Delta-Globin Gene Expression Is Enhanced in vivo by Interferon Type I

Beta hemoglobinopathies are widely spread monogenic lethal diseases. Delta-globin gene activation has been proposed as a possible approach for curing these pathologies. The therapeutic potential of delta-globin, the non-alpha component of Hemoglobin A2 (α2δ2; HbA2), has been demonstrated in a mouse model of beta thalassemia, while its anti-sickling effect, comparable to that of gamma globin, was established some time ago. Here we show that the delta-globin mRNA level is considerably increased in a Deoxyribonuclease II-alpha knockout mouse model in which type 1 interferon (interferon beta, IFNb) is activated. IFNb activation in the fetal liver improves the delta-globin mRNA level, while the beta-globin mRNA level is significantly reduced. In addition, we show that HbA2 is significantly increased in patients with multiple sclerosis under type 1 interferon treatment. Our results represent a proof of principle that delta-globin expression can be enhanced through the use of molecules. This observation is potentially interesting in view of a pharmacological approach able to increase the HbA2 level

Reversible Disruption of Pre-Pulse Inhibition in Hypomorphic-Inducible and Reversible CB1-/- Mice

Although several genes are implicated in the pathogenesis of schizophrenia, in animal models for such a severe mental illness only some aspects of the pathology can be represented (endophenotypes). Genetically modified mice are currently being used to obtain or characterize such endophenotypes. Since its cloning and characterization CB1 receptor has increasingly become of significant physiological, pharmacological and clinical interest. Recently, its involvement in schizophrenia has been reported. Among the different approaches employed, gene targeting permits to study the multiple roles of the endocannabinoid system using knockout (-/-) mice represent a powerful model but with some limitations due to compensation. To overcome such a limitation, we have generated an inducible and reversible tet-off dependent tissue-specific CB1-/- mice where the CB1R is re-expressed exclusively in the forebrain at a hypomorphic level due to a mutation (IRh-CB1-/-) only in absence of doxycycline (Dox). In such mice, under Dox+ or vehicle, as well as in wild-type (WT) and CB1-/-, two endophenotypes motor activity (increased in animal models of schizophrenia) and pre-pulse inhibition (PPI) of startle reflex (disrupted in schizophrenia) were analyzed. Both CB1-/- and IRh-CB1-/- showed increased motor activity when compared to WT animals. The PPI response, unaltered in WT and CB1-/- animals, was on the contrary highly and significantly disrupted only in Dox+ IRh-CB1-/- mice. Such a response was easily reverted after either withdrawal from Dox or haloperidol treatment. This is the first Inducible and Reversible CB1-/- mice model to be described in the literature. It is noteworthy that the PPI disruption is not present either in classical full CB1-/- mice or following acute administration of rimonabant. Such a hypomorphic model may provide a new tool for additional in vivo and in vitro studies of the physiological and pathological roles of cannabinoid system in schizophrenia and in other psychiatric disorders

In vivo activation of the human δ-globin gene: the therapeutic potential in β- thalassemic mice

β-thalassemia and Sickle Cell Disease are widespread fatal genetic diseases. None of the existing clinical treatments are resolving for all patients. So far two main strategies for the treatment are being investigated: (i) gene transfer of a normal β-globin gene; (ii) reactivation of the endogenous γ-globin gene. To date, neither approach has led to a satisfactory, commonly accepted standard of care. The δ-globin gene produces the δ-globin of the hemoglobin A2. Although low expressed, hemoglobin A2 is fully functional and could be a valid substitute of hemoglobin A in β-thalassemia disorder, as well as an antisickling agent in Sickle Cell Disease. Previous in vitro results suggested the feasibility to transcriptionally activate the human δ-globin gene promoter by inserting a Kruppel-like factor 1 binding site. We evaluate the activation of the Kruppel-like factor 1 containing δ-globin gene in vivo in transgenic mice. To evaluate the therapeutic potential we crossed the transgenic mice carrying a single copy activated δ-globin gene with a mouse model of β-thalassemia intermedia. Here we show that the human δ-globin gene can be activated in vivo in a stage and tissue specific fashion simply by the insertion of a Kruppel-like factor 1 binding site into the promoter. In addiction the activated δ-globin gene gives rise to a robust increase of the hemoglobin level in β-thalassemic mice, effectively improving the thalassemia phenotype. These results demonstrate, for the first time, the therapeutical potential of the δ-globin gene to treat severe hemoglobin disorders which could lead to novel approaches for the cure of β-hemoglobinopathies not involving gene addiction or reactivation

Differentiation of single cell derived human mesenchymal stem cells into cells with a neuronal phenotype: RNA and microRNA expression profile

