical bulkheads double the pressure BC can hold. Apical bulkhead frequency anticorrelates with BC connectivity during mouse liver development, consistent with predicted changes in biliary pressure. Our findings demonstrate that apical bulkheads are load-bearing mechanical elements that could protect the BC network against elevated pressure.
DOI: 10.1083/jcb.202208002
Authors:
  • Maarten P. Bebelman
  • Matthew J. Bovyn
  • Carlotta M. Mayer
  • Julien Delpierre
  • Ronald Naumann
  • Nuno P. Martins
  • Alf Honigmann
  • Yannis Kalaidzidis
  • Pierre A. Haas
  • Marino Zerial
https://doi.org/10.1083/jcb.202208002
Journal of Cell Biology2023-04-03
Shell engineering in soft alginate‐based capsules for culturing liver spheroids
Functional interaction between cancer cells and the surrounding microenvironment is still not sufficiently understood, which motivates the tremendous interest for the development of numerous in vitro tumor models. Diverse parameters, for example, transport of nutrients and metabolites, availability of space in the confinement, etc. make an impact on the size, shape, and metabolism of the tumoroids. We demonstrate the fluidics-based low-cost methodology to reproducibly generate the alginate and alginate-chitosan microcapsules and apply it to grow human hepatoma (HepG2) spheroids of different dimensions and geometries. Focusing specifically on the composition and thickness of the hydrogel shell, permeability of the microcapsules was selectively tuned. The diffusion of the selected benchmark molecules through the shell has been systematically investigated using both, experiments and simulations, which is essential to ensure efficient mass transfer and/or filtering of the biochemical species. Metabolic activity of spheroids in microcapsules was confirmed by tracking the turnover of testosterone to androstenedione with chromatography studies in a metabolic assay. Depending on available space, phenotypically different 3D cell assemblies have been observed inside the capsules, varying in the tightness of cell aggregations and their shapes. Conclusively, we believe that our system with the facile tuning of the shell thickness and permeability, represents a promising platform for studying the formation of cancer spheroids and their functional interaction with the surrounding microenvironment.
DOI: 10.1002/biot.202200365
Authors:
  • Xuan Peng
  • Željko Janićijević
  • Sandy Lemm
  • Markus Laube
  • Jens Pietzsch
  • Michael Bachmann
  • Larysa Baraban
https://doi.org/10.1002/biot.202200365
Biotechnology Journal2023-03-27
Serum CXCL5 Detects Early Hepatocellular Carcinoma and Indicates Tumor Progression.
Chemokines or chemotactic cytokines play a pivotal role in the immune pathogenesis of liver cirrhosis and hepatocellular carcinoma (HCC). Nevertheless, comprehensive cytokine profiling data across different etiologies of liver diseases are lacking. Chemokines might serve as diagnostic and prognostic biomarkers. In our study, we analyzed serum concentrations of 12 inflammation-related chemokines in a cohort of patients (n = 222) with cirrhosis of different etiologies and/or HCC. We compared 97 patients with cirrhosis and treatment-naive HCC to the chemokine profile of 125 patients with cirrhosis but confirmed absence of HCC. Nine out of twelve chemokines were significantly elevated in sera of cirrhotic patients with HCC compared to HCC-free cirrhosis controls (CCL2, CCL11, CCL17, CCL20, CXCL1, CXCL5, CXCL9, CXCL10, CXCL11). Among those, CXCL5, CXCL9, CXCL10, and CXCL11 were significantly elevated in patients with early HCC according to the Barcelona Clinic Liver Cancer (BCLC) stages 0/A compared to cirrhotic controls without HCC. In patients with HCC, CXCL5 serum levels were associated with tumor progression, and levels of CCL20 and CXCL8 with macrovascular invasion. Importantly, our study identified CXCL5, CXCL9, and CXCL10 as universal HCC markers, independent from underlying etiology of cirrhosis. In conclusion, regardless of the underlying liver disease, patients with cirrhosis share an HCC-specific chemokine profile. CXCL5 may serve as a diagnostic biomarker in cirrhotic patients for early HCC detection as well as for tumor progression.
