Publicaciones por año
Publicaciones del Departamento de Histología y Embriología
CD300f immunoreceptor is associated with major depressive disorder and decreased microglial metabolic fitness
Proc Natl Acad Sci U S A 2020 117(12):6651-6662
Natalia Lago 1 , Fernanda N Kaufmann 2 , María Luciana Negro-Demontel 1 3 , Daniela Alí-Ruiz 1 , Gabriele Ghisleni 4 , Natalia Rego 5 , Andrea Arcas-García 6 , Nathalia Vitureira 1 7 , Karen Jansen 4 , Luciano M Souza 4 , Ricardo A Silva 4 , Diogo R Lara 8 , Bruno Pannunzio 1 3 , Juan Andrés Abin-Carriquiry 9 , Jesús Amo-Aparicio 10 , Celia Martin-Otal 6 , Hugo Naya 6 , Dorian B McGavern 11 , Joan Sayós 6 , Rubèn López-Vales 10 , Manuella P Kaster 2 , Hugo Peluffo 12 3
1 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, 11400 Montevideo, Uruguay. 2 Department of Biochemistry, Federal University of Santa Catarina, Florianópolis, 88040-900 Santa Catarina, Brazil. 3 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, 11200 Montevideo, Uruguay. 4 Department of Life and Health Sciences, Catholic University of Pelotas, 96015-560 Rio Grande do Sul, Brazil. 5 Bioinformatics Unit, Institut Pasteur de Montevideo, 11400 Montevideo, Uruguay. 6 Immune Regulation and Immunotherapy Group, CIBBIM-Nanomedicine, Vall d'Hebrón Institut de Recerca, Universitat Autònoma de Barcelona, 08193 Barcelona, Spain. 7 Department of Physiology, Facultad de Medicina, Universidad de la República, 11200 Montevideo, Uruguay. 8 Department of Cellular and Molecular Biology, Pontifical Catholic University of Rio Grande do Sul, 90619-900 Porto Alegre, Brazil. 9 Instituto de Investigaciones Biológicas Clemente Estable, 11600 Montevideo, Uruguay. 10 Departament de Biologia Cel·lular, Fisiologia i Immunologia, Institut de Neurociències, Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain. 11 Viral Immunology and Intravital Imaging Section, National Institute for Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892. 12 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, 11400 Montevideo, Uruguay; hugo.peluffo@pasteur.edu.uy.
DOI: 10.1073/pnas.1911816117
PMID: 32152116
Pubmed: https://pubmed.ncbi.nlm.nih.gov/32152116
Texto completo: https://www.pnas.org/doi/full/10.1073/pnas.1911816117
Abstract:
A role for microglia in neuropsychiatric diseases, including major depressive disorder (MDD), has been postulated. Regulation of microglial phenotype by immune receptors has become a central topic in many neurological conditions. We explored preclinical and clinical evidence for the role of the CD300f immune receptor in the fine regulation of microglial phenotype and its contribution to MDD. We found that a prevalent nonsynonymous single-nucleotide polymorphism (C/T, rs2034310) of the human CD300f receptor cytoplasmic tail inhibits the protein kinase C phosphorylation of a threonine and is associated with protection against MDD, mainly in women. Interestingly, CD300f-/- mice displayed several characteristic MDD traits such as augmented microglial numbers, increased interleukin 6 and interleukin 1 receptor antagonist messenger RNA, alterations in synaptic strength, and noradrenaline-dependent and persistent depressive-like and anhedonic behaviors in females. This behavioral phenotype could be potentiated inducing the lipopolysaccharide depression model. RNA sequencing and biochemical studies revealed an association with impaired microglial metabolic fitness. In conclusion, we report a clear association that links the function of the CD300f immune receptor with MDD in humans, depressive-like and anhedonic behaviors in female mice, and altered microglial metabolic reprogramming.
