9 results on '"Iván M. Moya"'
Search Results
2. In Vivo Tracking and 3D Mapping of Cell Death in Regeneration and Cancer Using Trypan Blue
- Author
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Nicole Procel, Karen Camacho, Elisabeth Verboven, Isabel Baroja, Priscila A. Guerrero, Hanne Hillen, Carlos Estrella-García, Nicole Vizcaíno-Rodríguez, Leticia Sansores-Garcia, Ana Santamaría-Naranjo, Andrés Romero-Carvajal, Andrés Caicedo, Georg Halder, and Iván M. Moya
- Subjects
in vivo cell death tracking ,Trypan Blue labeling ,organ regeneration ,anticancer therapy assessment ,ischemia-reperfusion injury ,cholangiocarcinoma ,Cytology ,QH573-671 - Abstract
Tracking cell death in vivo can enable a better understanding of the biological mechanisms underlying tissue homeostasis and disease. Unfortunately, existing cell death labeling methods lack compatibility with in vivo applications or suffer from low sensitivity, poor tissue penetration, and limited temporal resolution. Here, we fluorescently labeled dead cells in vivo with Trypan Blue (TBlue) to detect single scattered dead cells or to generate whole-mount three-dimensional maps of large areas of necrotic tissue during organ regeneration. TBlue effectively marked different types of cell death, including necrosis induced by CCl4 intoxication in the liver, necrosis caused by ischemia-reperfusion in the skin, and apoptosis triggered by BAX overexpression in hepatocytes. Moreover, due to its short circulating lifespan in blood, TBlue labeling allowed in vivo “pulse and chase” tracking of two temporally spaced populations of dying hepatocytes in regenerating mouse livers. Additionally, upon treatment with cisplatin, TBlue labeled dead cancer cells in livers with cholangiocarcinoma and dead thymocytes due to chemotherapy-induced toxicity, showcasing its utility in assessing anticancer therapies in preclinical models. Thus, TBlue is a sensitive and selective cell death marker for in vivo applications, facilitating the understanding of the fundamental role of cell death in normal biological processes and its implications in disease.
- Published
- 2024
- Full Text
- View/download PDF
3. Comparison of the Opn-CreER and Ck19-CreER Drivers in Bile Ducts of Normal and Injured Mouse Livers
- Author
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Bram Lesaffer, Elisabeth Verboven, Leen Van Huffel, Iván M. Moya, Leo A. van Grunsven, Isabelle A. Leclercq, Frédéric P. Lemaigre, and Georg Halder
- Subjects
Cre ,cholangiocytes ,bile duct cells ,knockout ,mouse liver ,lineage tracing ,Opn ,Ck19 ,Cytology ,QH573-671 - Abstract
Inducible cyclization recombinase (Cre) transgenic mouse strains are powerful tools for cell lineage tracing and tissue-specific knockout experiments. However, low efficiency or leaky expression can be important pitfalls. Here, we compared the efficiency and specificity of two commonly used cholangiocyte-specific Cre drivers, the Opn-iCreERT2 and Ck19-CreERT drivers, using a tdTomato reporter strain. We found that Opn-iCreERT2 triggered recombination of the tdTomato reporter in 99.9% of all cholangiocytes while Ck19-CreERT only had 32% recombination efficiency after tamoxifen injection. In the absence of tamoxifen, recombination was also induced in 2% of cholangiocytes for the Opn-iCreERT2 driver and in 13% for the Ck19-CreERT driver. For both drivers, Cre recombination was highly specific for cholangiocytes since recombination was rare in other liver cell types. Toxic liver injury ectopically activated Opn-iCreERT2 but not Ck19-CreERT expression in hepatocytes. However, ectopic recombination in hepatocytes could be avoided by applying a three-day long wash-out period between tamoxifen treatment and toxin injection. Therefore, the Opn-iCreERT2 driver is best suited for the generation of mutant bile ducts, while the Ck19-CreERT driver has near absolute specificity for bile duct cells and is therefore favorable for lineage tracing experiments.
