Project description:This study uses microarray technology to examine the erythroid progenitor mRNA of patients with transfusion dependent β-thalassaemia and compare it to erythroid progenitor mRNA from healthy controls. We observed no statistical difference in gene expression between the groups following 7 days in culture. However, following 14 days in culture we observed differential expression of 161 genes. Haematopoietic cells from the peripheral blood of 6 β-thalassaemia patients and 6 healthy controls was grown in semi-solid media. After 7 and 14 days in culture cells of erythroid origin were isolated. Total RNA was isolated from these for microarray gene expression analysis
Project description:This study uses microarray technology to examine the erythroid progenitor mRNA of patients with transfusion dependent ?-thalassaemia and compare it to erythroid progenitor mRNA from healthy controls. We observed no statistical difference in gene expression between the groups following 7 days in culture. However, following 14 days in culture we observed differential expression of 277 genes. Haematopoietic cells from the peripheral blood of 6 ?-thalassaemia patients and 6 healthy controls was grown in semi-solid media. After 7 and 14 days in culture cells of erythroid origin were isolated. Total RNA was isolated from these for microarray gene expression analysis
Project description:This study uses microarray technology to examine the erythroid progenitor mRNA of patients with transfusion dependent β-thalassaemia and compare it to erythroid progenitor mRNA from healthy controls. We observed no statistical difference in gene expression between the groups following 7 days in culture. However, following 14 days in culture we observed differential expression of 161 genes.
Project description:This study uses microarray technology to examine the erythroid progenitor mRNA of patients with transfusion dependent β-thalassaemia and compare it to erythroid progenitor mRNA from healthy controls. We observed no statistical difference in gene expression between the groups following 7 days in culture. However, following 14 days in culture we observed differential expression of 277 genes.
Project description:Reactivation of gamma-globin is considered a promising approach for the treatment of beta-thalassaemia and sickle cell disease. Therapeutic induction of gamma-globin expression is fraught with lack of suitable therapeutic targets. In order to identify new potential targets we analysed the changes in the proteome of human primary erythroid progenitor cells by treatment with decitabine, a known, yet not clinically safe, gamma-globin inducer. Significant differentially expressed proteins were identified which were involved in various biological pathways and functional categories.
Project description:The Affymetrix Human Gene 2.0 ST array was used to measure differential expression of RNA isolated from normal and Diamond Blackfan anemia (DBA) erythroid progenitors after ex vivo expansion of circulating, peripheral blood derived hematopoietic stem cells under erythroid growth conditions. The gene-level probe summaries reported in this series were computed using RMA as implemented in the Bioconductor package Oligo v1.36.1. Diamond Blackfan anemia (DBA) is a congenital bone marrow failure syndrome characterized by erythroid aplasia, usually without perturbation of other hematopoietic lineages. Approximately 65% of DBA patients with autosomal dominant inheritance have heterozygous mutations or deletions in ribosomal protein (RP) genes while <1% of patients with X-linked inheritance have been identified with mutations in the transcription factor, GATA1. Erythroid cells from patients with DBA have not been well characterized and the mechanisms underlying the erythroid specific effects of either RP or GATA1 associated DBA remain unclear. We have developed an in vitro culture system to expand peripheral blood CD34+ progenitor cells from patients with DBA and differentiate them into erythroid cells. Cells from patients with RP or GATA1 mutations showed decreased proliferation and delayed erythroid differentiation compared to controls. RNA transcript analyses of erythroid cells from controls and patients with RP or GATA1 mutations showed distinctive differences, with upregulation of heme biosynthesis genes prominently in RP-mediated DBA and failure to upregulate components of the translational apparatus in GATA1-mediated DBA. Our data show that dysregulation of translational function is a common feature of DBA caused by both RP and GATA1 mutations.
Project description:RNA-seq was performed using RNA extracted from enriched T-cell blasts for the following study groups: healthy controls (NC, n=5) and patients with AIOLOS Q402* (n=3 ). Illumina libraries were generated using the SMARTer Stranded Total RNA-Seq Kit v3 – Pico Input Mammalian Components (Takara San Jose, CA). Libraries were sequenced on a Illumina HiSeq 2500 instrument using 2 x 75 bp paired-end reads, and alignment and mapping analyses were performed by the Takara Cogent NGS Analysis Pipeline v1.5.0 (also known as CogentAP). Differential gene expression analysis was performed by DESeq2 v1.32.0 [Bioconductor version: Release (3.15)].
Project description:RNA-seq was performed using RNA extracted from EBV-immortalized B-cells for the following study groups: healthy controls (HC, n=3) and patients with AIOLOSE 82K (n=4 ). Illumina libraries were generated using the SMARTer Stranded Total RNA-Seq Kit v3 – Pico Input Mammalian Components (Takara San Jose, CA). Libraries were sequenced on a Illumina NextSeq 2000 instrument using 2 x 75 bp paired-end reads, and alignment and mapping analyses were performed by the Takara Cogent NGS Analysis Pipeline v1.5.0 (also known as CogentAP). Differential gene expression analysis was performed by DESeq2 v1.32.0 [Bioconductor version: Release (3.15)].
