MiRNA expression profile analysis in myelodysplastic syndromes
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ABSTRACT: The miRNA-related signaling pathways in MDS could be screened using high throughput bioinformatics analysis based on the miRNAs expression profile network. Here, we tried to identify the miRNAs-regulated pathways through a miRNA microarray in CD34+ cells from MDS patients. miRNA expression profile analysis was performed in 12 MDS patients and 6 normal controls using GeneChip® miRNA 3.0 Array, and diffirential miRNAs were identified.
Project description:The pathogenesis-related signaling pathways in MDS could be screened using high throughput bioinformatics analysis based on the gene expression profile network. Here, we tried to identify the mRNAs-regulated pathways through a mRNA microarray in CD34+ cells from MDS patients. Gene expression profile analysis was performed in 12 MDS patients and 6 normal controls using GeneChip® PrimeView� Human Gene Expression Array, and diffirential genes were identified.
Project description:The progressive mechanism of myelodysplastic syndrome (MDS) remains unknown. We report that ROBO1 and ROBO2 are identified as novel progression-related somatic mutations using whole-exome and targeted sequencing in six of 16 (37.5%) paired MDS patients undergoing disease progression. To investigated the effect of ROBO1 or ROBO2 on ROBO1/2 CN number and LOH, we employed a Cytosan 750K chip to analyze the copy-number variations (CNVs) and loss of heterogeneity (LOH) in MDS patients with ROBO1&2 mutations. Copy number and LOH analysis of Affymetrix CytoScan 750K array was performed for 14 MDS patients with ROBO1 or ROBO2 mutations
Project description:We investigated the spectra of circulating miRNAs in plasma of myelodysplastic syndromes (MDS) patients. Peripheral blood plasma from MDS patients with different risk scores was used for Agilent miRNA expression microarray analysis to define miRNA profile and to find miRNAs with discriminatory levels for lower risk and higher risk MDS. Results were further validated using droplet digital PCR on a larger cohort, enabling absolute quantification of plasma miRNAs and defining miRNAs with prognostic value for the disease. We analyzed expression profile of circulating miRNAs in plasma from 21 individuals: 7 controls and 14 MDS patients.
Project description:Control of oxidative stress in the bone marrow (BM) is key for maintaining the balance between self-renewal, proliferation, and differentiation of hematopoietic cells. Breakdown of this regulation can lead to diseases characterized by BM failure such as the myelodysplastic syndromes (MDS). To better understand the role of oxidative stress in MDS development, we compared protein carbonylation as an oxidative stress marker in BM of patients with MDS and control subjects, and also patients with MDS under treatment with the iron chelator deferasirox.
Project description:The miRNA-related signaling pathways in MDS could be screened using high throughput bioinformatics analysis based on the miRNAs expression profile network. Here, we tried to identify the miRNAs-regulated pathways through a miRNA microarray in CD34+ cells from MDS patients.
Project description:Myelodysplastic syndromes (MDS) are a heterogenous group of hematopoietic stem cell disorders characterized by dysplastic blood cell formation and peripheral blood cytopenias. Up to 30% of patients with MDS will progress to a highly chemotherapy-resistant secondary acute myeloid leukemia (sAML). We identified mutations in U2AF1 in MDS patients and patients with U2AF1 mutations are at an increased risk of developing sAML. We identified mutations in U2AF1 in patients with MDS and hypothesized that U2AF1 mutations may represent a novel mechanism that could alter gene expression in MDS. To elucidate gene expression changes associated with U2AF1 mutations, we analyzed the global mRNA expression profile obtained from bone marrow CD34+ cells purified from 5 MDS patients with a U2AF1 mutation, 10 MDS patients without a mutation, and 4 normal donors.
Project description:We investigated the spectra of circulating miRNAs in plasma of myelodysplastic syndromes (MDS) patients. Peripheral blood plasma from MDS patients with different risk scores was used for Agilent miRNA expression microarray analysis to define miRNA profile and to find miRNAs with discriminatory levels for lower risk and higher risk MDS. Results were further validated using droplet digital PCR on a larger cohort, enabling absolute quantification of plasma miRNAs and defining miRNAs with prognostic value for the disease.
Project description:This study includes 2 cohorts of samples. First cohort consists out of BM aspirates from 4 healthy individuals and 12 patients diagnosed with AML, MDS or CMML. Second cohort includes 5 healthy individuals and 17 patients diagnosed with AML, MDS or CMML. MDS and CMML patients were treated with 5-AZA on 6 cycles and samples were obtained before the treatment and 15 days after 1 and 6 rounds of treatment.
Project description:A role for reduced ribosomal protein gene dosage in both DBA and 5q- MDS suggests that other forms of MDS might also involve altered expression of ribosomal protein genes. We used microarrays to interrogate the expression of ribosomal proteins from purified hematopoietic stems cells from patients with low risk myelodysplastic syndrome and age-matched healthy controls. Hematopoietic stem cells were sorted from 8 patients with low risk myelodysplastic syndrome and 11 age-matched healthy controls, and total RNA was hybridized to Affymetrix microarrays.
Project description:We report the biological function of Srsf2 in hematopoiesis in conditional knockout mouse models. Ablation of Srsf2 in the hematopoietic lineage caused embryonic lethality, and Srsf2-deficient fetal liver cells showed significantly enhanced apoptosis and decreased hematopoietic stem/progenitor cells. Induced ablation of Srsf2 in adult Mx1Cre/ Srsf2flox/flox mice upon polyinosinic:polycytidylic acid injection demonstrated a significant decrease in lineage-/Sca+/cKit+ cells in bone marrow. To reveal the functional impact of MDS-associated mutations in SRSF2, we profiled global splicing responses on an MDS-L cell line using RASL-seq, and found that the P95H missense mutation and P95 to R102 in-frame 8 amino-acid deletion caused significant changes in alternative splicing. The affected genes were enriched in cancer development and apoptosis. These findings suggest that intact Srsf2 is essential for the functional integrity of the hematopoietic system, and its mutations are likely key driver events to MDS. MDS-L cells (in triplicate) were transfected by srsf2 shRNA only, or pTRIPZ vectors containing both srsf2 shRNA and srsf2 mutants cDNA including P95H and P95 8 amino acid deletion as well as wild-type construct, followed by Dox induction. Total RNAs were extracted and been analyzed by RASL-seq.