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- 2019
- Full Text
- View/download PDF
4. YAP and TAZ Heterogeneity in Primary Liver Cancer: An Analysis of Its Prognostic and Diagnostic Role
- Author
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Matthias Van Haele, Iván M. Moya, Ruçhan Karaman, Guy Rens, Janne Snoeck, Olivier Govaere, Frederik Nevens, Chris Verslype, Baki Topal, Diethard Monbaliu, Georg Halder, and Tania Roskams
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HCC ,cHCC-CCA ,CCA ,YAP ,TAZ ,Keratin 19 ,Hepatic progenitor cells ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Primary liver cancer comprises a diverse group of liver tumors. The heterogeneity of these tumors is seen as one of the obstacles to finding an effective therapy. The Hippo pathway, with its downstream transcriptional co-activator Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ), has a decisive role in the carcinogenesis of primary liver cancer. Therefore, we examined the expression pattern of YAP and TAZ in 141 patients with hepatocellular carcinoma keratin 19 positive (HCC K19+), hepatocellular carcinoma keratin 19 negative (HCC K19−), combined hepatocellular⁻cholangiocarcinoma carcinoma (cHCC-CCA), or cholangiocarcinoma (CCA). All cHCC-CCA and CCA patients showed high expression levels for YAP and TAZ, while only some patients of the HCC group were positive. Notably, we found that a histoscore of both markers is useful in the challenging diagnosis of cHCC-CCA. In addition, positivity for YAP and TAZ was observed in the hepatocellular and cholangiocellular components of cHCC-CCA, which suggests a single cell origin in cHCC-CCA. Within the K19− HCC group, our results demonstrate that the expression of YAP is a statistically significant predictor of poor prognosis when observed in the cytoplasm. Nuclear expression of TAZ is an even more specific and independent predictor of poor disease-free survival and overall survival of K19− HCC patients. Our results thus identify different levels of YAP/TAZ expression in various liver cancers that can be used for diagnostics.
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- 2019
- Full Text
- View/download PDF
5. Peritumoral activation of the Hippo pathway effectors YAP and TAZ suppresses liver cancer in mice
- Author
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Panagiotis Karras, Maxime de Waegeneer, Ruchan Karaman, Leticia Sansores-Garcia, Laura Van den Mooter, Inge Mannaerts, Randy L. Johnson, Hanne Hillen, Elisabeth Verboven, Matthias Van Haele, Weronika Kowalczyk, Iván M. Moya, Jun Xie, Tania Roskams, Georg Halder, Jean-Christophe Marine, Leen Van Huffel, Soheil Soheily, Lars Zender, Leo A. van Grunsven, Stefaan Verhulst, Stephanie Anais Castaldo, Ana Algueró-Nadal, Jelle Jacobs, Stein Aerts, Basic (bio-) Medical Sciences, Liver Cell Biology, and Translational Liver Cell Biology
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Carcinoma, Hepatocellular ,Cell Survival ,Cell ,Cell Cycle Proteins ,Protein Serine-Threonine Kinases ,Cholangiocarcinoma ,03 medical and health sciences ,Liver Neoplasms, Experimental ,0302 clinical medicine ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Hippo Signaling Pathway ,Melanoma ,Adaptor Proteins, Signal Transducing ,030304 developmental biology ,0303 health sciences ,Hippo signaling pathway ,Multidisciplinary ,Hyperactivation ,Chemistry ,Effector ,Liver Neoplasms ,Signal transducing adaptor protein ,YAP-Signaling Proteins ,medicine.disease ,Tumor Burden ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Cell culture ,Transcriptional Coactivator with PDZ-Binding Motif Proteins ,030220 oncology & carcinogenesis ,Hepatocytes ,Trans-Activators ,Cancer research ,Signal transduction ,Liver cancer ,Signal Transduction ,Transcription Factors - Abstract
Mixed signals at tumor margins The Hippo signaling pathway has been implicated in tumor growth, sparking interest in the pathway as a potential therapeutic target. In a study of liver cancer in genetically manipulated mice, Moya et al. discovered that the role of this pathway in tumorigenesis is more complex than previously appreciated. They confirmed that activation of the Hippo pathway within tumor cells drives tumor growth; however, they also found that activation of the pathway in adjacent healthy cells has the opposite effect, suppressing tumor growth. Whether tumor cells survive or are eliminated thus appears to depend on competing signals produced by the tumor and surrounding tissue. Science , this issue p. 1029
