39 results on '"Porreca, I."'
Search Results
2. Molecular targets of developmental exposure to bisphenol A in diabesity: a focus on endoderm‐derived organs
- Author
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Porreca, I., Ulloa‐Severino, L., Almeida, P., Cuomo, D., Nardone, A., Falco, G., Mallardo, M., and Ambrosino, C.
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- 2017
- Full Text
- View/download PDF
3. Localization of BDNF expression in the developing brain of zebrafish
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De Felice, E., Porreca, I., Alleva, E., De Girolamo, P., Ambrosino, C., Ciriaco, E., Germanà, A., and Sordino, P.
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- 2014
- Full Text
- View/download PDF
4. Molecular targets of developmental exposure to bisphenol A in diabesity: a focus on endoderm-derived organs
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Porreca, I, Ulloa Severino, L, Almeida, P, Cuomo, D, NARDONE, Antonio, FALCO, Geppino, MALLARDO, MASSIMO, Ambrosino, C., Porreca, I, ULLOA SEVERINO, Luisa, Almeida, P., Cuomo, D., Nardone, A., Falco, G., Mallardo, M., Ambrosino, C., Ulloa Severino, L, Almeida, P, Cuomo, D, Nardone, Antonio, Falco, Geppino, and Mallardo, Massimo
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Male ,endoderm-derived organs ,Endocrinology, Diabetes and Metabolism ,Bisphenol A ,epigenetics ,perinatal exposure ,Public Health, Environmental and Occupational Health ,Thyroid Gland ,Endocrinology ,Metabolic Diseases ,Phenols ,Animals ,Humans ,Obesity ,Benzhydryl Compounds ,Lung ,Pancreas ,Environmental and Occupational Health ,Endoderm ,Prostate ,Diabetes and Metabolism ,Gastrointestinal Tract ,Disease Models, Animal ,Diabetes Mellitus, Type 2 ,Liver ,Public Health ,endoderm-derived organ ,epigenetic - Abstract
Several studies associate foetal human exposure to bisphenol A (BPA) to metabolic/endocrine diseases, mainly diabesity. They describe the role of BPA in the disruption of pancreatic beta cell, adipocyte and hepatocyte functions. Indeed, the complexity of the diabesity phenotype is due to the involvement of different endoderm-derived organs, all targets of BPA. Here, we analyse this point delineating a picture of different mechanisms of BPA toxicity in endoderm-derived organs leading to diabesity. Moving from epidemiological data, we summarize the in vivo experimental data of the BPA effects on endoderm-derived organs (thyroid, pancreas, liver, gut, prostate and lung) after prenatal exposure. Mainly, we gather molecular data evidencing harmful effects at low-dose exposure, pointing to the risk to human health. Although the fragmentation of molecular data does not allow a clear conclusion to be drawn, the present work indicates that the developmental exposure to BPA represents a risk for endoderm-derived organs development as it deregulates the gene expression from the earliest developmental stages. A more systematic analysis of BPA impact on the transcriptomes of endoderm-derived organs is still missing. Here, we suggest in vitro toxicogenomics approaches as a tool for the identification of common mechanisms of BPA toxicity leading to the diabesity in organs having the same developmental origin.
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- 2016
5. Evaluation of low doses BPA-induced perturbation of glycemia by toxicogenomics points to a primary role of pancreatic islets and to the mechanism of toxicity
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Carchia, E, primary, Porreca, I, additional, Almeida, P J, additional, D'Angelo, F, additional, Cuomo, D, additional, Ceccarelli, M, additional, De Felice, M, additional, Mallardo, M, additional, and Ambrosino, C, additional
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- 2015
- Full Text
- View/download PDF
6. Uncovering new genes involved in thyroid gland development
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Porreca, I., Fagman, H., De Felice, E., Germana', Antonino, De Felice, M., Sordino, P., and Di Lauro, R.
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- 2009
7. Genetic background and window of exposure contribute to thyroid dysfunction promoted by low-dose exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin in mice
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Immacolata Porreca, Luca Roberto, Concetta Ambrosino, Mario De Felice, Maria Marotta, Emanuele Carchia, Nicola Antonino Russo, Massimo Mallardo, Filomena Russo, Carla Reale, Reale, C., Porreca, I., Russo, F., Marotta, M., Roberto, L., Russo, N. A., Carchia, E., Mallardo, Massimo., De Felice, M., and Ambrosino, C.
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0301 basic medicine ,Male ,Polychlorinated Dibenzodioxins ,Time Factors ,Thyroid Nuclear Factor 1 ,Thyroid Gland ,lcsh:Medicine ,Haploinsufficiency ,Mice ,NF-KappaB Inhibitor alpha ,Gene expression ,Medicine ,lcsh:Science ,Sex Characteristics ,Multidisciplinary ,Thyroid ,Phenotype ,Dose–response relationship ,medicine.anatomical_structure ,Polychlorinated Dibenzodioxin ,Female ,Thyroid function ,Signal Transduction ,medicine.medical_specialty ,endocrine system ,Time Factor ,Article ,Cell Line ,03 medical and health sciences ,PAX8 Transcription Factor ,Hypothyroidism ,In vivo ,Internal medicine ,Animals ,Dose-Response Relationship, Drug ,business.industry ,Animal ,lcsh:R ,Transcription Factor RelA ,Sex Characteristic ,8-tetrachlorodibenzo-p-dioxin ,genetic ,Pax8 ,Nkx2-1 ,hypothyroidism ,humans ,Furthermore ,030104 developmental biology ,Endocrinology ,lcsh:Q ,Tumor Suppressor Protein p53 ,business ,PAX8 - Abstract
Genetic and environmental factors contribute to thyroid diseases. Although still debated, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is thought to induce thyroid dysfunction in humans and rodents. The data here reported point out the contribution of the exposure window and genetic background in mediating the low-dose TCDD effects on thyroid. Indeed, early (from E0.5 to PND30) and low-dose (0,001 μg/kg/day) TCDD exposure reduced the circulating fT4 and altered the expression of thyroid specific transcripts. The role of genetic components was estimated monitoring the same markers in Pax8+/− and Nkx2-1+/− mice, susceptible to thyroid dysfunction, exposed to 0, 1 μg/kg/day TCDD from E15.5 to PND60. Haploinsufficiency of either Pax8 or Nkx2-1 genes exacerbated the effects of the exposure impairing the thyroid enriched mRNAs in sex dependent manner. Such effect was mediated by mechanisms involving the Nkx2-1/p53/p65/IĸBα pathway in vitro and in vivo. Foetal exposure to TCDD impaired both thyroid function and genes expression while thyroid development and differentiation did not appear significantly affected. In mouse, stronger effects were related to earlier exposure or specific genetic background such as either Pax8 or Nkx2-1 haploinsufficiency, both associated to hypothyroidism in humans. Furthermore, our data underline that long exposure time are needed to model in vitro and in vivo results.
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- 2017
8. Characterization of paralogous uncx transcription factor encoding genes in zebrafish
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Marcella Fiengo, Rita Marino, Takashi Kondo, Sylvie Maiella, Antonio Emidio Fortunato, Fernanda Langellotto, Raffaella De Paolo, Stephen W. Wilson, Giulia Fasano, Filomena Ristoratore, Lorella M.T. Canzoniero, Valeria Nittoli, Alessandra Gentile, Paolo Sordino, Ugo Coppola, Francesco Aniello, Aldo Donizetti, Denis Duboule, Immacolata Porreca, Nittoli, V., Fortunato, A. E., Fasano, G., Coppola, U., Gentile, A., Maiella, S., Langellotto, F., Porreca, I., De Paolo, R., Marino, R., Fiengo, M., Donizetti, A., Aniello, F., Kondo, T., Ristoratore, F., Canzoniero, L. M. T., Duboule, D., Wilson, S. W., and Sordino, P.
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0301 basic medicine ,Ce, cerebellum ,Mutant ,CNE, conserved non-coding elements ,OP, olfactory placode ,HM, hybridization mix ,0302 clinical medicine ,CaP, caudal primary motor neuron axons ,VLP, ventro-lateral-posterior ,Ey, eye ,Gene duplication ,No, notochord ,fss, fused-somites ,TSGD ,FB, forebrain ,Zebrafish ,OT, optic tectum ,CRM, cis-regulatory module ,Di, diencephalon ,CAMP, conserved ancestral microsyntenic pairs ,Signaling pathway ,Ace, acerebellar ,PSM, presomitic mesoderm ,CS, Corpuscle of Stannius ,General Medicine ,YE, yolk extension ,Cell biology ,ptc, patched ,Te, telencephalon ,030220 oncology & carcinogenesis ,PA, pharyngeal arches ,embryonic structures ,Shh, sonic hedgehog ,syu, sonic-you ,WISH, whole-mount in situ hybridization ,yot, you-too ,HB, hindbrain ,Hy, hypothalamus ,animal structures ,lcsh:QH426-470 ,SC, spinal cord ,MO, morpholino ,Development ,Biology ,Elfn1, Extracellular Leucine Rich Repeat And Fibronectin Type III Domain Containing 1 ,Synteny ,Uncx ,Article ,03 medical and health sciences ,Flh, floating head ,Mical, molecule interacting with CasL ,Th, thalamus ,Yo, yolk ,Genetics ,Gene ,Transcription factor ,Messenger RNA ,fungi ,biology.organism_classification ,cyc, cyclops ,FGF, fibroblast growth factor ,lcsh:Genetics ,smu, slow-muscle-omitted ,030104 developmental biology ,TSGD, teleost-specific genome duplication ,So, somites ,Homeobox ,AP, antero-posterior ,hpf, hours post fertilization ,WIHC, whole-mount immunohistochemistry - Abstract
The paired-type homeodomain transcription factor Uncx is involved in multiple processes of embryogenesis in vertebrates. Reasoning that zebrafish genes uncx4.1 and uncx are orthologs of mouse Uncx, we studied their genomic environment and developmental expression. Evolutionary analyses indicate the zebrafish uncx genes as being paralogs deriving from teleost-specific whole-genome duplication. Whole-mount in situ mRNA hybridization of uncx transcripts in zebrafish embryos reveals novel expression domains, confirms those previously known, and suggests sub-functionalization of paralogs. Using genetic mutants and pharmacological inhibitors, we investigate the role of signaling pathways on the expression of zebrafish uncx genes in developing somites. In identifying putative functional role(s) of zebrafish uncx genes, we hypothesized that they encode transcription factors that coordinate growth and innervation of somitic muscles., Highlights • The Uncx4.1 and Uncx genes derive from the teleost-specific whole-genome duplication. • Uncx genes are expressed during embryogenesis in unique and overlapping domains. • Uncx gene expression during somite differentiation is regulated by FGF signaling. • Synteny and expression profiles correlate Uncx genes with axon guidance.
