1,721,668 research outputs found
Foetal haemoglobin inducers and thalassaemia: novel achievements
No full textBeta-thalassaemias are a group of hereditary human diseases caused by more that 200 mutations of the human β-globin gene, leading to low or absent production of adult β-globin and an excess of α-globin, causing ineffective erythropoiesis and low or absent production of adult haemoglobin (HbA)1. The conventional treatment for these patients is based on regular blood transfusions and chelating therapy
New trends in the development of transcription factor decoy (TFD) pharmacotherapy
No full textMany recent published observations firmly demonstrate that one of the ways to study and artificially modulate gene expression at the transcriptional level is offered by the "transcription factor decoy" (TFD) strategy. This experimental approach is based on the competition for trans-acting factors between endogenous cis-elements present within regulatory regions of target genes and exogenously added DNA sequences (the DNA-based drug) mimicking the specific cis-elements. The objective of this molecular intervention is to cause a decrease of the interactions of trans-factors with the target genomic cis-elements, leading to alteration of transcription. The characterisation of the biological activity of the designed decoy molecules is routinely assessed by molecular technologies, such as electrophoretic mobility gel shift assay (EMSA), competitive DNase I footprinting, in vitro transcription. New advances in this field employ biospecific interaction analysis (BIA) based on surface plasmon resonance (SPR) and biosensor technology. With respect to the design of the decoy biomolecules, in addition to double-stranded DNA/DNA hybrids, cross-linking between two DNA molecules either via photocrosslinking or by the introduction of a covalently linked, non-nucleotide bridge has been reported. Furthermore, RNA decoys have been described able to bind transcription factors via aptameric interactions. In addition, circular decoys assuming a dumbbell configuration or single-stranded decoys with intramolecular palindromic sequences have also been described. Decoy molecules were also produced by polymerase-chain reaction (PCR). More recently, peptide nucleic acids-DNA chimeras have been shown to exhibit decoy activity and high level of stability. This variety of decoy biomolecules facilitate the establishment of suitable delivery approaches, including pressure-mediated transfer, electrically enhanced transfer, biolistic bombardment, cationic liposomes, hemagglutinating virus of Japan (HVJ)-liposomes, microsphere-aided delivery, nano-particles, peptide-mediated delivery, steroid mediated gene transfer, and red-blood cells
Hemoglobin production in beta-thalassemia erythroid cells following alteration of biomolecular pathways regulating globin gene expression. Durata: 24 mesi (dal 26/11/2010 al 26/11/2012). Progetto n°: GGP10214. Finanziato da: TELETHON 2010.
No full textThe general aim of this proposal is to analyse the gene expression of erythroid progenitor cells from
beta−thalassemia patients, for the identification of approaches enhabling the induction of fetal
hemoglobin (HbF) and production of adult hemoglobin (HbA), of possible interest for the therapy of
beta−thalassemia. The objective of Task 1 is the recruitment and genetic characterization of the
beta−thalassemia patients involved in this study. The objective of Task 2 is the characterization of the
molecular mechanisms of action of novel HbF inducers and their use in combination with gene therapy
employing lentiviral vectors. The objective of Task 3 is the study of gene expression in HPFH patients
and in beta−thalassemia patients producing very high levels of HbF. The objective of Task 4 is the
analysis of micro RNA network in HPFH cells and in erythroid progenitors induced to HbF
production. The objective of Task 5 is the application of the read−through approach on HbA
production by beta°39 thalassemic cells and transgenic animals. The objective of Task 6 is the study of
the effects of siRNAs or antisense DNA or DNA−PNA chimeras on target mRNA sequences. Task 7 is
dedicated to exploitation, for the identification of Biotech companies interested in the developed
protocols and patents
MicroRNA and erythroid differentiation