Signaling pathways in cytoskeletal responses to plasma membrane depolarization in corneal endothelial cells
J Cell Physiol 2020 235(3):2947-2962
Frances Evans 1 , Julio A Hernández 2 , Silvia Chifflet 3
1 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 2 Sección Biofísica, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay. 3 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
DOI: 10.1002/jcp.29200
PMID: 31535377
Pubmed: https://pubmed.ncbi.nlm.nih.gov/31535377
Texto completo: https://doi.org/10.1002/jcp.29200
Abstract:
In previous work, we reported that plasma membrane potential depolarization (PMPD) provokes cortical F-actin remodeling in bovine corneal endothelial (BCE) cells in culture, which eventually leads to the appearance of intercellular gaps. In kidney epithelial cells it has been shown that PMPD determines an extracellular-signal-regulated kinase (ERK)/Rho-dependent increase in diphosphorylated myosin light chain (ppMLC). The present study investigated the signaling pathways involved in the response of BCE cells to PMPD. Differently to renal epithelial cells, we observed that PMPD leads to a decrease in monophosphorylated MLC (pMLC) without affecting diphosphorylated MLC. Also, that the pMLC reduction is a consequence of cyclic adenosine 3',5'-monophosphate (cAMP)/protein kinase A (PKA) activation. In addition, we found evidence that the cAMP increase mostly depends on soluble adenylyl cyclase activity. Inhibition of this enzyme reduces the effect of PMPD on the cAMP rise, F-actin remodeling, and pMLC decrease. No changes in phosho-ERK were observed, although we could determine that RhoA undergoes activation. Our results suggested that active RhoA is not involved in the intercellular gap formation. Overall, the findings of this study support the view that, differently to renal epithelial cells, in BCE cells PMPD determines cytoskeletal reorganization via activation of the cAMP/PKA pathway.
Signaling pathways in cytoskeletal responses to plasma membrane depolarization in corneal endothelial cells
J Cell Physiol 2020 235(3):2947-2962
Frances Evans 1 , Julio A Hernández 2 , Silvia Chifflet 3
1 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 2 Sección Biofísica, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay. 3 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
DOI: 10.1002/jcp.29200
PMID: 31535377
Pubmed: https://pubmed.ncbi.nlm.nih.gov/31535377
Texto completo: https://doi.org/10.1002/jcp.29200
Abstract:
In previous work, we reported that plasma membrane potential depolarization (PMPD) provokes cortical F-actin remodeling in bovine corneal endothelial (BCE) cells in culture, which eventually leads to the appearance of intercellular gaps. In kidney epithelial cells it has been shown that PMPD determines an extracellular-signal-regulated kinase (ERK)/Rho-dependent increase in diphosphorylated myosin light chain (ppMLC). The present study investigated the signaling pathways involved in the response of BCE cells to PMPD. Differently to renal epithelial cells, we observed that PMPD leads to a decrease in monophosphorylated MLC (pMLC) without affecting diphosphorylated MLC. Also, that the pMLC reduction is a consequence of cyclic adenosine 3',5'-monophosphate (cAMP)/protein kinase A (PKA) activation. In addition, we found evidence that the cAMP increase mostly depends on soluble adenylyl cyclase activity. Inhibition of this enzyme reduces the effect of PMPD on the cAMP rise, F-actin remodeling, and pMLC decrease. No changes in phosho-ERK were observed, although we could determine that RhoA undergoes activation. Our results suggested that active RhoA is not involved in the intercellular gap formation. Overall, the findings of this study support the view that, differently to renal epithelial cells, in BCE cells PMPD determines cytoskeletal reorganization via activation of the cAMP/PKA pathway.
Spotlight edition on South America
Gene Ther 2020 27(1-2):1
Ursula Matte 1 , Hugo Peluffo 2
1 Post-Graduation Program on Genetics and Molecular Biology, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2350, Porto Alegre, 90035-903, Brazil. umatte@hcpa.edu.br. 2 Department of Histology and Embryology, Facultad de Medicina, Universidad de la República, Mataojo 2020, Montevideo, 11400, Uruguay.