Authors:
  • A. Laschtowitz
  • J. Lambrecht
  • T. Puengel
  • F. Tacke
  • R. Mohr
https://www.ncbi.nlm.nih.gov/pubmed/36982370
Int J Mol Sci2023-03-10
Solid-Phase Parallel Synthesis of Dual Histone Deacetylase-Cyclooxygenase Inhibitors
Multi-target drugs (MTDs) are emerging alternatives to combination therapies. Since both histone deacetylases (HDACs) and cyclooxygenase-2 (COX-2) are known to be overexpressed in several cancer types, we herein report the design, synthesis, and biological evaluation of a library of dual HDAC-COX inhibitors. The designed compounds were synthesized via an efficient parallel synthesis approach using preloaded solid-phase resins. Biological in vitro assays demonstrated that several of the synthesized compounds possess pronounced inhibitory activities against HDAC and COX isoforms. The membrane permeability and inhibition of cellular HDAC activity of selected compounds were confirmed by whole-cell HDAC inhibition assays and immunoblot experiments. The most promising dual inhibitors, C3 and C4, evoked antiproliferative effects in the low micromolar concentration range and caused a significant increase in apoptotic cells. In contrast to previous reports, the simultaneous inhibition of HDAC and COX activity by dual HDAC-COX inhibitors or combination treatments with vorinostat and celecoxib did not result in additive or synergistic anticancer activities.
DOI: 10.3390/molecules28031061
Authors:
  • Luisa M. Bachmann
  • Maria Hanl
  • Felix Feller
  • Laura Sinatra
  • Andrea Schöler
  • Jens Pietzsch
  • Markus Laube
  • Finn K. Hansen
https://doi.org/10.3390/molecules28031061
Molecules2023-02-01
Optimization of extracellular matrix for primary human hepatocyte cultures using mixed collagen-Matrigel matrices.
Loss of differentiation of primary human hepatocytes (PHHs) ex vivo is a known problem of in vitro liver models. Culture optimizations using collagen type I and Matrigel reduce the dedifferentiation process but are not able to prevent it. While neither of these extracellular matrices (ECMs) on their own correspond to the authentic hepatic ECM, a combination of them could more closely resemble the in vivo situation. Our study aimed to systematically analyze the influence of mixed matrices composed of collagen type I and Matrigel on the maintenance and reestablishment of hepatic functions. Therefore, PHHs were cultured on mixed collagen-Matrigel matrices in monolayer and sandwich cultures and viability, metabolic capacity, differentiation markers, cellular arrangement and the cells' ability to repolarize and form functional bile canaliculi were assessed by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), functional assays and immunofluorescence microscopy. Our results show that mixed matrices were superior to pure matrices in maintaining metabolic capacity and hepatic differentiation. In contrast, Matrigel supplementation can impair the development of a proper hepatocytic polarization. Our systematic study helps to compose an optimized ECM to maintain and reestablish hepatic differentiation on cellular and multicellular levels in human liver models.
Authors:
  • L. Seidemann
  • S. Prinz
  • J. C. Scherbel
  • C. Gotz
  • D. Seehofer
  • G. Damm
https://www.ncbi.nlm.nih.gov/pubmed/36660192
EXCLI J2023-01-20
Genetic variation in TERT modifies the risk of hepatocellular carcinoma in alcohol-related cirrhosis: results from a genome-wide case-control study
Objective Hepatocellular carcinoma (HCC) often develops in patients with alcohol-related cirrhosis at an annual risk of up to 2.5%. Some host genetic risk factors have been identified but do not account for the majority of the variance in occurrence. This study aimed to identify novel susceptibility loci for the development of HCC in people with alcohol related cirrhosis. Design Patients with alcohol-related cirrhosis and HCC (cases: n=1214) and controls without HCC (n=1866), recruited from Germany, Austria, Switzerland, Italy and the UK, were included in a two-stage genome-wide association study using a case–control design. A validation cohort of 1520 people misusing alcohol but with no evidence of liver disease was included to control for possible association effects with alcohol misuse. Genotyping was performed using the InfiniumGlobal Screening Array (V.24v2, Illumina) and the OmniExpress Array (V.24v1-0a, Illumina). Results Associations with variants rs738409 in PNPLA3 and rs58542926 in TM6SF2 previously associated with an increased risk of HCC in patients with alcohol-related cirrhosis were confirmed at genome-wide significance. A novel locus rs2242652(A) in TERT (telomerase reverse transcriptase) was also associated with a decreased risk of HCC, in the combined meta-analysis, at genome-wide significance (p=6.41×10 −9 , OR=0.61 (95% CI 0.52 to 0.70). This protective association remained significant after correction for sex, age, body mass index and type 2 diabetes (p=7.94×10 −5 , OR=0.63 (95% CI 0.50 to 0.79). Carriage of rs2242652(A) in TERT was associated with an increased leucocyte telomere length (p=2.12×10 −44 ). Conclusion This study identifies rs2242652 in TERT as a novel protective factor for HCC in patients with alcohol-related cirrhosis.