The adult bone marrow contains a subset of non-haematopoietic cells referred to as bone marrow mesenchymal stem cells (BMSCs). Mesenchymal stem cells (MSCs) have attracted immense research interest in the field of regenerative medicine due to their ability to be cultured for successive passages and multi-lineage differentiation. The molecular mechanisms governing the self-renewal and differentiation of MSCs remain largely unknown. In a previous paper we demonstrated the ability to induce human clonal MSCs to differentiate into cells with a neuronal phenotype (DMSCs). In the present study we evaluated gene expression profiles by Sequential Analysis of Gene Expression (SAGE) and microRNA expression profiles before and after the neuronal differentiation process. Various tissue-specific genes were weakly expressed in MSCs, including those of non-mesodermal origin, suggesting multiple potential tissue-specific differentiation, as well as stemness markers. Expression of OCT4, KLF4 and c-Myc cell reprogramming factors, which are modulated during the differentiation process, was also observed. Many peculiar nervous tissue genes were expressed at a high level in DMSCs, along with genes related to apoptosis. MicroRNA profiling and correlation with mRNA expression profiles allowed us to identify putative important genes and microRNAs involved in the differentiation of MSCs into neuronal-like cells. The profound difference in gene and microRNA expression patterns between MSCs and DMSCs indicates a real functional change during differentiation from MSCs to DMSCs

Klf1 Affects DNase II-Alpha Expression in the Central Macrophage of a Fetal Liver Erythroblastic Island: a Non-Cell-Autonomous Role in Definitive Erythropoiesis▿

A key regulatory gene in definitive erythropoiesis is the erythroid Kruppel-like factor (Eklf or Klf1). Klf1 knockout (KO) mice die in utero due to severe anemia, while residual circulating red blood cells retain their nuclei. Dnase2a is another critical gene in definitive erythropoiesis. Dnase2a KO mice are also affected by severe anemia and die in utero. DNase II-alpha is expressed in the central macrophage of erythroblastic islands (CMEIs) of murine fetal liver. Its main role is to digest the DNA of the extruded nuclei of red blood cells during maturation. Circulating erythrocytes retain their nuclei in Dnase2a KO mice. Here, we show that Klf1 is expressed in CMEIs and that it binds and activates the promoter of Dnase2a. We further show that Dnase2a is severely downregulated in the Klf1 KO fetal liver. We propose that this downregulation of Dnase2a in the CMEI contributes to the Klf1 KO phenotype by a non-cell-autonomous mechanism

Picta fragmenta. La pittura vesuviana: una rilettura

Il pi\uf9 grande archivio della pittura antica: Pompei e la regione vesuviana lo sono ancora, a quasi tre secoli dall\u2019inizio della loro riscoperta. Anche se in questa seconda vita non tutto quello che era stato riportato alla luce ha superato la prova del tempo: molte decorazioni, ancora ben leggibili e ricche di colore al momento del rinvenimento, oggi sono scomparse. Per assicurare anche a chi verr\ue0 dopo di noi la conoscenza di questo eccezionale patrimonio, tanto prezioso quanto fragile, due sono le vie principali da percorrere: la conservazione della materia e il restauro della memoria. Solo attraverso azioni sistematiche di manutenzione i picta fragmenta di Pompei e gli altri vesuviani, cos\uec come quelli di tutti gli altri siti archeologici che hanno restituito tracce delle pareti dipinte del mondo antico, possono resistere all\u2019usura del tempo. Altrettanto importante \ue8 il lavoro di chi cerca di ricondurre i frammenti alla loro situazione originaria, ricomponendo disegni e restituendo colori, con un restauro che pu\uf2 essere reale e materico, ma anche virtuale. Alla base di entrambi sta l\u2019indispensabile miniera di informazioni che generazioni di ricercatori (archeologi, storici dell\u2019arte, archeometri) e di artisti (fotografi, disegnatori, pittori) hanno contribuito a formare. Da questa sinergia di competenze e di indirizzi nasce quell\u2019approccio globale e olistico allo studio della pittura antica che la comunit\ue0 scientifica indica come necessario: per meglio conoscere le pareti antiche in s\ue9, quale espressione di un\u2019arte applicata, e nei loro contesti, e per meglio farle conoscere. Pompei e i siti vesuviani sono, anche in questo, uno straordinario osservatorio-laboratorio: rileggere, con occhi nuovi e nuove domande, quello che delle loro pareti dipinte \ue8 arrivato sino a noi \ue8 l\u2019intento di questo libro. Alla ricerca, a partire dai picta fragmenta, delle picturae antiche: una realt\ue0 al plurale, da indagare con un approccio al plurale