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- 2019
9. Bisphenol A interferes with thyroid specific gene expression
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Emanuele Carchia, Erdentuya Ganbaatar, Daniela Gentilcore, Concetta Ambrosino, Massimo Mallardo, Immacolata Porreca, Mario De Felice, Francesca Rizzo, Gentilcore, D, Porreca, I, Rizzo, F, Ganbaatar, E, Carchia, E, Mallardo, Massimo, DE FELICE, Mario, and Ambrosino, C.
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Thyroid Hormones ,endocrine system ,medicine.medical_specialty ,Time Factors ,endocrine system diseases ,Thyroid Gland ,Thyroid ,Endocrine Disruptors ,Biology ,Toxicology ,Dose-Response Relationship ,Bisphenol A ,Phenols ,In vivo ,Transcription (biology) ,Internal medicine ,Gene expression ,medicine ,Animals ,Benzhydryl Compounds ,Thyroid specific gene transcription ,Transcription factor ,Zebrafish ,Regulation of gene expression ,Dose-Response Relationship, Drug ,urogenital system ,biology.organism_classification ,Rats ,Cell biology ,Endocrinology ,medicine.anatomical_structure ,Gene Expression Regulation ,Cell culture ,Pituitary Gland ,Cell line ,Environmental Pollutants ,Drug ,hormones, hormone substitutes, and hormone antagonists - Abstract
Bisphenol A (BPA) is an endocrine-disrupting chemical that leads to low-dose human exposure due to its ability to leach from chemically derived products, as polycarbonate plastics and epoxy resin. In addition to its known xeno-endocrine action, BPA exerts a wide range of metabolic effects. Despite the documented BPA exposure outcomes on synthesis of thyroid hormones, there are not any data available on its actions on the thyroid follicular cells, site of synthesis of the thyroid hormones. Recently, it has been shown that several environmental pollutants, as BPA, can exert a thyroid disrupting activity. In this study, we employed in vitro and in vivo (zebrafish) models to examine the effects of BPA in regulating the expression of genes involved in the thyroid hormone synthesis and of their transcriptional regulators at BPA doses as low as 10(-9)M, a dose that is environmentally pertinent and far below the one detected in infants plasma. In both systems we could detect an altered expression of the genes involved in thyroid hormones synthesis and of thyroid specific transcriptional factors in BPA dose and time dependent manner. Our results suggest that BPA exerts a direct effect on thyroid follicular cell. We show that these cells can "sense" very low amount of BPA. Thus they, potentially, represent an ideal in vitro system to develop assays to detect BPA and other pollutants with thyroid disrupting activity at level far below the ones considered to be environmental relevant. Moreover, this report may provide new insight into the mode of BPA-induced deregulation of physiological processes as well as on the extensively debated molecular pathways underlying its biological activities.
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- 2013
10. 'Stockpile' of Slight Transcriptomic Changes Determines the Indirect Genotoxicity of Low-Dose BPA in Thyroid Cells
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Concetta Ambrosino, Danila Cuomo, Immacolata Porreca, Massimo Mallardo, Mario De Felice, Luisa Ulloa Severino, Michele Ceccarelli, Angela Nebbioso, Elena Amendola, Lucia Altucci, Fulvio D'Angelo, Porreca, I, Ulloa Severino, L, D'Angelo, F, Cuomo, D, Ceccarelli, M, Altucci, L, Amendola, E, Nebbioso, A, Mallardo, M, De Felice, M, Ambrosino, C., Porreca, Immacolata, ULLOA SEVERINO, Luisa, D'Angelo, Fulvio, Cuomo, Danila, Ceccarelli, Michele, Altucci, Lucia, Amendola, Elena, Nebbioso, Angela, Mallardo, Massimo, De Felice, Mario, Ambrosino, Concetta, and Ulloa Severino, Luisa
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0301 basic medicine ,Genetics and Molecular Biology (all) ,Time Factors ,Light ,Benzhydryl Compounds ,Cell Line ,DNA Damage ,DNA Replication ,Dose-Response Relationship, Drug ,Gene Expression ,Humans ,Phenols ,Thyroid Gland ,Transcriptome ,Medicine (all) ,Biochemistry, Genetics and Molecular Biology (all) ,Agricultural and Biological Sciences (all) ,Microarrays ,DNA rapair ,lcsh:Medicine ,010501 environmental sciences ,medicine.disease_cause ,01 natural sciences ,Biochemistry ,Medicine and Health Sciences ,lcsh:Science ,Benzhydryl Compound ,Thyroid ,Multidisciplinary ,Physics ,Genomics ,Cell biology ,Nucleic acids ,Chemistry ,Bioassays and Physiological Analysis ,Toxicity ,Physical Sciences ,DNA microarray ,Drug ,Anatomy ,Transcriptome Analysis ,hormones, hormone substitutes, and hormone antagonists ,Human ,Research Article ,Ultraviolet radiation ,Programmed cell death ,endocrine system ,Pollutants ,Time Factor ,DNA repair ,DNA damage ,DNA recombination ,Endocrine System ,Biology ,Research and Analysis Methods ,Dose-Response Relationship ,03 medical and health sciences ,Electromagnetic radiation ,medicine ,Genetics ,Environmental Chemistry ,0105 earth and related environmental sciences ,Phenol ,Biology and life sciences ,urogenital system ,lcsh:R ,Ecology and Environmental Sciences ,Computational Biology ,DNA Repair Pathway ,DNA ,Genome Analysis ,Molecular biology ,030104 developmental biology ,lcsh:Q ,Ultraviolet C ,Genotoxicity - Abstract
Epidemiological and experimental data highlighted the thyroid-disrupting activity of bisphenol A (BPA). Although pivotal to identify the mechanisms of toxicity, direct low-dose BPA effects on thyrocytes have not been assessed. Here, we report the results of microarray experiments revealing that the transcriptome reacts dynamically to low-dose BPA exposure, adapting the changes in gene expression to the exposure duration. The response involves many genes, enriching specific pathways and biological functions mainly cell death/proliferation or DNA repair. Their expression is only slightly altered but, since they enrich specific pathways, this results in major effects as shown here for transcripts involved in the DNA repair pathway. Indeed, even though no phenotypic changes are induced by the treatment, we show that the exposure to BPA impairs the cell response to further stressors. We experimentally verify that prolonged exposure to low doses of BPA results in a delayed response to UV-C-induced DNA damage, due to impairment of p21-Tp53 axis, with the BPA-treated cells more prone to cell death and DNA damage accumulation. The present findings shed light on a possible mechanism by which BPA, not able to directly cause genetic damage at environmental dose, may exert an indirect genotoxic activity.
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- 2016
11. Zebrafish bcl2l is a survival factor in thyroid development
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Roberto Di Lauro, Henrik Fagman, Elena De Felice, Paolo Sordino, Immacolata Porreca, Porreca, I, De Felice, E, Fagman, H, DI LAURO, Roberto, and Sordino, P.
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endocrine system ,medicine.medical_specialty ,Programmed cell death ,endocrine system diseases ,Cell Survival ,Thyroid Gland ,bcl-X Protein ,Morphogenesis ,Biology ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,Gene expression ,medicine ,Animals ,Molecular Biology ,Transcription factor ,Zebrafish ,030304 developmental biology ,0303 health sciences ,Gene knockdown ,Thyroid ,Cell Biology ,Zebrafish Proteins ,biology.organism_classification ,Genes, bcl-2 ,Cell biology ,medicine.anatomical_structure ,Endocrinology ,030220 oncology & carcinogenesis ,Transcription Factors ,Developmental Biology ,Endocrine gland - Abstract
Regulated cell death, defined in morphological terms as apoptosis, is crucial for organ morphogenesis. While differentiation of the thyroid gland has been extensively studied, nothing is yet known about the survival mechanisms involved in the development of this endocrine gland. Using the zebrafish model system, we aim to understand whether genes belonging to the Bcl-2 family that control apoptosis are implicated in regulation of cell survival during thyroid development. Evidence of strong Bcl-2 gene expression in mouse thyroid precursors prompted us to investigate the functions played by its zebrafish homologs during thyroid development. We show that the bcl2-like (bcl2l) gene is expressed in the zebrafish thyroid primordium. Morpholino-mediated knockdown and mutant analyses revealed that bcl2l is crucial for thyroid cell survival and that this function is tightly modulated by the transcription factors pax2a, nk2.1a and hhex. Also, the bcl2l gene appears to control a caspase-3-dependent apoptotic mechanism during thyroid development. Thyroid precursor cells require an actively maintained survival mechanism to properly proceed through development. The bcl2l gene operates in the inhibition of cell death under direct regulation of a thyroid specific set of transcription factors. This is the first demonstration of an active mechanism to ensure survival of the thyroid primordium during morphogenesis. © 2012 Elsevier Inc.