DOI: 10.1136/gutjnl-2022-327196
Authors:
  • Stephan Buch
  • Hamish Innes
  • Philipp Ludwig Lutz
  • Hans Dieter Nischalke
  • Jens U Marquardt
  • Janett Fischer
  • Karl Heinz Weiss
  • Jonas Rosendahl
  • Astrid Marot
  • Marcin Krawczyk
  • Markus Casper
  • Frank Lammert
  • Florian Eyer
  • Arndt Vogel
  • Silke Marhenke
  • Johann von Felden
  • Rohini Sharma
  • Stephen Rahul Atkinson
  • Andrew McQuillin
  • Jacob Nattermann
  • Clemens Schafmayer
  • Andre Franke
  • Christian Strassburg
  • Marcella Rietschel
  • Heidi Altmann
  • Stefan Sulk
  • Veera Raghavan Thangapandi
  • Mario Brosch
  • Carolin Lackner
  • Rudolf E Stauber
  • Ali Canbay
  • Alexander Link
  • Thomas Reiberger
  • Mattias Mandorfer
  • Georg Semmler
  • Bernhard Scheiner
  • Christian Datz
  • Stefano Romeo
  • Stefano Ginanni Corradini
  • William Lucien Irving
  • Joanne R Morling
  • Indra Neil Guha
  • Eleanor Barnes
  • M Azim Ansari
  • Jocelyn Quistrebert
  • Luca Valenti
  • Sascha A Müller
  • Marsha Yvonne Morgan
  • Jean-François Dufour
  • Jonel Trebicka
  • Thomas Berg
  • Pierre Deltenre
  • Sebastian Mueller
  • Jochen Hampe
  • Felix Stickel
https://doi.org/10.1136/gutjnl-2022-327196
Gut2023-01-05
The Diagnostic Approach towards Combined Hepatocellular-Cholangiocarcinoma—State of the Art and Future Perspectives
Combined hepatocellular-cholangiocarcinoma (cHCC-CCA) is a rare primary liver cancer which displays clinicopathologic features of both hepatocellular (HCC) and cholangiocellular carcinoma (CCA). The similarity to HCC and CCA makes the diagnostic workup particularly challenging. Alpha-fetoprotein (AFP) and carbohydrate antigen 19-9 (CA 19-9) are blood tumour markers related with HCC and CCA, respectively. They can be used as diagnostic markers in cHCC-CCA as well, albeit with low sensitivity. The imaging features of cHCC-CCA overlap with those of HCC and CCA, dependent on the predominant histopathological component. Using the Liver Imaging and Reporting Data System (LI-RADS), as many as half of cHCC-CCAs may be falsely categorised as HCC. This is especially relevant since the diagnosis of HCC may be made without histopathological confirmation in certain cases. Thus, in instances of diagnostic uncertainty (e.g., simultaneous radiological HCC and CCA features, elevation of CA 19-9 and AFP, HCC imaging features and elevated CA 19-9, and vice versa) multiple image-guided core needle biopsies should be performed and analysed by an experienced pathologist. Recent advances in the molecular characterisation of cHCC-CCA, innovative diagnostic approaches (e.g., liquid biopsies) and methods to analyse multiple data points (e.g., clinical, radiological, laboratory, molecular, histopathological features) in an all-encompassing way (e.g., by using artificial intelligence) might help to address some of the existing diagnostic challenges.