DOI: 10.1038/s41434-020-0129-9
PMID: 32099107
Pubmed: https://pubmed.ncbi.nlm.nih.gov/32099107
Texto completo: https://doi.org/10.1038/s41434-020-0129-9
Abstract:
Safe and neuroprotective vectors for long-term traumatic brain injury gene therapy
Gene Ther 2020 27(1-2):96-103
Daniela Blanco-Ocampo 1 2 , Fabio Andrés Cawen 3 , Luis Angel Álamo-Pindado 1 3 , María Luciana Negro-Demontel 1 3 , Hugo Peluffo 4 5
1 Department of Histology and Embryology, Faculty of Medicine, Universidad de la República, Montevideo, Uruguay. 2 Department of Physiopathology, Faculty of Medicine, Universidad de la República, Montevideo, Uruguay. 3 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. 4 Department of Histology and Embryology, Faculty of Medicine, Universidad de la República, Montevideo, Uruguay. hugo.peluffo@pasteur.edu.uy. 5 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. hugo.peluffo@pasteur.edu.uy.
DOI: 10.1038/s41434-019-0073-8
PMID: 30926962
Pubmed: https://pubmed.ncbi.nlm.nih.gov/30926962
Texto completo: https://doi.org/10.1038/s41434-019-0073-8
Abstract:
Traumatic brain injury (TBI) is a complex and progressive brain injury with no approved treatments that needs both short- and long-term therapeutic strategies to cope with the variety of physiopathological mechanisms involved. In particular, neuroinflammation is a key process modulating TBI outcome, and the potentiation of these mechanisms by pro-inflammatory gene therapy vectors could contribute to the injury progression. Here, we evaluate in the controlled cortical impact model of TBI, the safety of integrative-deficient lentiviral vectors (IDLVs) or the non-viral HNRK recombinant modular protein/DNA nanovector. These two promising vectors display different tropisms, transduction efficiencies, short- or long-term transduction or inflammatory activation profile. We show that the brain intraparenchymal injection of these vectors overexpressing green fluorescent protein after a CCI is not neurotoxic, and interestingly, can decrease the short-term sensory neurological deficits, and diminish the brain tissue loss at 90 days post lesion (dpl). Moreover, only IDLVs were able to mitigate the memory deficits elicited by a CCI. These vectors did not alter the microglial or astroglial reactivity at 90 dpl, suggesting that they do not potentiate the on-going neuroinflammation. Taken together, these data suggest that both types of vectors could be interesting tools for the design of gene therapy strategies targeting immediate or long-term neuropathological mechanisms of TBI.
Leukocyte Profiles Reflect Geographic Range Limits in a Widespread Neotropical Bat
Integr Comp Biol 2019 59(5):1176-1189
Daniel J Becker 1 2 3 , Cecilia Nachtmann 1 , Hernan D Argibay 4 , Germán Botto 5 6 , Marina Escalera-Zamudio 7 8 , Jorge E Carrera 9 10 , Carlos Tello 11 12 , Erik Winiarski 13 , Alex D Greenwood 7 14 , Maria L Méndez-Ojeda 15 , Elizabeth Loza-Rubio 16 , Anne Lavergne 17 , Benoit de Thoisy 17 , Gábor Á Czirják 7 , Raina K Plowright 5 , Sonia Altizer 1 2 , Daniel G Streicker 1 18 19
1 Odum School of Ecology, University of Georgia, Athens, GA 30602, USA. 2 Center for the Ecology of Infectious Disease, University of Georgia, Athens, GA 30602, USA. 3 Department of Biology, Indiana University, Bloomington, IN 47405, USA. 4 Departamento de Ecología, Genética y Evolución, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires C1428EGA, Argentina. 5 Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59715, USA. 6 Departamento de Metodos Cuantitativos, Facultad de Medicina, Universidad de la República, Montevideo 11800, Uruguay. 7 Department of Wildlife Diseases, Leibniz Institute for Zoo and Wildlife Research, Berlin 10315, Germany. 8 Department of Zoology, University of Oxford, Oxford OX1 3SY, UK. 9 Facultad de Ciencias, Universidad Nacional de Piura, Piura 20009, Peru. 10 Programa de Conservación de Murciélagos de Perú, Piura Lima-1, Peru. 11 Association for the Conservation and Development of Natural Resources, Lima 15037, Peru. 12 Yunkawasi, Lima 15049, Peru. 13 Departamento de Histología, Facultad de Medicina, Universidad de la República, Montevideo 11800, Uruguay. 14 Department of Veterinary Medicine, Freie Universität Berlin, Berlin 14163, Germany. 15 Facultad de Medicina Veterinaria y Zootecnia, Universidad Veracruzana, Veracruz 91710, Mexico. 16 Centro Nacional de Investigación Disciplinaria en Microbiología Animal, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Mexico City 05110, Mexico. 17 Laboratoire des Interactions Virus-Hôtes, Institut Pasteur de la Guyane, Cayenne, French Guiana F-97300, France. 18 Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK. 19 MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, UK.