- Published
- 2019
6. Comparison of the Opn-CreER and Ck19-CreER Drivers in Bile Ducts of Normal and Injured Mouse Livers
- Author
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Leen Van Huffel, Georg Halder, Iván M. Moya, Isabelle Leclercq, Bram Lesaffer, Leo A. van Grunsven, Frédéric P. Lemaigre, Elisabeth Verboven, UCL - SSS/DDUV - Institut de Duve, UCL - SSS/DDUV/LPAD - Liver and pancreas differentiation, UCL - SSS/IREC/GAEN - Pôle d'Hépato-gastro-entérologie, UCL - (SLuc) Service de gastro-entérologie, Basic (bio-) Medical Sciences, Translational Liver Cell Biology, and Liver Cell Biology
- Subjects
0301 basic medicine ,Genetically modified mouse ,Opn ,Mutant ,knockout ,Biology ,Ck19 ,03 medical and health sciences ,0302 clinical medicine ,mouse liver ,lineage tracing ,medicine ,Recombinase ,Ectopic recombination ,cholangiocytes ,lcsh:QH301-705.5 ,bile duct cells ,Liver injury ,Bile duct ,Liver cell ,Cre ,General Medicine ,medicine.disease ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,lcsh:Biology (General) ,030211 gastroenterology & hepatology ,Tamoxifen ,medicine.drug - Abstract
Inducible cyclization recombinase (Cre) transgenic mouse strains are powerful tools for cell lineage tracing and tissue-specific knockout experiments. However, low efficiency or leaky expression can be important pitfalls. Here, we compared the efficiency and specificity of two commonly used cholangiocyte-specific Cre drivers, the Opn-iCreERT2 and Ck19-CreERT drivers, using a tdTomato reporter strain. We found that Opn-iCreERT2 triggered recombination of the tdTomato reporter in 99.9% of all cholangiocytes while Ck19-CreERT only had 32% recombination efficiency after tamoxifen injection. In the absence of tamoxifen, recombination was also induced in 2% of cholangiocytes for the Opn-iCreERT2 driver and in 13% for the Ck19-CreERT driver. For both drivers, Cre recombination was highly specific for cholangiocytes since recombination was rare in other liver cell types. Toxic liver injury ectopically activated Opn-iCreERT2 but not Ck19-CreERT expression in hepatocytes. However, ectopic recombination in hepatocytes could be avoided by applying a three-day long wash-out period between tamoxifen treatment and toxin injection. Therefore, the Opn-iCreERT2 driver is best suited for the generation of mutant bile ducts, while the Ck19-CreERT driver has near absolute specificity for bile duct cells and is therefore favorable for lineage tracing experiments.
- Published
- 2019
7. YAP and TAZ Heterogeneity in Primary Liver Cancer: An Analysis of Its Prognostic and Diagnostic Role
- Author
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Diethard Monbaliu, Chris Verslype, Guy Rens, Ruchan Karaman, Van, Haele, M, Olivier Govaere, Georg Halder, J Snoeck, Iván M. Moya, Baki Topal, Tania Roskams, and Frederik Nevens
- Subjects
0301 basic medicine ,Male ,TAZ ,Carcinogenesis ,Cell ,Kaplan-Meier Estimate ,medicine.disease_cause ,Hepatic progenitor cells ,lcsh:Chemistry ,Cholangiocarcinoma ,0302 clinical medicine ,Cytosol ,Expression pattern ,Keratin ,Medicine ,HCC ,lcsh:QH301-705.5 ,Spectroscopy ,chemistry.chemical_classification ,cHCC-CCA ,Liver Neoplasms ,Intracellular Signaling Peptides and Proteins ,General Medicine ,Middle Aged ,Prognosis ,Computer Science Applications ,Keratin 19 ,Gene Expression Regulation, Neoplastic ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Hepatocellular carcinoma ,Female ,YAP ,Primary liver cancer ,Signal Transduction ,Carcinoma, Hepatocellular ,Catalysis ,Article ,Inorganic Chemistry ,03 medical and health sciences ,Genetic Heterogeneity ,Carcinoma ,Biomarkers, Tumor ,Humans ,Physical and Theoretical Chemistry ,CCA ,Molecular Biology ,Adaptor Proteins, Signal Transducing ,Aged ,Proportional Hazards Models ,Retrospective Studies ,Cell Nucleus ,Keratin-19 ,Hippo signaling pathway ,business.industry ,Organic Chemistry ,YAP-Signaling Proteins ,medicine.disease ,Phosphoproteins ,digestive system diseases ,030104 developmental biology ,Ki-67 Antigen ,chemistry ,lcsh:Biology (General) ,lcsh:QD1-999 ,Bile Duct Neoplasms ,Transcriptional Coactivator with PDZ-Binding Motif Proteins ,Cancer research ,Trans-Activators ,business ,Transcription Factors - Abstract