DOI: 10.3390/cancers15010301
Authors:
  • Johannes Eschrich
  • Zuzanna Kobus
  • Dominik Geisel
  • Sebastian Halskov
  • Florian Roßner
  • Christoph Roderburg
  • Raphael Mohr
  • Frank Tacke
https://doi.org/10.3390/cancers15010301
Cancers2023-01-01
Mapping the hepatic immune landscape identifies monocytic macrophages as key drivers of steatohepatitis and cholangiopathy progression
DOI: 10.1097/HEP.0000000000000270
Authors:
  • Adrien Guillot
  • Marc Winkler
  • Milessa Silva Afonso
  • Abhishek Aggarwal
  • David Lopez
  • Hilmar Berger
  • Marlene S. Kohlhepp
  • Hanyang Liu
  • Burcin Özdirik
  • Johannes Eschrich
  • Jing Ma
  • Moritz Peiseler
  • Felix Heymann
  • Swetha Pendem
  • Sangeetha Mahadevan
  • Bin Gao
  • Lauri Diehl
  • Ruchi Gupta
  • Frank Tacke
https://doi.org/10.1097/HEP.0000000000000270
Hepatology2023-01-01
Hepatic C-X-C chemokine receptor type 6–expressing innate lymphocytes limit detrimental myeloid hyperactivation in acute liver injury
DOI: 10.1097/HC9.0000000000000102
Authors:
  • Felix Heymann
  • Jana C. Mossanen
  • Moritz Peiseler
  • Patricia M. Niemietz
  • Bruna Araujo David
  • Oliver Krenkel
  • Anke Liepelt
  • Matheus Batista Carneiro
  • Marlene S. Kohlhepp
  • Paul Kubes
  • Frank Tacke
https://doi.org/10.1097/HC9.0000000000000102
Hepatology Communications2023-01-01
Digital twin demonstrates significance of biomechanical growth control in liver regeneration after partial hepatectomy
DOI: 10.1016/j.isci.2022.105714
Authors:
  • Stefan Hoehme
  • Seddik Hammad
  • Jan Boettger
  • Brigitte Begher-Tibbe
  • Petru Bucur
  • Eric Vibert
  • Rolf Gebhardt
  • Jan G. Hengstler
  • Dirk Drasdo
https://doi.org/10.1016/j.isci.2022.105714
iScience2023-01-01
In Vitro Models for the Study of Liver Biology and Diseases: Advances and Limitations
DOI: 10.1016/j.jcmgh.2022.11.008
Authors:
  • Savneet Kaur
  • Srivatsan Kidambi
  • Martí Ortega-Ribera
  • Le Thi Thanh Thuy
  • Natalia Nieto
  • Victoria C. Cogger
  • Wei-Fen Xie
  • Frank Tacke
  • Jordi Gracia-Sancho
https://doi.org/10.1016/j.jcmgh.2022.11.008
Cellular and Molecular Gastroenterology and Hepatology2023-01-01
Therapeutic modulation of the liver immune microenvironment
DOI: 10.1097/HEP.0000000000000386
Authors:
  • Herbert Tilg
  • Timon E. Adolph
  • Frank Tacke
https://doi.org/10.1097/HEP.0000000000000386
Hepatology2023-01-01
The Multifaceted Roles of Macrophages in NAFLD Pathogenesis
DOI: 10.1016/j.jcmgh.2023.03.002
Authors:
  • Joscha Vonderlin
  • Triantafyllos Chavakis
  • Michael Sieweke
  • Frank Tacke
https://doi.org/10.1016/j.jcmgh.2023.03.002
Cellular and Molecular Gastroenterology and Hepatology2023-01-01
Hepatocytes reprogram liver macrophages involving control of TGF-β activation, influencing liver regeneration and injury
Background: Macrophages play an important role in maintaining liver homeostasis and regeneration. However, it is not clear to what extent the different macrophage populations of the liver differ in terms of their activation state and which other liver cell populations may play a role in regulating the same. Methods: Reverse transcription PCR, flow cytometry, transcriptome, proteome, secretome, single cell analysis, and immunohistochemical methods were used to study changes in gene expression as well as the activation state of macrophages in vitro and in vivo under homeostatic conditions and after partial hepatectomy. Results: We show that F4/80+/CD11bhi/CD14hi macrophages of the liver are recruited in a C-C motif chemokine receptor (CCR2)–dependent manner and exhibit an activation state that differs substantially from that of the other liver macrophage populations, which can be distinguished on the basis of CD11b and CD14 expressions. Thereby, primary hepatocytes are capable of creating an environment in vitro that elicits the same specific activation state in bone marrow–derived macrophages as observed in F4/80+/CD11bhi/CD14hi liver macrophages in vivo. Subsequent analyses, including studies in mice with a myeloid cell–specific deletion of the TGF-β type II receptor, suggest that the availability of activated TGF-β and its downregulation by a hepatocyte-conditioned milieu are critical. Reduction of TGF-βRII-mediated signal transduction in myeloid cells leads to upregulation of IL-6, IL-10, and SIGLEC1 expression, a hallmark of the activation state of F4/80+/CD11bhi/CD14hi macrophages, and enhances liver regeneration. Conclusions: The availability of activated TGF-β determines the activation state of specific macrophage populations in the liver, and the observed rapid transient activation of TGF-β may represent an important regulatory mechanism in the early phase of liver regeneration in this context.