DOI: 10.1093/icb/icz007
PMID: 30873523
Pubmed: https://pubmed.ncbi.nlm.nih.gov/30873523
Texto completo: https://academic.oup.com/icb/article-lookup/doi/10.1093/icb/icz007
Abstract:
Quantifying how the environment shapes host immune defense is important for understanding which wild populations may be more susceptible or resistant to pathogens. Spatial variation in parasite risk, food and predator abundance, and abiotic conditions can each affect immunity, and these factors can also manifest at both local and biogeographic scales. Yet identifying predictors and the spatial scale of their effects is limited by the rarity of studies that measure immunity across many populations of broadly distributed species. We analyzed leukocyte profiles from 39 wild populations of the common vampire bat (Desmodus rotundus) across its wide geographic range throughout the Neotropics. White blood cell differentials varied spatially, with proportions of neutrophils and lymphocytes varying up to six-fold across sites. Leukocyte profiles were spatially autocorrelated at small and very large distances, suggesting that local environment and large-scale biogeographic factors influence cellular immunity. Generalized additive models showed that bat populations closer to the northern and southern limits of the species range had more neutrophils, monocytes, and basophils, but fewer lymphocytes and eosinophils, than bats sampled at the core of their distribution. Habitats with access to more livestock also showed similar patterns in leukocyte profiles, but large-scale patterns were partly confounded by time between capture and sampling across sites. Our findings suggest that populations at the edge of their range experience physiologically limiting conditions that predict higher chronic stress and greater investment in cellular innate immunity. High food abundance in livestock-dense habitats may exacerbate such conditions by increasing bat density or diet homogenization, although future spatially and temporally coordinated field studies with common protocols are needed to limit sampling artifacts. Systematically assessing immune function and response over space will elucidate how environmental conditions influence traits relevant to epidemiology and help predict disease risks with anthropogenic disturbance, land conversion, and climate change.
Glutaric Acid Affects Pericyte Contractility and Migration: Possible Implications for GA-I Pathogenesis
Mol Neurobiol 2019 56(11):7694-7707
Eugenia Isasi 1 2 , Nils Korte 3 , Verónica Abudara 4 , David Attwell 3 , Silvia Olivera-Bravo 5
1 Neurobiología Celular y Molecular, Instituto Clemente Estable (IIBCE), 3318, Italia Av, 11600, Montevideo, Uruguay. 2 Departmento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 3 Department of Neuroscience, Physiology and Pharmacology, University College London (UCL), London, UK. 4 Departmento de Fisiología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 5 Neurobiología Celular y Molecular, Instituto Clemente Estable (IIBCE), 3318, Italia Av, 11600, Montevideo, Uruguay. solivera2011@gmail.com.