No full textMicroRNA (miRNAs, miRs) are a family of small noncoding RNAs that regulate gene expression by sequence-selective targeting if miRNAs, leading to translational repression or mRNA degradation. Considering that a single miRNA can target several mRNAs and the 3'UTR sequence of a single mRNA might contain several signals for miRNA recognition, at least 10-40% of human mRNAs are potential targets of microRNAs, leading to control of highly regulated biological functions. In our laboratory we have analyzed by microarray the miR-profile in erythroid precursor cells (ErPC) from normal and thalassemic patients expressing different levels of foetal haemoglobin (including patients exhibiting a HPFH phenotype). The microarray data were confirrmed by RT-PCR analysis, and allowed us to identify miR-210 as highly expressed in the erythroid precursor cells from HPFH patients. When RT-PCR was performed on mithramycin (MTH)-induced K562 cells and erythroid precursor cells, we demonstrated that miR-210is induced in time-dependent and dose-dependent fashion, together with induction of gamma-globin genes. Molecular biology studies allowed to identify raptor mRNA and TORC1 as putative miR-210 targets. As far as modulation of miR-210, peptide nucleic acids (PNAs) might be of interest. We have studies a PNA conjugated to a polyarginine peptide which (a) is efficiently internalized within target cells; (b) strongly inhibits miR-210 activity; (c) deeply alters the expression of raptor mRNA and gamma-globin gene
Targeting the MicroRNAs-Transcription Factors Network for Innovative Therapeutic Protocols for ???-Thalassemia and Sickle-cell Anemia
No full textMicroRNAs (miRNAs, miRs) are a family of small non-coding RNAs
that regulate gene expression by targeting mRNAs in a sequence
specific manner, inducing translational repression or mRNA
degradation. MicroRNAs have been found deeply involved in the
control of erythroid differentiation. In human trisomy 13, there is
delayed switching and persistence of HbF and elevation of embryonic
hemoglobin in newborns (Sankaran et al., 2001). By examining the
genes in this region, two microRNAs, miR-15a and miR-16-1, appear
as top candidates for the elevated HbF levels. Increased expression of
these microRNAs in primary human erythroid progenitor cells results in
elevated fetal and embryonic hemoglobin gene expression. The target of
these microRNAs was identified in MYB mRNA; interestingly, MYB
plays an important role in silencing the fetal and embryonic hemoglobin
genes. The microRNA miR-486-3p regulates BCL11A expression by
binding to the extra-long isoform of BCL11A 3'UTR (Lulli et al., 2013). Overexpression of miR-486-3p in erythroid cells resulted in reduced BCL11A protein levels, associated to increased expression of beta-globin gene, whereas inhibition of physiological miR-486-3p levels increased BCL11A and, consequently, reduced beta-globin expression. The data obtained indicate that BCL11A, one of the major repressor of
beta-globin gene expression, is a molecular target of miR-486-3p;
accordingly, pharmacological mediated up-regulation of miR-486-3p
might lead to BCL11A down-regulation and, consequently activation of
the beta-globin gene expression. Other microRNAs found up-regulated in
association with beta-globin gene expression were miR-210 (Bianchi et al., 2009), miR-26b (Alijani et al., 2014), miR-451 (Kouhkan et al., 2014). As a first conclusion, the findings that microRNAs are involved in beta-globin anticipate the possibility that their pharmacological alteration
might be a key strategy for increase HbF in erythroid cells
Predictive Analyses of Biological Effects of Natural Products: From Plant Extracts to Biomolecular Laboratory and Computer Modeling
No full textYear by year, the characterization of the biological activity of natural products is becoming more competitive and complex, with the involvement in this research area of experts belonging to different scientific fields, including chemistry, biochemistry, molecular biology, immunology and bioinformatics. These fields are becoming of great interest for several high-impact scientific journals, including eCAM. The available literature in general, and a survey of reviews and original articles recently published, establishes that natural products, including extracts from medicinal plants and essential oils, retain interesting therapeutic activities, including antitumor (1-7), antiviral (8), anti-inflammatory (9), pro-apoptotic (10,11) and differentiating (12) properties. In this commentary, we focus attention on interest in networks based on complementary activation and comparative evaluation of different experimental strategies applied to the discovery and characterization of bioactive natural products. A representative flow chart is shown in Fig. 1
Alternative options for DNA-based experimental therapy of β-thalassemia.