DOI: 10.1097/HC9.0000000000000208
Authors:
  • Stephanie D. Wolf
  • Christian Ehlting
  • Sophia Müller-Dott
  • Gereon Poschmann
  • Patrick Petzsch
  • Tobias Lautwein
  • Sai Wang
  • Barbara Helm
  • Marcel Schilling
  • Julio Saez-Rodriguez
  • Mihael Vucur
  • Kai Stühler
  • Karl Köhrer
  • Frank Tacke
  • Steven Dooley
  • Ursula Klingmüller
  • Tom Luedde
  • Johannes G. Bode
https://doi.org/10.1097/HC9.0000000000000208
Hepatology Communications2023-01-01
Guided interactive image segmentation using machine learning and color-based image set clustering.
MOTIVATION: Over the last decades, image processing and analysis have become one of the key technologies in systems biology and medicine. The quantification of anatomical structures and dynamic processes in living systems is essential for understanding the complex underlying mechanisms and allows, i.e. the construction of spatio-temporal models that illuminate the interplay between architecture and function. Recently, deep learning significantly improved the performance of traditional image analysis in cases where imaging techniques provide large amounts of data. However, if only a few images are available or qualified annotations are expensive to produce, the applicability of deep learning is still limited. RESULTS: We present a novel approach that combines machine learning-based interactive image segmentation using supervoxels with a clustering method for the automated identification of similarly colored images in large image sets which enables a guided reuse of interactively trained classifiers. Our approach solves the problem of deteriorated segmentation and quantification accuracy when reusing trained classifiers which is due to significant color variability prevalent and often unavoidable in biological and medical images. This increase in efficiency improves the suitability of interactive segmentation for larger image sets, enabling efficient quantification or the rapid generation of training data for deep learning with minimal effort. The presented methods are applicable for almost any image type and represent a useful tool for image analysis tasks in general. AVAILABILITY AND IMPLEMENTATION: The presented methods are implemented in our image processing software TiQuant which is freely available at tiquant.hoehme.com. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.
Authors:
  • A. Friebel
  • T. Johann
  • D. Drasdo
  • S. Hoehme
https://www.ncbi.nlm.nih.gov/pubmed/35976110
Bioinformatics2022-09-30
Erythropoietin-driven dynamic proteome adaptations during erythropoiesis prevent iron overload in the developing embryo.
Erythropoietin (Epo) ensures survival and proliferation of colony-forming unit erythroid (CFU-E) progenitor cells and their differentiation to hemoglobin-containing mature erythrocytes. A lack of Epo-induced responses causes embryonic lethality, but mechanisms regulating the dynamic communication of cellular alterations to the organismal level remain unresolved. By time-resolved transcriptomics and proteomics, we show that Epo induces in CFU-E cells a gradual transition from proliferation signature proteins to proteins indicative for differentiation, including heme-synthesis enzymes. In the absence of the Epo receptor (EpoR) in embryos, we observe a lack of hemoglobin in CFU-E cells and massive iron overload of the fetal liver pointing to a miscommunication between liver and placenta. A reduction of iron-sulfur cluster-containing proteins involved in oxidative phosphorylation in these embryos leads to a metabolic shift toward glycolysis. This link connecting erythropoiesis with the regulation of iron homeostasis and metabolic reprogramming suggests that balancing these interactions is crucial for protection from iron intoxication and for survival.
Authors:
  • S. Chakraborty
  • G. Andrieux
  • P. Kastl
  • L. Adlung
  • S. Altamura
  • M. E. Boehm
  • L. E. Schwarzmuller
  • Y. Abdullah
  • M. C. Wagner
  • B. Helm
  • H. J. Grone
  • W. D. Lehmann
  • M. Boerries
  • H. Busch
  • M. U. Muckenthaler
  • M. Schilling
  • U. Klingmuller
https://www.ncbi.nlm.nih.gov/pubmed/36130519
Cell Rep2022-09-20
Investigation of Radiotracer Metabolic Stability In Vitro with CYP-Overexpressing Hepatoma Cell Lines.
The characterization of novel radiotracers toward their metabolic stability is an essential part of their development. While in vitro methods such as liver microsome assays or ex vivo blood or tissue samples provide information on overall stability, little or no information is obtained on cytochrome P450 (CYP) enzyme and isoform-specific contribution to the metabolic fate of individual radiotracers. Herein, we investigated recently established CYP-overexpressing hepatoblastoma cell lines (HepG2) for their suitability to study the metabolic stability of radiotracers in general and to gain insight into CYP isoform specificity. Wildtype HepG2 and CYP1A2-, CYP2C19-, and CYP3A4-overexpressing HepG2 cells were incubated with radiotracers, and metabolic turnover was analyzed. The optimized protocol, covering cell seeding in 96-well plates and analysis of supernatant by radio thin-layer-chromatography for higher throughput, was transferred to the evaluation of three (18)F-labeled celecoxib-derived cyclooxygenase-2 inhibitors (coxibs). These investigations revealed time-dependent degradation of the intact radiotracers, as well as CYP isoform- and substrate-specific differences in their metabolic profiles. HepG2 CYP2C19 proved to be the cell line showing the highest metabolic turnover for each radiotracer studied here. Comparison with human and murine liver microsome assays showed good agreement with the human metabolite profile obtained by the HepG2 cell lines. Therefore, CYP-overexpressing HepG2 cells provide a good complement for assessing the metabolic stability of radiotracers and allow the analysis of the CYP isoform-specific contribution to the overall radiotracer metabolism.