Primary liver cancer comprises a diverse group of liver tumors. The heterogeneity of these tumors is seen as one of the obstacles to finding an effective therapy. The Hippo pathway, with its downstream transcriptional co-activator Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ), has a decisive role in the carcinogenesis of primary liver cancer. Therefore, we examined the expression pattern of YAP and TAZ in 141 patients with hepatocellular carcinoma keratin 19 positive (HCC K19+), hepatocellular carcinoma keratin 19 negative (HCC K19&minus, ), combined hepatocellular&ndash, cholangiocarcinoma carcinoma (cHCC-CCA), or cholangiocarcinoma (CCA). All cHCC-CCA and CCA patients showed high expression levels for YAP and TAZ, while only some patients of the HCC group were positive. Notably, we found that a histoscore of both markers is useful in the challenging diagnosis of cHCC-CCA. In addition, positivity for YAP and TAZ was observed in the hepatocellular and cholangiocellular components of cHCC-CCA, which suggests a single cell origin in cHCC-CCA. Within the K19&minus, HCC group, our results demonstrate that the expression of YAP is a statistically significant predictor of poor prognosis when observed in the cytoplasm. Nuclear expression of TAZ is an even more specific and independent predictor of poor disease-free survival and overall survival of K19&minus, HCC patients. Our results thus identify different levels of YAP/TAZ expression in various liver cancers that can be used for diagnostics.
- Published
- 2019
8. Functionality based method for simultaneous isolation of rodent hepatic sinusoidal cells
- Author
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Inge Mannaerts, Cristina Ionica Øie, L.A. van Grunsven, L. Stradiot, Iván M. Moya, Stefaan Verhulst, T. Roosens, Georg Halder, Liver Cell Biology, Faculty of Medicine and Pharmacy, Basic (bio-) Medical Sciences, and Translational Liver Cell Biology
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,medicine.drug_class ,Liver cytology ,IgG ,FACS ,KUPFFER CELLS ,Biophysics ,Bioengineering ,Cell Separation ,Biology ,Chronic liver disease ,Statistics, Nonparametric ,Rats, Sprague-Dawley ,Biomaterials ,03 medical and health sciences ,Mice ,0302 clinical medicine ,In vivo ,medicine ,Animals ,Liver sinusoidal endothelial cells ,Retinoid ,Analysis of Variance ,Mice, Inbred BALB C ,Liver cell ,Cell sorting ,medicine.disease ,Flow Cytometry ,Immunohistochemistry ,In vitro ,Coculture Techniques ,Cell biology ,Rats ,Mice, Inbred C57BL ,030104 developmental biology ,Liver ,Mechanics of Materials ,Hepatic stellate cell ,Hepatocytes ,Ceramics and Composites ,030211 gastroenterology & hepatology ,Scavenging ,hepatic stellate cells - Abstract
Chronic liver disease is the result of long term exposure to viruses or toxins such as alcohol, fat and drugs, and forms the basis for the development of liver fibrosis and primary liver cancer. In vitro and in vivo models are key to study the pathways involved in chronic liver disease and for the development of therapeutics. 3D co-culture systems are becoming the in vitro standard, which requires freshly isolated primary hepatic cells. We developed a novel isolation method to simultaneously isolate liver sinusoidal endothelial cells (LSECs), Kupffer cells (KCs) and hepatic stellate cells (HSCs). The method exploits the scavenging activity of LSECs, the phagocytic capacity of KCs and the retinoid content of HSCs in vivo to enable direct processing by fluorescence-activated cell sorting without additional antibody binding and washing steps. UFACS3, for UV-FACS-based isolation of 3 non-parenchymal liver cell types, yields functional and pure LSECs (98 ± 1%), KCs (98 ± 1%) and HSCs (97 ± 3%), with less hands-on time from healthy and diseased rodent livers. This novel approach allows a fast and effective combined isolation of sinusoidal cells for further analysis.