DOI: 10.1007/s12035-019-1620-4
PMID: 31104295
Pubmed: https://pubmed.ncbi.nlm.nih.gov/31104295
Texto completo: https://dx.doi.org/10.1007/s12035-019-1620-4
Abstract:
Glutaric acidemia I (GA-I) is an inherited neurometabolic childhood disease characterized by bilateral striatal neurodegeneration upon brain accumulation of millimolar concentrations of glutaric acid (GA) and related metabolites. Vascular dysfunction, including abnormal cerebral blood flow and blood-brain barrier damage, is an early pathological feature in GA-I, although the affected cellular targets and underlying mechanisms remain unknown. In the present study, we have assessed the effects of GA on capillary pericyte contractility in cerebral cortical slices and pericyte cultures, as well as on the survival, proliferation, and migration of cultured pericytes. GA induced a significant reduction in capillary diameter at distances up to ~ 10 μm from the center of pericyte somata. However, GA did not affect the contractility of cultured pericytes, suggesting that the response elicited in slices may involve GA evoking pericyte contraction by acting on other cellular components of the neurovascular unit. Moreover, GA indirectly inhibited migration of cultured pericytes, an effect that was dependent on soluble glial factors since it was observed upon application of conditioned media from GA-treated astrocytes (CM-GA), but not upon direct GA addition to the medium. Remarkably, CM-GA showed increased expression of cytokines and growth factors that might mediate the effects of increased GA levels not only on pericyte migration but also on vascular permeability and angiogenesis. These data suggest that some effects elicited by GA might be produced by altering astrocyte-pericyte communication, rather than directly acting on pericytes. Importantly, GA-evoked alteration of capillary pericyte contractility may account for the reduced cerebral blood flow observed in GA-I patients.
A novel form of Deleted in breast cancer 1 (DBC1) lacking the N-terminal domain does not bind SIRT1 and is dynamically regulated in vivo
Sci Rep 2019 9(1):14381
Leonardo Santos 1 , Laura Colman 1 , Paola Contreras 1 2 , Claudia C Chini 3 , Adriana Carlomagno 1 , Alejandro Leyva 4 , Mariana Bresque 1 , Inés Marmisolle 5 , Celia Quijano 5 , Rosario Durán 4 , Florencia Irigoín 6 7 , Victoria Prieto-Echagüe 6 , Mikkel H Vendelbo 8 9 , José R Sotelo-Silveira 10 , Eduardo N Chini 3 , Jose L Badano 6 , Aldo J Calliari 1 11 , Carlos Escande 12
1 Laboratory of Metabolic Diseases and Aging, INDICyO Program, Institut Pasteur de Montevideo, Montevideo, Uruguay. 2 Departamento de Fisiología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 3 Signal Transduction and Molecular Nutrition Laboratory, Kogod Aging Center, Department of Anesthesiology and Perioperative Medicine, Mayo Clinic College of Medicine, Rochester, USA. 4 Analytical Biochemistry and Proteomics Unit, Institut Pasteur de Montevideo and Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay. 5 Departamento de Bioquímica and Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 6 Human Molecular Genetics, INDICyO Program, Institut Pasteur de Montevideo, Montevideo, Uruguay. 7 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 8 Department of Nuclear Medicine and PET Centre, Aarhus University Hospital, Aarhus, Denmark. 9 Department of Biomedicine, Aarhus University, Aarhus, Denmark. 10 Department of Genomics, Instituto de Investigaciones Biológicas Clemente Estable, and Laboratory of Molecular Interactions, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay. 11 Area Biofísica, Departamento de Biología Celular y Molecular, Facultad de Veterinaria, Universidad de la República, Montevideo, Uruguay. 12 Laboratory of Metabolic Diseases and Aging, INDICyO Program, Institut Pasteur de Montevideo, Montevideo, Uruguay. escande@pasteur.edu.uy.