Authors:
  • S. Lemm
  • S. Kohler
  • R. Wodtke
  • F. Jung
  • J. H. Kupper
  • J. Pietzsch
  • M. Laube
https://www.ncbi.nlm.nih.gov/pubmed/35954291
Cells2022-08-07
Deciphering signal transduction networks in the liver by mechanistic mathematical modelling
In health and disease, liver cells are continuously exposed to cytokines and growth factors. While individual signal transduction pathways induced by these factors were studied in great detail, the cellular responses induced by repeated or combined stimulations are complex and less understood. Growth factor receptors on the cell surface of hepatocytes were shown to be regulated by receptor interactions, receptor trafficking and feedback regulation. Here, we exemplify how mechanistic mathematical modelling based on quantitative data can be employed to disentangle these interactions at the molecular level. Crucial is the analysis at a mechanistic level based on quantitative longitudinal data within a mathematical framework. In such multi-layered information, step-wise mathematical modelling using submodules is of advantage, which is fostered by sharing of standardized experimental data and mathematical models. Integration of signal transduction with metabolic regulation in the liver and mechanistic links to translational approaches promise to provide predictive tools for biology and personalized medicine.
DOI: 10.1042/BCJ20210548
Authors:
  • Lorenza A. D’Alessandro
  • Ursula Klingmüller
  • Marcel Schilling
https://doi.org/10.1042/BCJ20210548
Biochemical Journal2022-06-30
Fibrogenic Pathways in Metabolic Dysfunction Associated Fatty Liver Disease (MAFLD).
The prevalence of nonalcoholic fatty liver disease (NAFLD), recently also re-defined as metabolic dysfunction associated fatty liver disease (MAFLD), is rapidly increasing, affecting ~25% of the world population. MALFD/NAFLD represents a spectrum of liver pathologies including the more benign hepatic steatosis and the more advanced non-alcoholic steatohepatitis (NASH). NASH is associated with enhanced risk for liver fibrosis and progression to cirrhosis and hepatocellular carcinoma. Hepatic stellate cells (HSC) activation underlies NASH-related fibrosis. Here, we discuss the profibrogenic pathways, which lead to HSC activation and fibrogenesis, with a particular focus on the intercellular hepatocyte-HSC and macrophage-HSC crosstalk.
Authors:
  • P. Subramanian
  • J. Hampe
  • F. Tacke
  • T. Chavakis
https://www.ncbi.nlm.nih.gov/pubmed/35805998
Int J Mol Sci2022-06-23
Diploid hepatocytes drive physiological liver renewal in adult humans
Physiological liver cell replacement is central to maintaining the organ’s high metabolic activity, although its characteristics are difficult to study in humans. Using retrospective radiocarbon (14C) birth dating of cells, we report that human hepatocytes show continuous and lifelong turnover, allowing the liver to remain a young organ (average age <3 years). Hepatocyte renewal is highly dependent on the ploidy level. Diploid hepatocytes show more than 7-fold higher annual birth rates than polyploid hepatocytes. These observations support the view that physiological liver cell renewal in humans is mainly dependent on diploid hepatocytes, whereas polyploid cells are compromised in their ability to divide. Moreover, cellular transitions between diploid and polyploid hepatocytes are limited under homeostatic conditions. With these findings, we present an integrated model of homeostatic liver cell generation in humans that provides fundamental insights into liver cell turnover dynamics.
DOI: 10.1016/j.cels.2022.05.001
Authors:
  • Paula Heinke
  • Fabian Rost
  • Julian Rode
  • Palina Trus
  • Irina Simonova
  • Enikő Lázár
  • Joshua Feddema
  • Thilo Welsch
  • Kanar Alkass
  • Mehran Salehpour
  • Andrea Zimmermann
  • Daniel Seehofer
  • Göran Possnert
  • Georg Damm
  • Henrik Druid
  • Lutz Brusch
  • Olaf Bergmann
https://doi.org/10.1016/j.cels.2022.05.001
Cell Systems2022-06-01
Hepatic Hedgehog Signaling Participates in the Crosstalk between Liver and Adipose Tissue in Mice by Regulating FGF21.
