Project description:Lifelong treatment and major complications characterize β-thalassemia patients, whether they suffer from the milder type of the disease (β-thalassemia intermedia, TI) or the severe type (β-thalassemia major, TM). This study aimed to ascertain the differences in the transcriptional profiles of TI and TM patients to identify targets for stratification and the development of personalized therapeutic strategies. RNA-seq of 49 participants (16 TI, 16 TM, 17 healthy) identified a higher number of differentially expressed genes in TM than in TI patients when compared to healthy participants portraying accurately the clinical observations. When TI patients were compared to TM, the absence of significantly differentially expressed genes highlighted the increased variability seen in the TI expression profiles suggesting that a continuous spectrum describes the disease and not distinct conditions. Although no confounding effect was seen due to different centers, age groups and various clinical characteristics, when male and female patients were compared to healthy participants of the same gender, it was evident that males and females are affected in distinctive ways by β-thalassemia. TI triggers a pronounced and consistent transcriptional signature in males, whereas a small effect is seen in females reflecting increased biological variability. TM induces transcriptional changes of similar magnitude in both genders with limited overlap between them emphasizing that different pathways are affected. Our study provides a framework for further stratification studies in β-thalassemia and highlights the need to consider gender as an important variable for developing differential diagnostic and therapeutic strategies.
Project description:To explore the role of circRNAs in the regulation of β-thalassemia and provide new evidence and targets for the treatment of β-thalassemia, circRNAs expression profiling was analyzed from patients with β-thalassemia intermedia and major.
Project description:a comparison of a control and a mutant condition (Th3) of the Hemoglobin Beta major and minor chain, used as Beta Thalassemia mouse model. 4 controls and 4 mutants
Project description:To explore the relationship between miRNAs expression and pediatric patients with β-thalassemia, we analyzed abnormal expressed miRNAs in peripheral blood of pediatric β-thalassemia by miRNA sequencing.
Project description:a comparison of a control and a mutant condition (Th3) of the Hemoglobin Beta major and minor chain, used as Beta Thalassemia mouse model.
Project description:β-thalassemia cell lines were generated via CRISPR-Cas9 genome editing of Bristol Erythroid Line Adult (BEL-A) and differentiated to the basophilic and polychromatic erythroid cell stage. TMT comparative proteomics was then performed on stage matched WT and β-thalassemia cells isolated by FACS.
Project description:β-thalassemia major can be caused by homozygous mutations of the HBB gene, most of the cases are inherited from parents who both have β-thalassemia minor. Herein, we show that a mosaic paternal uniparental isodisomy of chromosome 11p14.3-15.5 is associated with β-thalassemia major in a patient with β-thalassemia minor-that evolved to β-thalassemia major. From this case, we suggest that analysis of HBB gene for non-hematopoietic tissues should be performed in late-onset β-thalassemia major patients. Keywords: genomic
Project description:Background: The thalassemias are highly diverse at both the molecular and clinical levels. Many of the HBB mutations that result in β-thalassemia are missense mutations in the coding region of the β-globin gene, but a few cause alternative splicing, and interfere with normal processing of the β-globin transcripts. Transcriptome profiling in individuals affected with β-thalassemia, especially in individuals who carry novel mutations in the HBB, may improve our understanding of the heterogeneity and molecular mechanisms of the disease. Methods: Members of a family with a daughter affected with thalassemia intermedia, although her mother was not clinically affected, were examined for physical characteristics, hematological parameters and β-globin gene sequences. We also characterized genome-wide gene expression in the family using RT-qPCR and high-throughput RNA-sequencing mRNA expression profiling of blood. Results: Clinical findings, hematological indices, DNA and RNA sequence analysis of individuals with β-thalassemia, including the description of a novel mutation in the β-globin gene, which introduces a cryptic donor splice site. More than 300 genes are differentially expressed in β-thalassemic blood with many of the DEGs involved in pathways relevant to the clinical management of β-thalassemia. β-thalassemia shows important similarities and differences with sickle cell disease at the transcriptome level. Conclusions: We described the down-regulation of the β-globin gene in β-thalassemia by RNA-sequencing analysis using a sample from an affected individual and her mother, who have a novel mutation in the HBB that creates a cryptic donor splice site. The daughter has a typical β-thalassemia allele as well, and an unexpectedly severe phenotype. The DEGs are enriched in pathways that are directly or indirectly related to β-thalassemia such as hemopoiesis, heme biosynthesis, response to oxidative stress, inflammatory responses, immune responses, control of circadian rhythm, apoptosis, and other cellular activities. We compare our findings with published results of RNA-Sequencing analysis of sickle cell disease (SCD) and erythroblasts from a KLF1-null neonate with hydrops fetalis, and recognize similarities and differences in their transcriptional expression patterns.
Project description:β-thalassemia major can be caused by homozygous mutations of the HBB gene, most of the cases are inherited from parents who both have β-thalassemia minor. Herein, we show that a mosaic paternal uniparental isodisomy of chromosome 11p14.3-15.5 is associated with β-thalassemia major in a patient with β-thalassemia minor-that evolved to β-thalassemia major. From this case, we suggest that analysis of HBB gene for non-hematopoietic tissues should be performed in late-onset β-thalassemia major patients. Experiment Overall Design: This study is to evaluate the cause of delay-onset β-thalassemia major in our patient. Patients peripheral blood, hair follicle, and oral mucosa, and her parents pripheral blood samples were analyzed.
Project description:We used a modified 5i/L/FA system to generate transgene-free naïve iPSCs directly from the fibroblasts of a patient suffering from β-thalassemia and further demonstrated efficient gene correction with a CRISPR/Cas9 system, which provides an improved strategy for personalized treatment of β-thalassemia.