- Published
- 2012
12. Evaluation of low doses BPA-induced perturbation of glycemia by toxicogenomics points to a primary role of pancreatic islets and to the mechanism of toxicity
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Immacolata Porreca, Concetta Ambrosino, Fulvio D'Angelo, M De Felice, Danila Cuomo, Massimo Mallardo, Emanuele Carchia, P. J. Almeida, Michele Ceccarelli, Carchia, E, Porreca, I, Almeida, Pj, D'Angelo, F, Cuomo, D, Ceccarelli, M, De Felice, M, Mallardo, M, and Ambrosino, C.
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Blood Glucose ,Male ,Cancer Research ,medicine.medical_specialty ,endocrine system ,Immunology ,BPA-induced ,glycemia ,toxicogenomics ,pancreatic ,toxicity ,Biology ,Toxicogenetics ,Islets of Langerhans ,Mice ,Cellular and Molecular Neuroscience ,Phenols ,In vivo ,Internal medicine ,medicine ,Animals ,Homeostasis ,Glucose homeostasis ,Benzhydryl Compounds ,geography ,geography.geographical_feature_category ,Dose-Response Relationship, Drug ,Pancreatic islets ,Cell Biology ,Islet ,Mice, Inbred C57BL ,Endocrinology ,medicine.anatomical_structure ,Toxicity ,Hepatocytes ,Original Article ,Toxicogenomics ,Ex vivo ,hormones, hormone substitutes, and hormone antagonists - Abstract
Epidemiologic and experimental studies have associated changes of blood glucose homeostasis to Bisphenol A (BPA) exposure. We took a toxicogenomic approach to investigate the mechanisms of low-dose (1 × 10−9M) BPA toxicity in ex vivo cultures of primary murine pancreatic islets and hepatocytes. Twenty-nine inhibited genes were identified in islets and none in exposed hepatocytes. Although their expression was slightly altered, their impaired cellular level, as a whole, resulted in specific phenotypic changes. Damage of mitochondrial function and metabolism, as predicted by bioinformatics analyses, was observed: BPA exposure led to a time-dependent decrease in mitochondrial membrane potential, to an increase of ROS cellular levels and, finally, to an induction of apoptosis, attributable to the bigger Bax/Bcl-2 ratio owing to activation of NF-κB pathway. Our data suggest a multifactorial mechanism for BPA toxicity in pancreatic islets with emphasis to mitochondria dysfunction and NF-κB activation. Finally, we assessed in vitro the viability of BPA-treated islets in stressing condition, as exposure to high glucose, evidencing a reduced ability of the exposed islets to respond to further damages. The result was confirmed in vivo evaluating the reduction of glycemia in hyperglycemic mice transplanted with control and BPA-treated pancreatic islets. The reported findings identify the pancreatic islet as the main target of BPA toxicity in impairing the glycemia. They suggest that the BPA exposure can weaken the response of the pancreatic islets to damages. The last observation could represent a broader concept whose consideration should lead to the development of experimental plans better reproducing the multiple exposure conditions.
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- 2015
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13. Localization of BDNF expression in the developing brain of zebrafish
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E. De Felice, E. Alleva, Antonino Germanà, Immacolata Porreca, P. de Girolamo, Concetta Ambrosino, Paolo Sordino, Emilia Ciriaco, De Felice, E, Porreca, I, Alleva, E, DE GIROLAMO, Paolo, Ambrosino, C, Ciriaco, E, Germanà, A, and Sordino, P.
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Histology ,EMX1 ,In situ hybridization ,Neurotrophic factors ,Animals ,RNA, Messenger ,Molecular Biology ,Zebrafish ,development ,Ecology, Evolution, Behavior and Systematics ,In Situ Hybridization ,Neurons ,Brain-derived neurotrophic factor ,biology ,Animal ,Reverse Transcriptase Polymerase Chain Reaction ,Brain-Derived Neurotrophic Factor ,Brain ,Gene Expression Regulation, Developmental ,Cell Biology ,Neuron ,biology.organism_classification ,central nervous system ,zebrafish ,eal-time quantitative polymerase chain reaction ,whole-mount in situ hybridizatio ,Molecular biology ,nervous system ,biology.protein ,whole-mount in situ hybridization ,Anatomy ,Tectum ,real-time quantitative polymerase chain reaction ,Neural development ,Neuroscience ,Research Article ,Developmental Biology ,Neurotrophin - Abstract
The brain-derived neurotrophic factor (BDNF) gene is expressed in differentiating and post-mitotic neurons of the zebrafish embryo, where it has been implicated in Huntington's disease. Little is known, however, about the full complement of neuronal cell types that express BDNF in this important vertebrate model. Here, we further explored the transcriptional profiles during the first week of development using real-time quantitative polymerase chain reaction (RT-qPCR) and whole-mount in situ hybridization (WISH). RT-qPCR results revealed a high level of maternal contribution followed by a steady increase of zygotic transcription, consistent with the notion of a prominent role of BDNF in neuronal maturation and maintenance. Based on WISH, we demonstrate for the first time that BDNF expression in the developing brain of zebrafish is structure specific. Anatomical criteria and co-staining with genetic markers (shh, pax2a, emx1, krox20, lhx2b and lhx9) visualized major topological domains of BDNF-positive cells in the pallium, hypothalamus, posterior tuberculum and optic tectum. Moreover, the relative timing of BDNF transcription in the eye and tectum may illustrate a mechanism for coordinated development of the retinotectal system. Taken together, our results are compatible with a local delivery and early role of BDNF in the developing brain of zebrafish, adding basic knowledge to the study of neurotrophin functions in neural development and disease.
- Published
- 2014
14. ONCOGENIC RAS BLOCKS THE cAMP PATHWAY AND DE-DIFFERENTIATES THYROID CELLS VIA AN IMPAIRMENT OF PAX8 TRANSCRIPTIONAL ACTIVITY
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Maria Giuseppina Baratta, Immacolata Porreca, Roberto Di Lauro, Baratta, M., Porreca, I., and DI LAURO, Roberto
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endocrine system ,Transcription, Genetic ,endocrine system diseases ,Oncogene Proteins ,Thyroid Gland ,Down-Regulation ,Biology ,Cell Line ,Mice ,PAX8 Transcription Factor ,Endocrinology ,Downregulation and upregulation ,Anti-apoptotic Ras signalling cascade ,Gene expression ,Cyclic AMP ,Animals ,Humans ,Paired Box Transcription Factors ,Molecular Biology ,Transcription factor ,Original Research ,Symporters ,Oncogene ,Receptors, Thyrotropin ,General Medicine ,Cell Dedifferentiation ,Cyclic AMP-Dependent Protein Kinases ,Molecular biology ,Rats ,Cell biology ,Kinetics ,Protein Transport ,Enhancer Elements, Genetic ,Genes, ras ,Organ Specificity ,cAMP-dependent pathway ,PAX8 ,Signal Transduction - Abstract
A deranged differentiation is often a landmark of transformed cells. We used a thyroid cell line expressing an inducible Ras oncoprotein in order to study the hierarchy of molecular events leading to suppression of thyroid-specific gene expression. We find that, upon Ras activation, there is an immediate global down-regulation of thyroid differentiation, which is associated with an inhibition of the cAMP signaling pathway. We demonstrate that an unusual negative cross talk between Ras oncogene and the cAMP pathway induces inactivation of the transcription factor Pax8 that we propose as a crucial event in Ras-induced dedifferentiation.
- Published
- 2009
15. An aptamer-mediated base editing platform for simultaneous knockin and multiple gene knockout for allogeneic CAR-T cells generation.
- Author
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Porreca I, Blassberg R, Harbottle J, Joubert B, Mielczarek O, Stombaugh J, Hemphill K, Sumner J, Pazeraitis D, Touza JL, Francescatto M, Firth M, Selmi T, Collantes JC, Strezoska Z, Taylor B, Jin S, Wiggins CM, van Brabant Smith A, and Lambourne JJ
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- Humans, Immunotherapy, Adoptive methods, Receptors, Chimeric Antigen genetics, Receptors, Chimeric Antigen metabolism, Gene Knock-In Techniques methods, Transgenes, Gene Editing methods, Gene Knockout Techniques, CRISPR-Cas Systems, Aptamers, Nucleotide genetics, T-Lymphocytes metabolism, T-Lymphocytes immunology
- Abstract
Gene editing technologies hold promise for enabling the next generation of adoptive cellular therapies. In conventional gene editing platforms that rely on nuclease activity, such as clustered regularly interspaced short palindromic repeats CRISPR-associated protein 9 (CRISPR-Cas9), allow efficient introduction of genetic modifications; however, these modifications occur via the generation of DNA double-strand breaks (DSBs) and can lead to unwanted genomic alterations and genotoxicity. Here, we apply a novel modular RNA aptamer-mediated Pin-point base editing platform to simultaneously introduce multiple gene knockouts and site-specific integration of a transgene in human primary T cells. We demonstrate high editing efficiency and purity at all target sites and significantly reduced frequency of chromosomal translocations compared with the conventional CRISPR-Cas9 system. Site-specific knockin of a chimeric antigen receptor and multiplex gene knockout are achieved within a single intervention and without the requirement for additional sequence-targeting components. The ability to perform complex genome editing efficiently and precisely highlights the potential of the Pin-point platform for application in a range of advanced cell therapies., Competing Interests: Declaration of interests M. Francescatto, M. Firth, J.L.T., D.P., J. Sumner, and B.T. are all current or past (while engaged in the research project) employees of AstraZeneca. I.P., R.B., J.H., B.J., O.M., J. Stombaugh, K.H., T.S., Z.S., C.W., A.v.B.S., and J.J.L. are current or past (while engaged in the research project) employees at Revvity. Revvity has an exclusive license from Rutgers University to certain base editing patents. Rutgers University and Horizon Discovery Limited have filed patent applications on this work., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2024
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16. Author Correction: Africa-specific human genetic variation near CHD1L associates with HIV-1 load.