The Hedgehog signaling pathway regulates many processes during embryogenesis and the homeostasis of adult organs. Recent data suggest that central metabolic processes and signaling cascades in the liver are controlled by the Hedgehog pathway and that changes in hepatic Hedgehog activity also affect peripheral tissues, such as the reproductive organs in females. Here, we show that hepatocyte-specific deletion of the Hedgehog pathway is associated with the dramatic expansion of adipose tissue in mice, the overall phenotype of which does not correspond to the classical outcome of insulin resistance-associated diabetes type 2 obesity. Rather, we show that alterations in the Hedgehog signaling pathway in the liver lead to a metabolic phenotype that is resembling metabolically healthy obesity. Mechanistically, we identified an indirect influence on the hepatic secretion of the fibroblast growth factor 21, which is regulated by a series of signaling cascades that are directly transcriptionally linked to the activity of the Hedgehog transcription factor GLI1. The results of this study impressively show that the metabolic balance of the entire organism is maintained via the activity of morphogenic signaling pathways, such as the Hedgehog cascade. Obviously, several pathways are orchestrated to facilitate liver metabolic status to peripheral organs, such as adipose tissue.
Authors:
  • F. Ott
  • C. Korner
  • K. Werner
  • M. Gericke
  • I. Liebscher
  • D. Lobsien
  • S. Radrezza
  • A. Shevchenko
  • U. Hofmann
  • J. Kratzsch
  • R. Gebhardt
  • T. Berg
  • M. Matz-Soja
https://www.ncbi.nlm.nih.gov/pubmed/35626717
Cells2022-05-18
Hepatic Hedgehog Signaling Participates in the Crosstalk between Liver and Adipose Tissue in Mice by Regulating FGF21
The Hedgehog signaling pathway regulates many processes during embryogenesis and the homeostasis of adult organs. Recent data suggest that central metabolic processes and signaling cascades in the liver are controlled by the Hedgehog pathway and that changes in hepatic Hedgehog activity also affect peripheral tissues, such as the reproductive organs in females. Here, we show that hepatocyte-specific deletion of the Hedgehog pathway is associated with the dramatic expansion of adipose tissue in mice, the overall phenotype of which does not correspond to the classical outcome of insulin resistance-associated diabetes type 2 obesity. Rather, we show that alterations in the Hedgehog signaling pathway in the liver lead to a metabolic phenotype that is resembling metabolically healthy obesity. Mechanistically, we identified an indirect influence on the hepatic secretion of the fibroblast growth factor 21, which is regulated by a series of signaling cascades that are directly transcriptionally linked to the activity of the Hedgehog transcription factor GLI1. The results of this study impressively show that the metabolic balance of the entire organism is maintained via the activity of morphogenic signaling pathways, such as the Hedgehog cascade. Obviously, several pathways are orchestrated to facilitate liver metabolic status to peripheral organs, such as adipose tissue.
DOI: 10.3390/cells11101680
Authors:
  • Fritzi Ott
  • Christiane Körner
  • Kim Werner
  • Martin Gericke
  • Ines Liebscher
  • Donald Lobsien
  • Silvia Radrezza
  • Andrej Shevchenko
  • Ute Hofmann
  • Jürgen Kratzsch
  • Rolf Gebhardt
  • Thomas Berg
  • Madlen Matz-Soja
https://doi.org/10.3390/cells11101680
Cells2022-05-01
Transforming growth factor β latency: A mechanism of cytokine storage and signalling regulation in liver homeostasis and disease
DOI: 10.1016/j.jhepr.2021.100397
Authors:
  • Yujia Li
  • Weiguo Fan
  • Frederik Link
  • Sai Wang
  • Steven Dooley
https://doi.org/10.1016/j.jhepr.2021.100397
JHEP Reports2022-02-01
MSPypeline: a python package for streamlined data analysis of mass spectrometry-based proteomics
Abstract Summary Mass spectrometry-based proteomics is increasingly employed in biology and medicine. To generate reliable information from large datasets and ensure comparability of results, it is crucial to implement and standardize the quality control of the raw data, the data processing steps and the statistical analyses. MSPypeline provides a platform for importing MaxQuant output tables, generating quality control reports, data preprocessing including normalization and performing exploratory analyses by statistical inference plots. These standardized steps assess data quality, provide customizable figures and enable the identification of differentially expressed proteins to reach biologically relevant conclusions. Availability and implementation The source code is available under the MIT license at https://github.com/siheming/mspypeline with documentation at https://mspypeline.readthedocs.io. Benchmark mass spectrometry data are available on ProteomeXchange (PXD025792). Supplementary information Supplementary data are available at Bioinformatics Advances online.