DOI: 10.1038/s41598-019-50789-7
PMID: 31591441
Pubmed: https://pubmed.ncbi.nlm.nih.gov/31591441
Texto completo: https://doi.org/10.1038/s41598-019-50789-7
Abstract:
The protein Deleted in Breast Cancer-1 is a regulator of several transcription factors and epigenetic regulators, including HDAC3, Rev-erb-alpha, PARP1 and SIRT1. It is well known that DBC1 regulates its targets, including SIRT1, by protein-protein interaction. However, little is known about how DBC1 biological activity is regulated. In this work, we show that in quiescent cells DBC1 is proteolytically cleaved, producing a protein (DN-DBC1) that misses the S1-like domain and no longer binds to SIRT1. DN-DBC1 is also found in vivo in mouse and human tissues. Interestingly, DN-DBC1 is cleared once quiescent cells re-enter to the cell cycle. Using a model of liver regeneration after partial hepatectomy, we found that DN-DBC1 is down-regulated in vivo during regeneration. In fact, WT mice show a decrease in SIRT1 activity during liver regeneration, coincidentally with DN-DBC1 downregulation and the appearance of full length DBC1. This effect on SIRT1 activity was not observed in DBC1 KO mice. Finally, we found that DBC1 KO mice have altered cell cycle progression and liver regeneration after partial hepatectomy, suggesting that DBC1/DN-DBC1 transitions play a role in normal cell cycle progression in vivo after cells leave quiescence. We propose that quiescent cells express DN-DBC1, which either replaces or coexist with the full-length protein, and that restoring of DBC1 is required for normal cell cycle progression in vitro and in vivo. Our results describe for the first time in vivo a naturally occurring form of DBC1, which does not bind SIRT1 and is dynamically regulated, thus contributing to redefine the knowledge about its function.
A novel form of Deleted in breast cancer 1 (DBC1) lacking the N-terminal domain does not bind SIRT1 and is dynamically regulated in vivo
Sci Rep 2019 9(1):14381
Leonardo Santos 1 , Laura Colman 1 , Paola Contreras 1 2 , Claudia C Chini 3 , Adriana Carlomagno 1 , Alejandro Leyva 4 , Mariana Bresque 1 , Inés Marmisolle 5 , Celia Quijano 5 , Rosario Durán 4 , Florencia Irigoín 6 7 , Victoria Prieto-Echagüe 6 , Mikkel H Vendelbo 8 9 , José R Sotelo-Silveira 10 , Eduardo N Chini 3 , Jose L Badano 6 , Aldo J Calliari 1 11 , Carlos Escande 12
1 Laboratory of Metabolic Diseases and Aging, INDICyO Program, Institut Pasteur de Montevideo, Montevideo, Uruguay. 2 Departamento de Fisiología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 3 Signal Transduction and Molecular Nutrition Laboratory, Kogod Aging Center, Department of Anesthesiology and Perioperative Medicine, Mayo Clinic College of Medicine, Rochester, USA. 4 Analytical Biochemistry and Proteomics Unit, Institut Pasteur de Montevideo and Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay. 5 Departamento de Bioquímica and Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 6 Human Molecular Genetics, INDICyO Program, Institut Pasteur de Montevideo, Montevideo, Uruguay. 7 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 8 Department of Nuclear Medicine and PET Centre, Aarhus University Hospital, Aarhus, Denmark. 9 Department of Biomedicine, Aarhus University, Aarhus, Denmark. 10 Department of Genomics, Instituto de Investigaciones Biológicas Clemente Estable, and Laboratory of Molecular Interactions, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay. 11 Area Biofísica, Departamento de Biología Celular y Molecular, Facultad de Veterinaria, Universidad de la República, Montevideo, Uruguay. 12 Laboratory of Metabolic Diseases and Aging, INDICyO Program, Institut Pasteur de Montevideo, Montevideo, Uruguay. escande@pasteur.edu.uy.