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McLaren PJ, Porreca I, Iaconis G, Mok HP, Mukhopadhyay S, Karakoc E, Cristinelli S, Pomilla C, Bartha I, Thorball CW, Tough RH, Angelino P, Kiar CS, Carstensen T, Fatumo S, Porter T, Jarvis I, Skarnes WC, Bassett A, DeGorter MK, Sathya Moorthy MP, Tuff JF, Kim EY, Walter M, Simons LM, Bashirova A, Buchbinder S, Carrington M, Cossarizza A, De Luca A, Goedert JJ, Goldstein DB, Haas DW, Herbeck JT, Johnson EO, Kaleebu P, Kilembe W, Kirk GD, Kootstra NA, Kral AH, Lambotte O, Luo M, Mallal S, Martinez-Picado J, Meyer L, Miro JM, Moodley P, Motala AA, Mullins JI, Nam K, Obel N, Pirie F, Plummer FA, Poli G, Price MA, Rauch A, Theodorou I, Trkola A, Walker BD, Winkler CA, Zagury JF, Montgomery SB, Ciuffi A, Hultquist JF, Wolinsky SM, Dougan G, Lever AML, Gurdasani D, Groom H, Sandhu MS, and Fellay J
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- 2023
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17. Africa-specific human genetic variation near CHD1L associates with HIV-1 load.
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McLaren PJ, Porreca I, Iaconis G, Mok HP, Mukhopadhyay S, Karakoc E, Cristinelli S, Pomilla C, Bartha I, Thorball CW, Tough RH, Angelino P, Kiar CS, Carstensen T, Fatumo S, Porter T, Jarvis I, Skarnes WC, Bassett A, DeGorter MK, Sathya Moorthy MP, Tuff JF, Kim EY, Walter M, Simons LM, Bashirova A, Buchbinder S, Carrington M, Cossarizza A, De Luca A, Goedert JJ, Goldstein DB, Haas DW, Herbeck JT, Johnson EO, Kaleebu P, Kilembe W, Kirk GD, Kootstra NA, Kral AH, Lambotte O, Luo M, Mallal S, Martinez-Picado J, Meyer L, Miro JM, Moodley P, Motala AA, Mullins JI, Nam K, Obel N, Pirie F, Plummer FA, Poli G, Price MA, Rauch A, Theodorou I, Trkola A, Walker BD, Winkler CA, Zagury JF, Montgomery SB, Ciuffi A, Hultquist JF, Wolinsky SM, Dougan G, Lever AML, Gurdasani D, Groom H, Sandhu MS, and Fellay J
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- Humans, Cell Line, Africa, Chromosomes, Human, Pair 1 genetics, Alleles, RNA, Long Noncoding genetics, Virus Replication, DNA Helicases genetics, DNA Helicases metabolism, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Genetic Variation, HIV Infections genetics, HIV-1 growth & development, HIV-1 physiology, Viral Load genetics
- Abstract
HIV-1 remains a global health crisis
1 , highlighting the need to identify new targets for therapies. Here, given the disproportionate HIV-1 burden and marked human genome diversity in Africa2 , we assessed the genetic determinants of control of set-point viral load in 3,879 people of African ancestries living with HIV-1 participating in the international collaboration for the genomics of HIV3 . We identify a previously undescribed association signal on chromosome 1 where the peak variant associates with an approximately 0.3 log10 -transformed copies per ml lower set-point viral load per minor allele copy and is specific to populations of African descent. The top associated variant is intergenic and lies between a long intergenic non-coding RNA (LINC00624) and the coding gene CHD1L, which encodes a helicase that is involved in DNA repair4 . Infection assays in iPS cell-derived macrophages and other immortalized cell lines showed increased HIV-1 replication in CHD1L-knockdown and CHD1L-knockout cells. We provide evidence from population genetic studies that Africa-specific genetic variation near CHD1L associates with HIV replication in vivo. Although experimental studies suggest that CHD1L is able to limit HIV infection in some cell types in vitro, further investigation is required to understand the mechanisms underlying our observations, including any potential indirect effects of CHD1L on HIV spread in vivo that our cell-based assays cannot recapitulate., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2023
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18. Chronic Exposure to Chlorpyrifos Damages Thyroid Activity and Imbalances Hepatic Thyroid Hormones Signaling and Glucose Metabolism: Dependency of T3-FOXO1 Axis by Hyperglycemia.
- Author
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Peluso T, Nittoli V, Reale C, Porreca I, Russo F, Roberto L, Giacco A, Silvestri E, Mallardo M, De Felice M, and Ambrosino C
- Subjects
- Animals, Male, Mice, Glucose metabolism, Liver metabolism, Thyroid Gland metabolism, Thyroid Hormones metabolism, Iodothyronine Deiodinase Type II, Chlorpyrifos metabolism, Hyperglycemia chemically induced, Hyperglycemia metabolism
- Abstract
Early life exposure to Endocrine Disruptor Chemicals (EDCs), such as the organophosphate pesticide Chlorpyrifos (CPF), affects the thyroid activity and dependent process, including the glucose metabolism. The damage of thyroid hormones (THs) as a mechanism of action of CPF is underestimated because the studies rarely consider that TH levels and signaling are customized peripherally. Here, we investigated the impairment of metabolism/signaling of THs and lipid/glucose metabolism in the livers of 6-month-old mice, developmentally and lifelong exposed to 0.1, 1, and 10 mg/kg/die CPF (F1) and their offspring similarly exposed (F2), analyzing the levels of transcripts of the enzymes involved in the metabolism of T3 ( Dio1 ), lipids ( Fasn , Acc1 ), and glucose ( G6pase , Pck1 ). Both processes were altered only in F2 males, affected by hypothyroidism and by a systemic hyperglycemia linked to the activation of gluconeogenesis in mice exposed to 1 and 10 mg/kg/die CPF. Interestingly, we observed an increase in active FOXO1 protein due to a decrease in AKT phosphorylation, despite insulin signaling activation. Experiments in vitro revealed that chronic exposure to CPF affected glucose metabolism via the direct modulation of FOXO1 activity and T3 levels in hepatic cells. In conclusion, we described different sex and intergenerational effects of CPF exposure on the hepatic homeostasis of THs, their signaling, and, finally, glucose metabolism. The data points to FOXO1-T3-glucose signaling as a target of CPF in liver.
- Published
- 2023
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19. The p66Shc Protein Mediates Insulin Resistance and Secretory Dysfunction in Pancreatic β-Cells Under Lipotoxic Conditions.
- Author
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Biondi G, Marrano N, Dipaola L, Borrelli A, Rella M, D'Oria R, Genchi VA, Caccioppoli C, Porreca I, Cignarelli A, Perrini S, Marchetti P, Vincenti L, Laviola L, Giorgino F, and Natalicchio A
- Subjects
- Animals, Apoptosis, C-Peptide metabolism, Humans, Insulin metabolism, Mice, Palmitates metabolism, Palmitates pharmacology, Signal Transduction, Src Homology 2 Domain-Containing, Transforming Protein 1 genetics, Insulin Resistance, Insulin-Secreting Cells metabolism, Src Homology 2 Domain-Containing, Transforming Protein 1 metabolism
- Abstract
We evaluated the role of the p66Shc redox adaptor protein in pancreatic β-cell insulin resistance that develops under lipotoxic conditions and with excess body fat. Prolonged exposure to palmitate in vitro or the presence of overweight/obesity augmented p66Shc expression levels and caused an impaired ability of exogenous insulin to increase cellular insulin content and secreted C-peptide levels in INS-1E cells and human and murine islets. In INS-1E cells, p66Shc knockdown resulted in enhanced insulin-induced augmentation of insulin content and C-peptide secretion and prevented the ability of palmitate to impair these effects of insulin. Conversely, p66Shc overexpression impaired insulin-induced augmentation of insulin content and C-peptide secretion in both the absence and presence of palmitate. Under lipotoxic condition, the effects of p66Shc are mediated by a p53-induced increase in p66Shc protein levels and JNK-induced p66Shc phosphorylation at Ser36 and appear to involve the phosphorylation of the ribosomal protein S6 kinase at Thr389 and of insulin receptor substrate 1 at Ser307, resulting in the inhibition of insulin-stimulated protein kinase B phosphorylation at Ser473. Thus, the p66Shc protein mediates the impaired β-cell function and insulin resistance induced by saturated fatty acids and excess body fat., (© 2022 by the American Diabetes Association.)
- Published
- 2022
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20. ZFP36L2 Role in Thyroid Functionality.