- Published
- 2017
9. Antagonism of Nodal signaling by BMP/Smad5 prevents ectopic primitive streak formation in the mouse amnion
- Author
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Mariya P. Dobreva, F.M. Cornelis, An Zwijsen, Lieve Umans, Danny Huylebroeck, Paulo N. G. Pereira, Catherine M. Verfaillie, Susana M. Chuva de Sousa Lopes, Anne Camus, Iván M. Moya, Elke Maas, Department of Molecular and Developmental Genetics (VIB11), Flanders institute of biotechnology, Laboratory of Molecular Biology (CELGEN), Catholic University of Leuven - Katholieke Universiteit Leuven (KU Leuven), Interdepartmental Stem Cell Institute Leuven, Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Department of Anatomy and Embryology, Leiden University Medical Center, Leiden University Medical Center (LUMC), Ciência e Tecnologia (FCT) [SFRH/BD/15901/2005] (GABBA program class of 2005), FCT, and Interuniversity Attraction Poles Program IUAP-6, FWO-V (G.0382.07), OT-09/053 and GOA-11/012, he KU Leuven Research Council
- Subjects
Mouse ,MESH: Bone Morphogenetic Proteins ,Nodal signaling ,Nodal ,Mice ,0302 clinical medicine ,MESH: Pregnancy ,Pregnancy ,MESH: Reverse Transcriptase Polymerase Chain Reaction ,MESH: Animals ,[SDV.BDD]Life Sciences [q-bio]/Development Biology ,In Situ Hybridization ,Smad ,0303 health sciences ,Amnion ,Primitive streak formation ,Primitive streak ,Reverse Transcriptase Polymerase Chain Reaction ,Immunohistochemistry ,Cell biology ,medicine.anatomical_structure ,Bone Morphogenetic Proteins ,embryonic structures ,Female ,Smad5 Protein ,medicine.medical_specialty ,animal structures ,Nodal Protein ,Blotting, Western ,Biology ,Bone morphogenetic protein ,Cell Line ,03 medical and health sciences ,MESH: Smad5 Protein ,MESH: In Situ Hybridization ,Internal medicine ,medicine ,Animals ,Humans ,Immunoprecipitation ,MESH: Blotting, Western ,MESH: Amnion ,Molecular Biology ,MESH: Mice ,MESH: Nodal Protein ,030304 developmental biology ,MESH: Humans ,MESH: Immunoprecipitation ,Bone Morphogenetic Protein ,MESH: Immunohistochemistry ,Embryonic stem cell ,MESH: Cell Line ,MESH: Primitive Streak ,Endocrinology ,Epiblast ,NODAL ,MESH: Female ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
The strength and spatiotemporal activity of Nodal signaling is tightly controlled in early implantation mouse embryos, including by autoregulation and feedback loops, and involves secreted and intracellular antagonists. These control mechanisms, which are established at the extra-embryonic/embryonic interfaces, are essential for anterior-posterior patterning of the epiblast and correct positioning of the primitive streak. Formation of an ectopic primitive streak, or streak expansion, has previously been reported in mutants lacking antagonists that target Nodal signaling. Here, we demonstrate that loss-of-function of a major bone morphogenetic protein (BMP) effector, Smad5, results in formation of an ectopic primitive streak-like structure in mutant amnion accompanied by ectopic Nodal expression. This suggests that BMP/Smad5 signaling contributes to negative regulation of Nodal. In cultured cells, we find that BMP-activated Smad5 antagonizes Nodal signaling by interfering with the Nodal-Smad2/4-Foxh1 autoregulatory pathway through the formation of an unusual BMP4-induced Smad complex containing Smad2 and Smad5. Quantitative expression analysis supports that ectopic Nodal expression in the Smad5 mutant amnion is induced by the Nodal autoregulatory loop and a slow positive-feedback loop. The latter involves BMP4 signaling and also induction of ectopic Wnt3. Ectopic activation of these Nodal feedback loops in the Smad5 mutant amnion results in the eventual formation of an ectopic primitive streak-like structure. We conclude that antagonism of Nodal signaling by BMP/Smad5 signaling prevents primitive streak formation in the amnion of normal mouse embryos. ispartof: Development vol:139 issue:18 pages:3343-3354 ispartof: location:England status: published
- Published
- 2012
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