Il giorno 14 Aprile 2016, alle ore 15, si terrà, presso l’Aula 27 dell’ed. A dell’Area della Ricerca del CNR di Pisa, Via G. Moruzzi 1, un seminario dedicato alle attività scientifiche laboratorio Zebrafish, una delle facilities di IFC per lo studio di patologie umane su modelli animali. Il seminario si articola in 5 relazioni:
- Animal models of human disease: zebrafish swim into view (Letizia Pitto, IFC)
- Zebrafish as a model for studying congenital heart diseases (Elena Guzzolino, PhD Scuola S.Anna-IFC)
- Zebrafish as a tool for identifying new therapeutic targets for melanoma treatment (Samanta Sarti, PhD Università di Siena-ITT)
- MicroRNA mir-29 controls a compensatory response to limit neuronal iron accumulation during adult life and aging (Roberto Ripa, PhD Scuola Normale Superiore Pisa)
- Zebrafish: a tool for in vivo studies on human muscular dystrophy and epilepsy (Maria Marchese Molecular Medicine, IRCCS Stella Maris)
Brevemente si ricorda l’esperienza professionale della Dott.ssa Pitto. Ricercatore dell’Istituto di Fisiologia Clinica del CNR Pisa dal 1990, nel decennio 1992-2001 è responsabile del corso “Cellule somatiche e rigenerazione” e coordinatore dell’area di "Tecnologie Vegetali ed Ingegneria Genetica” per la Scuola di Specializzazione in Biotecnologie vegetali dell’Università di Pisa. Tra il 1996 e il 2001 è Membro del Comitato Scientifico dell’allora Istituto di Mutagenesi e Differenziamento del CNR, oggi integrato in IFC, ricoprendo anche il ruolo di Group leader del laboratorio di Differenziamento Vegetale dell’IMD.
Dal 2005 è Responsabile del corso "Ruolo dei microRNA nello sviluppo e nelle patologie" all’interno del corso di Ingegneria Genetica della Facoltà di Scienze Biologiche della Università di Pisa. Dal 2007 è Group leader dell’ "RNA interference Lab” dell’IFC, per il quale ricopre dal 2009 anche il ruolo di Responsabile della Zebrafish facility. Ha 60 reports ISI web of knowledge con H index =17, il n° di citazioni totale = 968 ed una media di citazioni per articolo di 13.08. Il suo IF totale è di 210,63 e quello medio di 3,57 come risulta da Research Gate.
Di seguito gli Abstract dei seminari.
Animal models of human disease: zebrafish swim into view
Letizia Pitto IFC
Biomedical research depends on the use of animal models to understand the pathogenesis of human diseases at a cellular and molecular level and to provide systems for developing and testing new therapies. Despite the pre-eminence of the mouse in modeling human diseases, several aspects of murine biology limit its routine use in large-scale genetic and therapeutic screening. Many researchers who are interested in an embryologically and genetically tractable disease model have now turned to zebrafish. Zebrafish biology allows ready access to all developmental stages, and the optical clarity of embryos and larvae allow real-time imaging of developing pathologies. The IFC zebrafish lab which is responsible for supporting different research groups working in cardiovascular diseases, neuromuscular disorders, aging and cancer is a good example of the plasticity of this animal model. Some examples of the studies which are in progress in the IFC zebrafish lab will be presented in the seminar.
Zebrafish as a model for studying congenital heart diseases
Elena Guzzolino PhD Scuola S.Anna-IFC
Tbx5 is a gene of the T-box transcription factor family that has a key role in heart/limb development of vertebrate. In humans mutations against Tbx5 are responsible of Holt-Oram syndrome (HOS) a congenital autosomal dominant pathology with an incidence of 1/100000 birth. Among the many genes regulated by Tbx5 our work has focused on miRNAs, negative regulators of gene expression at post-transcriptional level. We performed a miRNA-profiling on RNA extracted from E11.5–E12.0 hearts isolated from WT and HOS mice, identified differentially expressed miRNAs and functionally tested them in zebrafish. We focused on miR-182, which was recently identified as a potential prognostic marker in cardiac heart failure and is overexpressed in patients with coronary artery disease. Overexpression of miR-182 affects proliferation and migration of myocardial cells during the first stages of zebrafish development and induces arrythmies, a cardiac defect which often characterizes HOS patients.
Zebrafish as a tool for identifying new therapeutic targets for melanoma treatment.
Samanta Sarti PhD Università di Siena-ITT
Melanoma is the most aggressive form of skin cancer, with poor prognosis at advanced stage. BRAF is a protein kinase belonging to the highly oncogenic MAPK pathway and it is mutated in about 50% of melanoma cases. The most common mutation is the V to E aminoacidic substitution at the 600 residue, which leads to the formation of a constitutively active BRAFV600E protein. This mutation is also present in benign melanocytic naevi suggesting the presence of additional genetic alterations at the basis of melanoma formation. To identify and study functional interactors of BRAFV600E that could to contribute to melanoma development, and hence that could to be new therapeutic targets, we are using a specific zebrafish melanoma model. We are taking advantage from the Tg(mitfa:BRAFV600E);p53-/-;mitfa-/- transgenic zebrafish line and the miniCoopR system which allow to study the role of a gene of interest in melanoma progression and not only in melanomagenesis.
MicroRNA mir-29 controls a compensatory response to limit neuronal iron accumulation during adult life and aging.
Roberto Ripa PhD Scuola Normale Superiore Pisa
Intracellular iron homeostasis is strictly controlled by two RNA binding proteins known as Iron Regulatory Proteins (IRP1 and IRP2) that regulate at post-transcriptional level the expression of iron management genes optimizing cellular iron availability. Targeted deletions of IRPs proteins in animals have demonstrated that IRP2 plays a pivotal role in regulating iron homeostasis in the Central Nervous System. Despite this fine regulation, impairment of iron homeostasis occurs during aging and in many neurodegenerative diseases, as consequence, iron progressively accumulates and leads to increased oxidative stress and toxicity and the underlying mechanisms are still unclear. We investigated the age-dependent changes of iron homeostasis using zebrafish and the short lived fish Nothobranchius furzeri.
Our studies indicate that mir-29 modulates intracellular iron homeostasis and is up-regulated as adaptive response to limit excessive iron accumulation, to prevent early-onset aging processes.
Zebrafish: a tool for in vivo studies on human muscular dystrophy and epilepsy
Maria Marchese Molecular Medicine, IRCCS Stella Maris
Recently zebrafish became a valuable tool for neurobehavioral studies because larvae display several, quantifiable behavioral phenotypes that may recapitulate human behaviors. Zebrafish have been used to study muscular dystrophy including dystrophin deficiency and different forms of dystroglycanopathy. This small vertebrate have muscle structure and development similar to that of mammals, and the majority of muscle-related genes tested are present in zebrafish. We propose that additional dystrophic mutants will provide models for other dystrophies, and to uncover additional dystrophy-associated genes that will further our understanding of the pathology of muscular dystrophy. Zebrafish is also excellent to studying epilepsy. It was demonstrated that using behavioral, molecular, pharmacological and electrophysiological techniques, immature zebrafish have seizures reproducing many of the crucial aspects of mammalian seizures. Given the conserved structure and function of many zebrafish homologs of human epilepsy genes, we used a new approach of gene editing in zebrafish which could yield fundamental insights as well as new therapies.