- Author
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Albano F, Tucci V, Blackshear PJ, Reale C, Roberto L, Russo F, Marotta P, Porreca I, Colella M, Mallardo M, de Felice M, and Ambrosino C
- Subjects
- Animals, Apoptosis physiology, Cell Line, Cell Survival, Female, Gene Deletion, Gene Expression Regulation, Mice, Inbred C57BL, Mice, Mutant Strains, PAX8 Transcription Factor genetics, Rats, Receptor, Notch1 metabolism, Thyroid Gland cytology, Thyroid Gland drug effects, Thyrotropin pharmacology, Tristetraprolin genetics, Mice, Thyroid Gland physiology, Tristetraprolin physiology
- Abstract
Thyroid hormone levels are usually genetically determined. Thyrocytes produce a unique set of enzymes that are dedicated to thyroid hormone synthesis. While thyroid transcriptional regulation is well-characterized, post-transcriptional mechanisms have been less investigated. Here, we describe the involvement of ZFP36L2, a protein that stimulates degradation of target mRNAs, in thyroid development and function, by in vivo and in vitro gene targeting in thyrocytes. Thyroid-specific Zfp36l2
-/- females were hypothyroid, with reduced levels of circulating free Thyroxine (cfT4) and Triiodothyronine (cfT3). Their hypothyroidism was due to dyshormonogenesis, already evident one week after weaning, while thyroid development appeared normal. We observed decreases in several thyroid-specific transcripts and proteins, such as Nis and its transcriptional regulators ( Pax8 and Nkx2 .1), and increased apoptosis in Zfp36l2-/- thyroids. Nis , Pax8, and Nkx2 .1 mRNAs were also reduced in Zfp36l2 knock-out thyrocytes in vitro (L2KO), in which we confirmed the increased apoptosis. Finally, in L2KO cells, we showed an altered response to TSH stimulation regarding both thyroid-specific gene expression and cell proliferation and survival. This result was supported by increases in P21/WAF1 and p-P38MAPK levels. Mechanistically, we confirmed Notch1 as a target of ZFP36L2 in the thyroid since its levels were increased in both in vitro and in vivo models. In both models, the levels of Id4 mRNA, a potential inhibitor of Pax8 activity, were increased. Overall, the data indicate that the regulation of mRNA stability by ZFP36L2 is a mechanism that controls the function and survival of thyrocytes.- Published
- 2021
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21. Multi Species Analyses Reveal Testicular T3 Metabolism and Signalling as a Target of Environmental Pesticides.
- Author
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Nittoli V, Colella M, Porciello A, Reale C, Roberto L, Russo F, Russo NA, Porreca I, De Felice M, Mallardo M, and Ambrosino C
- Subjects
- Animals, Cell Differentiation drug effects, Germ Cells drug effects, Germ Cells metabolism, Male, Mice, Mice, Inbred C57BL, Reproduction drug effects, Seminiferous Tubules drug effects, Seminiferous Tubules metabolism, Spermatogenesis drug effects, Testis metabolism, Thyroid Hormones metabolism, Zebrafish metabolism, Pesticides pharmacology, Signal Transduction drug effects, Testis diagnostic imaging
- Abstract
Thyroid hormones (THs) regulate many biological processes in vertebrates, including reproduction. Testicular somatic and germ cells are equipped with the arrays of enzymes (deiodinases), transporters, and receptors necessary to locally maintain the optimal level of THs and their signalling, needed for their functions and spermatogenesis. Pesticides, as chlorpyrifos (CPF) and ethylene thiourea (ETU), impair the function of thyroid and testis, affecting male fertility. However, their ability to disarrange testicular T3 (t-T3) metabolism and signalling is poorly considered. Here, a multi-species analysis involving zebrafish and mouse suggests the damage of t-T3 metabolism and signalling as a mechanism of gonadic toxicity of low-doses CPF and ETU. Indeed, the developmental exposure to both compounds reduces Dio2 transcript in both models, as well as in ex-vivo cultures of murine seminiferous tubules, and it is linked to alteration of steroidogenesis and germ cell differentiation. A major impact on spermatogonia was confirmed molecularly by the expression of their markers and morphologically evidenced in zebrafish. The results reveal that in the adopted models, exposure to both pesticides alters the t-T3 metabolism and signalling, affecting the reproductive capability. Our data, together with previous reports suggest zebrafish as an evaluable model in assessing the action of compounds impairing locally T3 signalling.
- Published
- 2021
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22. Peripheral T3 signaling is the target of pesticides in zebrafish larvae and adult liver.
- Author
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Colella M, Nittoli V, Porciello A, Porreca I, Reale C, Russo F, Russo NA, Roberto L, Albano F, De Felice M, Mallardo M, and Ambrosino C
- Subjects
- Animals, Chlorpyrifos administration & dosage, Chlorpyrifos adverse effects, Endocrine Disruptors, Ethylenethiourea administration & dosage, Ethylenethiourea adverse effects, Female, Forkhead Box Protein O3 metabolism, Larva drug effects, Liver drug effects, Male, PPAR alpha metabolism, Signal Transduction physiology, Thyroid Gland growth & development, Thyroid Gland metabolism, Thyroxine metabolism, Zebrafish growth & development, Larva metabolism, Liver metabolism, Pesticides adverse effects, Signal Transduction drug effects, Triiodothyronine metabolism, Zebrafish metabolism
- Abstract
The intra-tissue levels of thyroid hormones (THs) regulate organ functions. Environmental factors can impair these levels by damaging the thyroid gland and/or peripheral TH metabolism. We investigated the effects of embryonic and/or long-life exposure to low-dose pesticides, ethylene thiourea (ETU), chlorpyrifos (CPF) and both combined on intra-tissue T4/T3 metabolism/signaling in zebrafish at different life stages. Hypothyroidism was evident in exposed larvae that showed reduced number of follicles and induced tshb mRNAs. Despite that, we found an increase in free T4 (fT4) and free T3 (fT3) levels/signaling that was confirmed by transcriptional regulation of TH metabolic enzymes (deiodinases) and T3-regulated mRNAs (cpt1, igfbp1a). Second-generation larvae showed that thyroid and TH signaling was affected even when not directly exposed, suggesting the role of parental exposure. In adult zebrafish, we found that sex-dependent damage of hepatic T3 level/signaling was associated with liver steatosis, which was more pronounced in females, with sex-dependent alteration of transcripts codifying the key enzymes involved in 'de novo lipogenesis' and β-oxidation. We found impaired activation of liver T3 and PPARα/Foxo3a pathways whose deregulation was already involved in mammalian liver steatosis. The data emphasizes that the intra-tissue imbalance of the T3 level is due to thyroid endocrine disruptors (THDC) and suggests that the effect of a slight modification in T3 signaling might be amplified by its direct regulation or crosstalk with PPARα/Foxo3a pathways. Because T3 levels define the hypothyroid/hyperthyroid status of each organ, our findings might explain the pleiotropic and site-dependent effects of pesticides.
- Published
- 2020
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23. Loss of IL-10 signaling in macrophages limits bacterial killing driven by prostaglandin E2.
- Author
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Mukhopadhyay S, Heinz E, Porreca I, Alasoo K, Yeung A, Yang HT, Schwerd T, Forbester JL, Hale C, Agu CA, Choi YH, Rodrigues J, Capitani M, Jostins-Dean L, Thomas DC, Travis S, Gaffney D, Skarnes WC, Thomson N, Uhlig HH, Dougan G, and Powrie F
- Subjects
- Cell Differentiation genetics, Cells, Cultured, Dinoprostone antagonists & inhibitors, Female, Gene Knockout Techniques, Humans, Induced Pluripotent Stem Cells metabolism, Inflammation chemically induced, Inflammation metabolism, Inflammatory Bowel Diseases pathology, Interleukin-10 Receptor alpha Subunit genetics, Interleukin-10 Receptor beta Subunit genetics, Lipopolysaccharides pharmacology, Macrophage Activation drug effects, Macrophage Activation genetics, Macrophages drug effects, Mutation, Phosphorylation genetics, STAT3 Transcription Factor metabolism, Signal Transduction drug effects, Dinoprostone metabolism, Inflammatory Bowel Diseases metabolism, Interleukin-10 metabolism, Interleukin-10 Receptor beta Subunit metabolism, Macrophages metabolism, Salmonella typhimurium metabolism, Signal Transduction genetics
- Abstract
Loss of IL-10 signaling in macrophages (Mφs) leads to inflammatory bowel disease (IBD). Induced pluripotent stem cells (iPSCs) were generated from an infantile-onset IBD patient lacking a functional IL10RB gene. Mφs differentiated from IL-10RB-/- iPSCs lacked IL-10RB mRNA expression, were unable to phosphorylate STAT3, and failed to reduce LPS induced inflammatory cytokines in the presence of exogenous IL-10. IL-10RB-/- Mφs exhibited a striking defect in their ability to kill Salmonella enterica serovar Typhimurium, which was rescuable after experimentally introducing functional copies of the IL10RB gene. Genes involved in synthesis and receptor pathways for eicosanoid prostaglandin E2 (PGE2) were more highly induced in IL-10RB-/- Mφs, and these Mφs produced higher amounts of PGE2 after LPS stimulation compared with controls. Furthermore, pharmacological inhibition of PGE2 synthesis and PGE2 receptor blockade enhanced bacterial killing in Mφs. These results identify a regulatory interaction between IL-10 and PGE2, dysregulation of which may drive aberrant Mφ activation and impaired host defense contributing to IBD pathogenesis., (© 2019 Mukhopadhyay et al.)
- Published
- 2020
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24. Characterization of paralogous uncx transcription factor encoding genes in zebrafish.