DOI: 10.1038/s41598-019-50789-7
PMID: 31591441
Pubmed: https://pubmed.ncbi.nlm.nih.gov/31591441
Texto completo: https://doi.org/10.1038/s41598-019-50789-7
Abstract:
The protein Deleted in Breast Cancer-1 is a regulator of several transcription factors and epigenetic regulators, including HDAC3, Rev-erb-alpha, PARP1 and SIRT1. It is well known that DBC1 regulates its targets, including SIRT1, by protein-protein interaction. However, little is known about how DBC1 biological activity is regulated. In this work, we show that in quiescent cells DBC1 is proteolytically cleaved, producing a protein (DN-DBC1) that misses the S1-like domain and no longer binds to SIRT1. DN-DBC1 is also found in vivo in mouse and human tissues. Interestingly, DN-DBC1 is cleared once quiescent cells re-enter to the cell cycle. Using a model of liver regeneration after partial hepatectomy, we found that DN-DBC1 is down-regulated in vivo during regeneration. In fact, WT mice show a decrease in SIRT1 activity during liver regeneration, coincidentally with DN-DBC1 downregulation and the appearance of full length DBC1. This effect on SIRT1 activity was not observed in DBC1 KO mice. Finally, we found that DBC1 KO mice have altered cell cycle progression and liver regeneration after partial hepatectomy, suggesting that DBC1/DN-DBC1 transitions play a role in normal cell cycle progression in vivo after cells leave quiescence. We propose that quiescent cells express DN-DBC1, which either replaces or coexist with the full-length protein, and that restoring of DBC1 is required for normal cell cycle progression in vitro and in vivo. Our results describe for the first time in vivo a naturally occurring form of DBC1, which does not bind SIRT1 and is dynamically regulated, thus contributing to redefine the knowledge about its function.
Mitofusins modulate the increase in mitochondrial length, bioenergetics and secretory phenotype in therapy-induced senescent melanoma cells
Biochem J 2019 476(17):2463-2486
Jennyfer Martínez 1 , Doménica Tarallo 1 , Laura Martínez-Palma 2 , Sabina Victoria 3 , Mariana Bresque 4 , Sebastián Rodríguez-Bottero 2 , Inés Marmisolle 1 , Carlos Escande 4 , Patricia Cassina 2 , Gabriela Casanova 5 , Mariela Bollati-Fogolín 3 , Caroline Agorio 6 , María Moreno 7 , Celia Quijano 8
1 Centro de Investigaciones Biomédicas (CEINBIO) and Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 2 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 3 Cell Biology Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay. 4 Metabolic Diseases and Aging Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. 5 Unidad de Microscopía Electrónica de Transmisión, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay. 6 Cátedra de Dermatología, Hospital de Clínicas Manuel Quintela, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 7 Laboratory for Vaccine Research, Departamento de Desarrollo Biotecnológico, Instituto de Higiene, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 8 Centro de Investigaciones Biomédicas (CEINBIO) and Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay celiq@fmed.edu.uy celia.quijano@gmail.com.
DOI: 10.1042/BCJ20190405
PMID: 31431479
Pubmed: https://pubmed.ncbi.nlm.nih.gov/31431479
Texto completo: https://portlandpress.com/biochemj/article-lookup/doi/10.1042/BCJ20190405
Abstract:
Cellular senescence is an endpoint of chemotherapy, and targeted therapies in melanoma and the senescence-associated secretory phenotype (SASP) can affect tumor growth and microenvironment, influencing treatment outcomes. Metabolic interventions can modulate the SASP, and an enhanced mitochondrial energy metabolism supports resistance to therapy in melanoma cells. Herein, we assessed the mitochondrial function of therapy-induced senescent melanoma cells obtained after exposing the cells to temozolomide (TMZ), a methylating chemotherapeutic agent. Senescence induction in melanoma was accompanied by a substantial increase in mitochondrial basal, ATP-linked, and maximum respiration rates and in coupling efficiency, spare respiratory capacity, and respiratory control ratio. Further examinations revealed an increase in mitochondrial mass and length. Alterations in mitochondrial function and morphology were confirmed in isolated senescent cells, obtained by cell-size sorting. An increase in mitofusin 1 and 2 (MFN1 and 2) expression and levels was observed in senescent cells, pointing to alterations in mitochondrial fusion. Silencing mitofusin expression with short hairpin RNA (shRNA) prevented the increase in mitochondrial length, oxygen consumption rate and secretion of interleukin 6 (IL-6), a component of the SASP, in melanoma senescent cells. Our results represent the first in-depth study of mitochondrial function in therapy-induced senescence in melanoma. They indicate that senescence increases mitochondrial mass, length and energy metabolism; and highlight mitochondria as potential pharmacological targets to modulate senescence and the SASP.