No full textINTRODUCTION: Beta-thalassemias are caused by more than 200 mutations of the β-globin gene, leading to low or absent production of adult hemoglobin. Achievements have been made with innovative therapeutic strategies for β-thalassemias, based on research conducted at the levels of gene structure, transcription, mRNA processing and protein synthesis. AREAS COVERED: The objective of this review is to describe the development of therapeutic strategies employing viral and non-viral DNA-based approaches for treatment of β-thalassemia. EXPERT OPINION: Modification of β-globin gene expression in β-thalassemia cells has been achieved by gene therapy, correction of the mutated β-globin gene and RNA repair. In addition, cellular therapy has been proposed for β-thalassemia, including reprogramming of somatic cells to generate induced pluripotent stem cells to be genetically corrected. Based on the concept that increased production of fetal hemoglobin (HbF) is beneficial in β-thalassemia, DNA-based approaches to increase HbF production have been optimized, including treatment of target cells with lentiviral vectors carrying γ-globin genes. Finally, DNA-based targeting of α-globin gene expression has been applied to reduce the excess of α-globin production by β-thalassemia cells, one of the major causes of the clinical phenotype
Thalassaemia modular stratification system for personalized therapy of beta-thalassemia (THALAMOSS). Durata: 48 mesi (dal 01/11/2011 al 31/12/2015). Progetto n°: 306201. Finanziato da: FP7-HEALTH-2012-INNOVATION-1
No full textTHALAMOSS is aimed at development of universal sets of markers and techniques for stratification of
β-thalassaemia patients into treatment subgroups for (a) onset and frequency of blood transfusions, (b) choice
of iron chelation, (c) induction of fetal hemoglobin, (d) prospective efficacy of gene-therapy. At present, no
framework exists to guide therapeutic decisions and personalised treatment of β-thalassaemia.
THALAMOSS Workpackages: WP1. Recruitment, patient characterization and development of erythroid
precursor cells cultures; WP2. Omics analyses; WP3. Novel therapeutic approaches; WP4. Data analysis; WP5.
Dissemination and exploitation; WP6. Regulatory and ethical issues; WP7. Management.
The impact of THALAMOSS is the provision of novel biomarkers for distinct treatment subgroups in
β-thalassaemia (500-1000 samples from four European medical centres), identified by combined genomics,
proteomics, transcriptomics and tissue culture assays, and establishment of routine techniques for detection of
these markers. Translation of these activities into the product portfolio and R&D methodology of participating
SMEs will be a major issue. THALAMOSS tools and technologies will (a) facilitate identification of novel
diagnostic tests, drugs and treatments specific to patient subgroups and (b) guide conventional and novel
therapeutical approaches for β-thalassaemia, including personalised medical treatments
Titolo del progetto: MicroRNA Therapeutics in CF: Targeting CFTR and inflammation networks (MicroRNA-CF)
No full textEpigenetic regulation of pro-inflammatory genes and direct regulation of CFTR protein expression by MicroRNA has been reported. The objectives of the project MicroRNA-CF are: identification of specific MicroRNAs and their most relevant mRNA targets in CF bronchial epithelial cells; study of changes of the gene expression and secretome profile of CF using PNAs targeting MicroRNAs; development and biological validation of novelPNA-based molecules interfering with MicroRNAs involved in inflammation; development of PNA-based antagomiRNAs for CFTR stabilization
Design and synthesis of improved analogs of trimethylangelicin (TMA) for personalized treatment of cystic fibrosis. Durata: 12 mesi (dal 01/09/2014 al 31/08/2015) Progetto n°: FFC#8/2014. Finanziato da: Fondazione per la ricerca sulla Fibrosi Cistica-Bando 2014
No full textThe major objective of the project is to test TMA analogs with the aim offinding new antinflammatory agents and/or CFTR function modulators with equal or higher activity in respect to the parent TMA, and with absent DNA photobinding properties. In addition, the newly synthesized compounds will allow to derive structure-activity relationships on both the biological activities on inflammation and CFTR modulation. The goals of the project can be outlined in 5 sections: 1) design and synthesis of new TMA analogs; 2) test of photoreactivity; 3) test of the anti-inflammatory activity, based on inhibition of NFkB/DNA interaction and expression of pro-inflammatory genes in CF bronchial cell lines and 4) test of the effects as CFTR function modulation; 5) derivation of structure-activity relationships aimed at rationalizing the structural determinants required to obtain selective anti-inflammatory properties, selective CFTR modulatory properties and dual antiinflammatory/ CFTR modulatory activity
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