- Author
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Nittoli V, Fortunato AE, Fasano G, Coppola U, Gentile A, Maiella S, Langellotto F, Porreca I, De Paolo R, Marino R, Fiengo M, Donizetti A, Aniello F, Kondo T, Ristoratore F, Canzoniero LMT, Duboule D, Wilson SW, and Sordino P
- Abstract
The paired-type homeodomain transcription factor Uncx is involved in multiple processes of embryogenesis in vertebrates. Reasoning that zebrafish genes uncx4.1 and uncx are orthologs of mouse Uncx , we studied their genomic environment and developmental expression. Evolutionary analyses indicate the zebrafish uncx genes as being paralogs deriving from teleost-specific whole-genome duplication. Whole-mount in situ mRNA hybridization of uncx transcripts in zebrafish embryos reveals novel expression domains, confirms those previously known, and suggests sub-functionalization of paralogs. Using genetic mutants and pharmacological inhibitors, we investigate the role of signaling pathways on the expression of zebrafish uncx genes in developing somites. In identifying putative functional role(s) of zebrafish uncx genes, we hypothesized that they encode transcription factors that coordinate growth and innervation of somitic muscles.
- Published
- 2019
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25. A Toxicogenomic Approach Reveals a Novel Gene Regulatory Network Active in In Vitro and In Vivo Models of Thyroid Carcinogenesis.
- Author
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Reale C, Russo F, Credendino SC, Cuomo D, De Vita G, Mallardo M, Pennino F, Porreca I, Triassi M, De Felice M, and Ambrosino C
- Subjects
- Animals, Carcinogenesis, Carcinogens toxicity, Cell Line, Chlorpyrifos toxicity, Female, Maternal-Fetal Exchange, Mice, Mitogen-Activated Protein Kinases metabolism, Polychlorinated Dibenzodioxins toxicity, Pregnancy, Rats, Receptors, Cell Surface genetics, Thyroid Epithelial Cells drug effects, Thyroid Epithelial Cells metabolism, Transcription Factors genetics, Tristetraprolin genetics, Gene Regulatory Networks, Thyroid Neoplasms genetics
- Abstract
Epidemiological and experimental studies emphasize the link between environmental chemicals exposure and thyroid cancer. However, this association is strongly debated and the mechanisms of action of environmental thyroid carcinogens still need to be identified. The analysis of in vitro transcriptomic data developed to investigate the effects of chlorpyrifos on immortalized thyrocytes highlighted the impaired expression of genes involved in endodermal carcinogenesis. This endodermal carcinogenic gene-network (ECGN, including Zfp36l2 , Dmbt1 , Ddit4 ), was validated in cellular and mouse models of thyroid carcinogenesis, characterized by the constitutive activation of the mitogen-activated protein kinase (MAPK) pathway and in immortalized thyrocytes exposed to tetrachlorodibenzo- p -dioxin (TCDD) and chlorpyrifos (CPF). The mRNA levels of Zfp36l2 , Dmbt1 and Ddit4 were increased in models characterized by MAPK activation or following TCDD exposure, whereas they were inhibited by CPF exposure. Overall, the ECGN transcripts identify a novel gene-regulatory network associated with thyroid carcinogenesis promoted by genetic mutation or by environmental carcinogens. The latter have opposite effects on the modulation of the ECGN transcripts according to their mechanisms of action in promoting carcinogenesis. Therefore, the analyses of ECGN might be helpful in discriminating compounds that promote cellular survival associated or not to proliferation of thyrocytes.
- Published
- 2019
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26. Transcriptional landscape of mouse-aged ovaries reveals a unique set of non-coding RNAs associated with physiological and environmental ovarian dysfunctions.
- Author
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Cuomo D, Porreca I, Ceccarelli M, Threadgill DW, Barrington WT, Petriella A, D'Angelo F, Cobellis G, De Stefano F, D'Agostino MN, De Felice M, Mallardo M, and Ambrosino C
- Abstract
The progressive and physiological decline in ovarian function depends on the rate of follicular loss by atresia, contributing to the reduction in ovarian reserve. Genetics and environmental factors play important roles in ovarian senescence and in the onset of ovarian dysfunctions such as diminished ovarian reserve. A better understanding of the mechanisms underlying ovarian aging and their regulation by genetic and environmental factors is needed to evaluate ovarian reserve and to predict fertility potential by identification of more accurate and less invasive markers. We report transcriptomic data (i) implicating novel (e.g. EIF2 signalling) and well-known pathways (e.g. TGFβ signalling), and (ii) defining a unique set of non-coding RNA (ncRNA), both associated with ovarian function. The latter includes miRNAs (e.g. Mir143 and Mir145 ), snoRNAs (e.g. Snord16a and Snora34 ), and one lncRNA ( Gas5 ), which are differentially expressed in middle-aged ovaries (12 months) vs young-aged (3 months) from CD1 mice. Experimental analysis confirms that ovary lifespan varies across genetic backgrounds in mice and, genetics influences the response to environmental perturbations such as diet. Moreover, the identified ncRNAs were verified in a model of reproductive dysfunction promoted by the environmental toxicant ethylenthiourea. We also report the increase of miRNA143 and miRNA145 in follicular fluid of women with diminished ovarian reserve. Their levels inversely correlate with the hormonal profile and with the number of the oocytes recruited upon hormonal stimulation. Overall, we report a transcriptomic signature for ovarian dysfunction in vivo that provides a valuable resource for translational research in human reproductive aging., Competing Interests: The authors declare that they have no conflict of interest.
- Published
- 2018
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27. Genetic background and window of exposure contribute to thyroid dysfunction promoted by low-dose exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin in mice.
- Author
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Reale C, Porreca I, Russo F, Marotta M, Roberto L, Russo NA, Carchia E, Mallardo M, De Felice M, and Ambrosino C
- Subjects
- Animals, Cell Line, Dose-Response Relationship, Drug, Female, Haploinsufficiency, Hypothyroidism chemically induced, Hypothyroidism genetics, Hypothyroidism metabolism, Hypothyroidism physiopathology, Male, Mice, NF-KappaB Inhibitor alpha metabolism, PAX8 Transcription Factor genetics, Phenotype, Sex Characteristics, Signal Transduction drug effects, Thyroid Gland cytology, Thyroid Nuclear Factor 1 genetics, Thyroid Nuclear Factor 1 metabolism, Time Factors, Transcription Factor RelA metabolism, Tumor Suppressor Protein p53 metabolism, Polychlorinated Dibenzodioxins toxicity, Thyroid Gland drug effects, Thyroid Gland physiopathology
- Abstract
Genetic and environmental factors contribute to thyroid diseases. Although still debated, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is thought to induce thyroid dysfunction in humans and rodents. The data here reported point out the contribution of the exposure window and genetic background in mediating the low-dose TCDD effects on thyroid. Indeed, early (from E0.5 to PND30) and low-dose (0,001 μg/kg/day) TCDD exposure reduced the circulating fT4 and altered the expression of thyroid specific transcripts. The role of genetic components was estimated monitoring the same markers in Pax8
+/- and Nkx2-1+/- mice, susceptible to thyroid dysfunction, exposed to 0, 1 μg/kg/day TCDD from E15.5 to PND60. Haploinsufficiency of either Pax8 or Nkx2-1 genes exacerbated the effects of the exposure impairing the thyroid enriched mRNAs in sex dependent manner. Such effect was mediated by mechanisms involving the Nkx2-1/p53/p65/IĸBα pathway in vitro and in vivo. Foetal exposure to TCDD impaired both thyroid function and genes expression while thyroid development and differentiation did not appear significantly affected. In mouse, stronger effects were related to earlier exposure or specific genetic background such as either Pax8 or Nkx2-1 haploinsufficiency, both associated to hypothyroidism in humans. Furthermore, our data underline that long exposure time are needed to model in vitro and in vivo results.- Published
- 2018
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28. Bisphenol a and mesenchymal stem cells: Recent insights.
- Author
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Nunes HC, Scarano WR, Deffune E, Felisbino SL, Porreca I, and Delella FK
- Subjects
- Adipogenesis drug effects, Adipose Tissue cytology, Adipose Tissue drug effects, Animals, Humans, Obesity chemically induced, Osteogenesis drug effects, Benzhydryl Compounds adverse effects, Endocrine Disruptors adverse effects, Mesenchymal Stem Cells cytology, Mesenchymal Stem Cells drug effects, Phenols adverse effects
- Published
- 2018
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29. Carcinogenic risk and Bisphenol A exposure: A focus on molecular aspects in endoderm derived glands.
- Author
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Cuomo D, Porreca I, Cobellis G, Tarallo R, Nassa G, Falco G, Nardone A, Rizzo F, Mallardo M, and Ambrosino C
- Subjects
- Animals, Humans, Neoplasms genetics, Neoplasms pathology, Risk Factors, Benzhydryl Compounds toxicity, Carcinogenesis genetics, Carcinogenesis pathology, Endoderm pathology, Environmental Exposure analysis, Phenols toxicity
- Abstract
Epidemiological and experimental evidence associates the exposure to Bisphenol A with the increase of cancer risk in several organs, including prostate. BPA targets different pathways involved in carcinogenicity including the Nuclear Receptors (i.e. estrogen and androgen receptors), stress regulated proteins and, finally, epigenetic changes. Here, we analyse BPA-dependent carcinogenesis in endoderm-derived glands, thyroid, liver, pancreas and prostate focusing on cell signalling, DNA damage repair pathways and epigenetic modifications. Mainly, we gather molecular data evidencing harmful effects at doses relevant for human risk (low-doses). Since few molecular data are available, above all for the pancreas, we analysed transcriptomic data generated in our laboratory to suggest possible mechanisms of BPA carcinogenicity in endoderm-derived glands, discussing the role of nuclear receptors and stress/NF-kB pathways. We evidence that an in vitro toxicogenomic approach might suggest mechanisms of toxicity applicable to cells having the same developmental origin. Although we cannot draw firm conclusions, published data summarized in this review suggest that exposure to BPA, primarily during the developmental stages, represents a risk for carcinogenesis of endoderm-derived glands., (Copyright © 2017. Published by Elsevier B.V.)