DOI: 10.1093/bioadv/vbac004
Authors:
  • Simon Heming
  • Pauline Hansen
  • Artyom Vlasov
  • Florian Schwörer
  • Stephen Schaumann
  • Paulina Frolovaitė
  • Wolf-Dieter Lehmann
  • Jens Timmer
  • Marcel Schilling
  • Barbara Helm
  • Ursula Klingmüller
https://doi.org/10.1093/bioadv/vbac004
Bioinformatics Advances2022-01-01
The rs429358 Locus in Apolipoprotein E Is Associated With Hepatocellular Carcinoma in Patients With Cirrhosis
The host genetic background for hepatocellular carcinoma (HCC) is incompletely understood. We aimed to determine if four germline genetic polymorphisms, rs429358 in apolipoprotein E ( APOE ), rs2642438 in mitochondrial amidoxime reducing component 1 ( MARC1 ), rs2792751 in glycerol‐3‐phosphate acyltransferase ( GPAM ), and rs187429064 in transmembrane 6 superfamily member 2 ( TM6SF2 ), previously associated with progressive alcohol‐related and nonalcoholic fatty liver disease, are also associated with HCC. Four HCC case‐control data sets were constructed, including two mixed etiology data sets (UK Biobank and FinnGen); one hepatitis C virus (HCV) cohort (STOP‐HCV), and one alcohol‐related HCC cohort (Dresden HCC). The frequency of each variant was compared between HCC cases and cirrhosis controls (i.e., patients with cirrhosis without HCC). Population controls were also considered. Odds ratios (ORs) associations were calculated using logistic regression, adjusting for age, sex, and principal components of genetic ancestry. Fixed‐effect meta‐analysis was used to determine the pooled effect size across all data sets. Across four case‐control data sets, 2,070 HCC cases, 4,121 cirrhosis controls, and 525,779 population controls were included. The rs429358:C allele ( APOE ) was significantly less frequent in HCC cases versus cirrhosis controls (OR, 0.71; 95% confidence interval [CI], 0.61‐0.84; P  = 2.9 × 10 −5 ). Rs187429064:G ( TM6SF2 ) was significantly more common in HCC cases versus cirrhosis controls and exhibited the strongest effect size (OR, 2.03; 95% CI, 1.45‐2.86; P  = 3.1 × 10 −6 ). In contrast, rs2792751:T ( GPAM ) was not associated with HCC (OR, 1.01; 95% CI, 0.90‐1.13; P  = 0.89), whereas rs2642438:A ( MARC1 ) narrowly missed statistical significance (OR, 0.91; 95% CI, 0.84‐1.00; P  = 0.043). Conclusion: This study associates carriage of rs429358:C ( APOE ) with a reduced risk of HCC in patients with cirrhosis. Conversely, carriage of rs187429064:G in TM6SF2 is associated with an increased risk of HCC in patients with cirrhosis.
DOI: 10.1002/hep4.1886
Authors:
  • Hamish Innes
  • Hans Dieter Nischalke
  • Indra Neil Guha
  • Karl Heinz Weiss
  • Will Irving
  • Daniel Gotthardt
  • Eleanor Barnes
  • Janett Fischer
  • M. Azim Ansari
  • Jonas Rosendahl
  • Shang‐Kuan Lin
  • Astrid Marot
  • Vincent Pedergnana
  • Markus Casper
  • Jennifer Benselin
  • Frank Lammert
  • John McLauchlan
  • Philip L. Lutz
  • Victoria Hamill
  • Sebastian Mueller
  • Joanne R. Morling
  • Georg Semmler
  • Florian Eyer
  • Johann von Felden
  • Alexander Link
  • Arndt Vogel
  • Jens U. Marquardt
  • Stefan Sulk
  • Jonel Trebicka
  • Luca Valenti
  • Christian Datz
  • Thomas Reiberger
  • Clemens Schafmayer
  • Thomas Berg
  • Pierre Deltenre
  • Jochen Hampe
  • Felix Stickel
  • Stephan Buch
https://doi.org/10.1002/hep4.1886
Hepatology Communications2022-01-01