Project description:To elucidate the function of 30Kc19α-Lin28A protein in osteogenic differentiation of urine-derived stem cells, we established urine-derived stem cell lines differentiated with or without protein treatment. We then performed gene expression profiling analysis using data obtained from RNA-seq of osteogenic differentiated urine-derived stem cells with or without 30Kc19α-Lin28A protein treatment, undifferentiated urine-derived stem cells, and human osteoblasts
Project description:Early diagnosis and treatment is pivotal to the management of kidney disease, whereas the pathological mechanisms and minimally invasive diagnostic method still need to be investigated. In the present study, single-cell RNA sequencing (scRNA-seq) was used to evaluate the heterogeneity of kidney diseases in single cell level. Cellular gene expression profiles of cells of renal tissue, peripheral blood mononuclear cells (PBMCs) and urine from four nephritis patients were performed. Our analysis revealed 12 subsets of renal cells and leukocytes, including fibroblast cells, mesangial cells, epithelial cells, proximal tubule cells (PTCs), and 6 types of immune cells, CD8+ T cell, macrophages (MC), nature killer cells (NK), dendritic cells (DC), B cell and neutrophils. PTCs were detected in both PBMC and urine, while PTC was negative in healthy blood sample. Multiple populations of fibroblast cells, mesangial cells and PTCs demonstrated pro-inflammatory or pro-apoptotic responses. Gene expression analysis suggested that chemotactic and activating effect of inflammatory PTCs and fibroblasts on neutropils were critical for the development and progress of nephritis, which was supported by the widely overexpressed pro-inflammatory genes in these cells. Gene expression profiles of inflammatory PTCs in PBMC, urine and kidney are highly correlated, indicating the high possibility of urine and PBMC PTCs in serving as a surrogate for kidney biopsies.
Project description:Urine is a non-invasive biofluid for the identification of biomarkers to detect disease. In particular extracellular vesicles (EVs) have gained increased interest as a biomarker source, because the molecular content is protected against degradation. Clinical implementation on a daily basis requires protocols that inevitably includes short-term storage of the clinical samples, especially when samples are collected at home. However, little is known about the effect of delayed processing on the urinary EVs and their proteome. In the current study, we evaluated two different storage protocols. First, urine stored at 4˚C without any preservative, and second, a protocol compatible with at-home collection, urine with 40 mM EDTA stored at room temperature. For both conditions it was determined whether storage for 0, 2, 4 and 8 days leads to a change in the global urinary EV proteome profile using proteomics based on data-independent acquisition mass spectrometry. We show that EDTA does not affect the global proteome. Remarkably, the EV proteome was stable in both urine stored up to a week at room temperature with EDTA and in urine stored at 4˚C. These findings open up biomarker studies in urine collected via self-sampling.
Project description:MicroRNA (miRNA) biomarkers for fragile X syndrome were searched by urine microRNA (miRNA) profiling using deep sequencing. The urine miRNA profile of twin boys who shared the same environment but one had a FXS full mutation and the other carried a premutation allele was compared based on the similar sequence reads. The urine of twin boys showed 28 differentiatially regulated miRNAs when 219 reliable identified miRNAs were compared.