- Published
- 2017
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30. Specific Effects of Chronic Dietary Exposure to Chlorpyrifos on Brain Gene Expression-A Mouse Study.
- Author
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Pallotta MM, Ronca R, Carotenuto R, Porreca I, Turano M, Ambrosino C, and Capriglione T
- Subjects
- Animals, Brain drug effects, Brain pathology, Dietary Exposure adverse effects, Female, Gene Expression Regulation drug effects, Humans, Insecticides toxicity, Mice, Parkinson Disease, Secondary chemically induced, Parkinson Disease, Secondary pathology, Protein Biosynthesis drug effects, Brain metabolism, Chlorpyrifos toxicity, Maternal Exposure adverse effects, Parkinson Disease, Secondary genetics
- Abstract
chlorpyrifos (CPF) is an organophosphate insecticide used to control pests on a variety of food and feed crops. In mammals, maternal exposure to CPF has been reported to induce cerebral cortex thinning, alteration of long-term brain cognitive function, and Parkinson-like symptoms, but the mechanisms of these processes are not fully understood. In this study, we aimed to gain a deeper understanding of the alterations induced in the brains of mice chronically exposed to CPF by dietary intake. For our purpose, we analysed F1 offspring (sacrificed at 3 and 8 months) of Mus musculus , treated in utero and postnatally with 3 different doses of CPF (0.1-1-10 mg/kg/day). Using RT² Profiler PCR Arrays, we evaluated the alterations in the expression of 84 genes associated with neurodegenerative diseases. In the brains of exposed mice, we evidenced a clear dose-response relationship for AChE inhibition and alterations of gene expression. Some of the genes that were steadily down-regulated, such as Pink1 , Park 2 , Sv2b , Gabbr2 , Sept5 and Atxn2 , were directly related to Parkinson's onset. Our experimental results shed light on the possibility that long-term CPF exposure may exert membrane signalling alterations which make brain cells more susceptible to develop neurodegenerative diseases.
- Published
- 2017
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31. The Myokine Irisin Is Released in Response to Saturated Fatty Acids and Promotes Pancreatic β-Cell Survival and Insulin Secretion.
- Author
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Natalicchio A, Marrano N, Biondi G, Spagnuolo R, Labarbuta R, Porreca I, Cignarelli A, Bugliani M, Marchetti P, Perrini S, Laviola L, and Giorgino F
- Subjects
- Animals, Apoptosis drug effects, Cell Proliferation drug effects, Fibronectins genetics, Fibronectins pharmacology, Gene Expression Regulation physiology, Glucose, Humans, Insulin Secretion, Islets of Langerhans cytology, Male, Mice, Mice, Inbred C57BL, Muscle Fibers, Skeletal drug effects, Palmitates toxicity, Random Allocation, Rats, Recombinant Proteins pharmacology, Cell Survival physiology, Fatty Acids pharmacology, Fibronectins metabolism, Insulin metabolism, Insulin-Secreting Cells physiology, Islets of Langerhans drug effects
- Abstract
This study explored the role of irisin as a new pancreatic β-cell secretagogue and survival factor and its potential role in the communication between skeletal muscle and pancreatic β-cells under lipotoxic conditions. Recombinant irisin stimulated insulin biosynthesis and glucose-stimulated insulin secretion (GSIS) in a PKA-dependent manner and prevented saturated fatty acid-induced apoptosis in human and rat pancreatic β-cells, as well as in human and murine pancreatic islets, via AKT/BCL2 signaling. Treatment of myotubes with 0.5 mmol/L palmitate for 4 h, but not with oleate, promoted an increase in irisin release in the culture medium. Moreover, increased serum levels of irisin were observed in mice fed with a high-fat diet. Mouse serum rich in irisin and the conditioned medium from myotubes exposed to palmitate for 4 h significantly reduced apoptosis of murine pancreatic islets and insulin-secreting INS-1E cells, respectively, and this was abrogated in the presence of an irisin-neutralizing antibody. Finally, in vivo administration of irisin improved GSIS and increased β-cell proliferation. In conclusion, irisin can promote β-cell survival and enhance GSIS and may thus participate in the communication between skeletal muscle and β-cells under conditions of excess saturated fatty acids., (© 2017 by the American Diabetes Association.)
- Published
- 2017
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32. Pesticide toxicogenomics across scales: in vitro transcriptome predicts mechanisms and outcomes of exposure in vivo.
- Author
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Porreca I, D'Angelo F, De Franceschi L, Mattè A, Ceccarelli M, Iolascon A, Zamò A, Russo F, Ravo M, Tarallo R, Scarfò M, Weisz A, De Felice M, Mallardo M, and Ambrosino C
- Subjects
- Animals, Cells, Cultured, Chlorpyrifos toxicity, Ethylenethiourea toxicity, Female, Gene Expression Profiling, Hematopoiesis drug effects, Humans, Male, Mice, Rats, Thyroid Gland cytology, Thyroid Gland drug effects, Thyroid Gland metabolism, Toxicity Tests methods, Pesticides toxicity, Transcriptome drug effects
- Abstract
In vitro Omics analysis (i.e. transcriptome) is suggested to predict in vivo toxicity and adverse effects in humans, although the causal link between high-throughput data and effects in vivo is not easily established. Indeed, the chemical-organism interaction can involve processes, such as adaptation, not established in cell cultures. Starting from this consideration we investigate the transcriptomic response of immortalized thyrocytes to ethylenthiourea and chlorpyrifos. In vitro data revealed specific and common genes/mechanisms of toxicity, controlling the proliferation/survival of the thyrocytes and unrelated hematopoietic cell lineages. These results were phenotypically confirmed in vivo by the reduction of circulating T4 hormone and the development of pancytopenia after long exposure. Our data imply that in vitro toxicogenomics is a powerful tool in predicting adverse effects in vivo, experimentally confirming the vision described as Tox21c (Toxicity Testing in the 21st century) although not fully recapitulating the biocomplexity of a living animal.
- Published
- 2016
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33. "Stockpile" of Slight Transcriptomic Changes Determines the Indirect Genotoxicity of Low-Dose BPA in Thyroid Cells.
- Author
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Porreca I, Ulloa Severino L, D'Angelo F, Cuomo D, Ceccarelli M, Altucci L, Amendola E, Nebbioso A, Mallardo M, De Felice M, and Ambrosino C
- Subjects
- Benzhydryl Compounds administration & dosage, Cell Line, DNA Damage, DNA Replication genetics, Dose-Response Relationship, Drug, Gene Expression drug effects, Humans, Phenols administration & dosage, Thyroid Gland metabolism, Time Factors, Benzhydryl Compounds toxicity, Phenols toxicity, Thyroid Gland drug effects, Transcriptome
- Abstract
Epidemiological and experimental data highlighted the thyroid-disrupting activity of bisphenol A (BPA). Although pivotal to identify the mechanisms of toxicity, direct low-dose BPA effects on thyrocytes have not been assessed. Here, we report the results of microarray experiments revealing that the transcriptome reacts dynamically to low-dose BPA exposure, adapting the changes in gene expression to the exposure duration. The response involves many genes, enriching specific pathways and biological functions mainly cell death/proliferation or DNA repair. Their expression is only slightly altered but, since they enrich specific pathways, this results in major effects as shown here for transcripts involved in the DNA repair pathway. Indeed, even though no phenotypic changes are induced by the treatment, we show that the exposure to BPA impairs the cell response to further stressors. We experimentally verify that prolonged exposure to low doses of BPA results in a delayed response to UV-C-induced DNA damage, due to impairment of p21-Tp53 axis, with the BPA-treated cells more prone to cell death and DNA damage accumulation. The present findings shed light on a possible mechanism by which BPA, not able to directly cause genetic damage at environmental dose, may exert an indirect genotoxic activity.
- Published
- 2016
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34. The "busy life" of unliganded estrogen receptors.
- Author
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Stellato C, Porreca I, Cuomo D, Tarallo R, Nassa G, and Ambrosino C
- Subjects
- Animals, Estrogens physiology, Gene Expression Regulation, Humans, Ligands, Proteome genetics, Proteome metabolism, Proteomics, Signal Transduction, Receptors, Estrogen physiology
- Abstract
Understanding of the role of estrogen receptors (ERα and ERβ) in the pathophysiology of breast cancer (BC) has considerably increased in last decades. Despite sharing a similar structure, these two transcription factors often exert opposite roles in BC. In addition, it has been shown that their transcriptional activity is not strictly associated to ligand activation and that unliganded ERs are able to "have a life on their own." This appears to be mainly due to ligand-independent mechanisms leading to ERs PTMs or to their recruitment to specific protein complexes, dependent on cellular context. Furthermore, a significant unliganded ER activity, probably independent by the activation of other pathways, has been recently reported to affect gene transcription, microRNA expression, and downstream proteome. In this review, we describe recent findings on nuclear and cytoplasmic unliganded ERα and ERβ activity. We focus on functional genomics, epigenomics, and interaction proteomics data, including PTM induced by ERs-modulated miRNAs in the BC context. A better comprehension of the molecular events controlled by unliganded ERs activity in BC pathogenesis is crucial since it may impact the therapeutic approach to the initial or acquired resistance to endocrine therapies, frequently experienced in the treatment of BC., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2016
- Full Text
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35. Thyroid development in zebrafish lacking Taz.
- Author
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Pappalardo A, Porreca I, Caputi L, De Felice E, Schulte-Merker S, Zannini M, and Sordino P
- Subjects
- Animals, Body Patterning genetics, Evolution, Molecular, Gene Expression Regulation, Developmental, Gene Knockdown Techniques, Larva growth & development, Larva metabolism, Morpholinos administration & dosage, Morpholinos genetics, Oligonucleotides, Antisense administration & dosage, Oligonucleotides, Antisense genetics, Organ Size genetics, Phylogeny, RNA, Messenger genetics, RNA, Messenger metabolism, Thyroid Gland growth & development, Thyroid Gland metabolism, Transcriptional Coactivator with PDZ-Binding Motif Proteins, Zebrafish growth & development, Intracellular Signaling Peptides and Proteins deficiency, Intracellular Signaling Peptides and Proteins genetics, Thyroid Gland embryology, Zebrafish embryology, Zebrafish genetics, Zebrafish Proteins deficiency, Zebrafish Proteins genetics
- Abstract
Taz is a signal-responsive transcriptional coregulator implicated in several biological functions, from chondrogenesis to regulation of organ size. Less well studied, however, is its role in thyroid formation. Here, we explored the in vivo effects on thyroid development of morpholino (MO)-mediated knockdown of wwtr1, the gene encoding zebrafish Taz. The wwtr1 gene is expressed in the thyroid primordium and pharyngeal tissue of developing zebrafish. Compared to mammalian cells, in which Taz promotes expression of thyroid transcription factors and thyroid differentiation genes, wwtr1 MO injection in zebrafish had little or no effect on the expression of thyroid transcription factors, and differentially altered the expression of thyroid differentiation genes. Analysis of wwtr1 morphants at later stages of development revealed that the number and the lumen of thyroid follicles, and the number of thyroid follicle cells, were significantly smaller. In addition, Taz-depleted larvae displayed patterning defects in ventral cranial vessels that correlate with lateral displacement of thyroid follicles. These findings indicate that the zebrafish Taz protein is needed for the normal differentiation of the thyroid and are the first to suggest that Taz confers growth advantage to the endocrine gland., (Copyright © 2015 Elsevier Ltd. All rights reserved.)
- Published
- 2015
- Full Text
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36. Cross-species toxicogenomic analyses and phenotypic anchoring in response to groundwater low-level pollution.
- Author
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Porreca I, D'Angelo F, Gentilcore D, Carchia E, Amoresano A, Affuso A, Ceccarelli M, De Luca P, Esposito L, Guadagno FM, Mallardo M, Nardone A, Maccarone S, Pane F, Scarfò M, Sordino P, De Felice M, and Ambrosino C
- Subjects
- Animals, Biomarkers, Environmental Exposure adverse effects, Female, Gene Expression, Gene Expression Profiling, Groundwater analysis, Liver drug effects, Liver metabolism, Liver Function Tests, Male, Mice, Reproducibility of Results, Species Specificity, Time Factors, Toxicity Tests, Acute, Toxicity Tests, Chronic, Zebrafish, Groundwater chemistry, Phenotype, Toxicogenetics, Water Pollution adverse effects
- Abstract
Background: Comparison of toxicogenomic data facilitates the identification of deregulated gene patterns and maximizes health risk prediction in human., Results: Here, we performed phenotypic anchoring on the effects of acute exposure to low-grade polluted groundwater using mouse and zebrafish. Also, we evaluated two windows of chronic exposure in mouse, starting in utero and at the end of lactation. Bioinformatic analysis of livers microarray data showed that the number of deregulated biofunctions and pathways is higher after acute exposure, compared to the chronic one. It also revealed specific profiles of altered gene expression in all treatments, pointing to stress response/mitochondrial pathways as major players of environmental toxicity. Of note, dysfunction of steroid hormones was also predicted by bioinformatic analysis and verified in both models by traditional approaches, serum estrogens measurement and vitellogenin mRNA determination in mice and zebrafish, respectively., Conclusions: In our report, phenotypic anchoring in two vertebrate model organisms highlights the toxicity of low-grade pollution, with varying susceptibility based on exposure window. The overlay of zebrafish and mice deregulated pathways, more than single genes, is useful in risk identification from chemicals implicated in the observed effects.
- Published
- 2014
- Full Text
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37. Bisphenol A interferes with thyroid specific gene expression.
- Author
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Gentilcore D, Porreca I, Rizzo F, Ganbaatar E, Carchia E, Mallardo M, De Felice M, and Ambrosino C
- Subjects
- Animals, Benzhydryl Compounds administration & dosage, Dose-Response Relationship, Drug, Endocrine Disruptors administration & dosage, Environmental Pollutants administration & dosage, Environmental Pollutants toxicity, Phenols administration & dosage, Pituitary Gland metabolism, Rats, Thyroid Gland cytology, Time Factors, Zebrafish, Benzhydryl Compounds toxicity, Endocrine Disruptors toxicity, Gene Expression Regulation drug effects, Phenols toxicity, Thyroid Gland drug effects, Thyroid Hormones genetics
- Abstract
Bisphenol A (BPA) is an endocrine-disrupting chemical that leads to low-dose human exposure due to its ability to leach from chemically derived products, as polycarbonate plastics and epoxy resin. In addition to its known xeno-endocrine action, BPA exerts a wide range of metabolic effects. Despite the documented BPA exposure outcomes on synthesis of thyroid hormones, there are not any data available on its actions on the thyroid follicular cells, site of synthesis of the thyroid hormones. Recently, it has been shown that several environmental pollutants, as BPA, can exert a thyroid disrupting activity. In this study, we employed in vitro and in vivo (zebrafish) models to examine the effects of BPA in regulating the expression of genes involved in the thyroid hormone synthesis and of their transcriptional regulators at BPA doses as low as 10(-9)M, a dose that is environmentally pertinent and far below the one detected in infants plasma. In both systems we could detect an altered expression of the genes involved in thyroid hormones synthesis and of thyroid specific transcriptional factors in BPA dose and time dependent manner. Our results suggest that BPA exerts a direct effect on thyroid follicular cell. We show that these cells can "sense" very low amount of BPA. Thus they, potentially, represent an ideal in vitro system to develop assays to detect BPA and other pollutants with thyroid disrupting activity at level far below the ones considered to be environmental relevant. Moreover, this report may provide new insight into the mode of BPA-induced deregulation of physiological processes as well as on the extensively debated molecular pathways underlying its biological activities., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)
- Published
- 2013
- Full Text
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38. Zebrafish bcl2l is a survival factor in thyroid development.
- Author
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Porreca I, De Felice E, Fagman H, Di Lauro R, and Sordino P
- Subjects
- Animals, Cell Survival, Morphogenesis, Thyroid Gland physiology, Transcription Factors physiology, Genes, bcl-2, Thyroid Gland embryology, Zebrafish embryology, Zebrafish Proteins physiology, bcl-X Protein physiology
- Abstract
Regulated cell death, defined in morphological terms as apoptosis, is crucial for organ morphogenesis. While differentiation of the thyroid gland has been extensively studied, nothing is yet known about the survival mechanisms involved in the development of this endocrine gland. Using the zebrafish model system, we aim to understand whether genes belonging to the Bcl-2 family that control apoptosis are implicated in regulation of cell survival during thyroid development. Evidence of strong Bcl-2 gene expression in mouse thyroid precursors prompted us to investigate the functions played by its zebrafish homologs during thyroid development. We show that the bcl2-like (bcl2l) gene is expressed in the zebrafish thyroid primordium. Morpholino-mediated knockdown and mutant analyses revealed that bcl2l is crucial for thyroid cell survival and that this function is tightly modulated by the transcription factors pax2a, nk2.1a and hhex. Also, the bcl2l gene appears to control a caspase-3-dependent apoptotic mechanism during thyroid development. Thyroid precursor cells require an actively maintained survival mechanism to properly proceed through development. The bcl2l gene operates in the inhibition of cell death under direct regulation of a thyroid specific set of transcription factors. This is the first demonstration of an active mechanism to ensure survival of the thyroid primordium during morphogenesis., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
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39. Oncogenic ras blocks the cAMP pathway and dedifferentiates thyroid cells via an impairment of pax8 transcriptional activity.
- Author
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Baratta MG, Porreca I, and Di Lauro R
- Subjects
- Animals, Cell Line, Cyclic AMP-Dependent Protein Kinases metabolism, Down-Regulation, Enhancer Elements, Genetic genetics, Humans, Kinetics, Mice, Organ Specificity genetics, PAX8 Transcription Factor, Protein Transport, Rats, Receptors, Thyrotropin metabolism, Symporters metabolism, Cell Dedifferentiation, Cyclic AMP metabolism, Genes, ras, Paired Box Transcription Factors metabolism, Signal Transduction, Thyroid Gland cytology, Transcription, Genetic
- Abstract
A deranged differentiation is often a landmark of transformed cells. We used a thyroid cell line expressing an inducible Ras oncoprotein in order to study the hierarchy of molecular events leading to suppression of thyroid-specific gene expression. We find that, upon Ras activation, there is an immediate global down-regulation of thyroid differentiation, which is associated with an inhibition of the cAMP signaling pathway. We demonstrate that an unusual negative cross talk between Ras oncogene and the cAMP pathway induces inactivation of the transcription factor Pax8 that we propose as a crucial event in Ras-induced dedifferentiation.
- Published
- 2009
- Full Text
- View/download PDF
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