Dataset Information

Detection of microRNA expression in human peripheral blood microvesicles.

ABSTRACT: MicroRNAs (miRNA) are small non-coding RNAs that regulate translation of mRNA and protein. Loss or enhanced expression of miRNAs is associated with several diseases, including cancer. However, the identification of circulating miRNA in healthy donors is not well characterized. Microvesicles, also known as exosomes or microparticles, circulate in the peripheral blood and can stimulate cellular signaling. In this study, we hypothesized that under normal healthy conditions, microvesicles contain miRNAs, contributing to biological homeostasis.Microvesicles were isolated from the plasma of normal healthy individuals. RNA was isolated from both the microvesicles and matched mononuclear cells and profiled for 420 known mature miRNAs by real-time PCR. Hierarchical clustering of the data sets indicated significant differences in miRNA expression between peripheral blood mononuclear cells (PBMC) and plasma microvesicles. We observed 71 miRNAs co-expressed between microvesicles and PBMC. Notably, we found 33 and 4 significantly differentially expressed miRNAs in the plasma microvesicles and mononuclear cells, respectively. Prediction of the gene targets and associated biological pathways regulated by the detected miRNAs was performed. The majority of the miRNAs expressed in the microvesicles from the blood were predicted to regulate cellular differentiation of blood cells and metabolic pathways. Interestingly, a select few miRNAs were also predicted to be important modulators of immune function.This study is the first to identify and define miRNA expression in circulating plasma microvesicles of normal subjects. The data generated from this study provides a basis for future studies to determine the predictive role of peripheral blood miRNA signatures in human disease and will enable the definition of the biological processes regulated by these miRNA.

SUBMITTER: Hunter MP

PROVIDER: S-EPMC2577891 | biostudies-literature | 2008

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Detection of microRNA expression in human peripheral blood microvesicles.

Hunter Melissa Piper MP Ismail Noura N Zhang Xiaoli X Aguda Baltazar D BD Lee Eun Joo EJ Yu Lianbo L Xiao Tao T Schafer Jeffrey J Lee Mei-Ling Ting ML Schmittgen Thomas D TD Nana-Sinkam S Patrick SP Jarjoura David D Marsh Clay B CB

PloS one 20081111 11

<h4>Background</h4>MicroRNAs (miRNA) are small non-coding RNAs that regulate translation of mRNA and protein. Loss or enhanced expression of miRNAs is associated with several diseases, including cancer. However, the identification of circulating miRNA in healthy donors is not well characterized. Microvesicles, also known as exosomes or microparticles, circulate in the peripheral blood and can stimulate cellular signaling. In this study, we hypothesized that under normal healthy conditions, micro ...[more]

PMID: 19002258

Similar Datasets

Project description:BACKGROUND: MicroRNAs (miRNAs) have been recognized as one of the key regulatory non-coding RNAs that are involved in a number of basic cellular processes. miRNA expression profiling helps to identify miRNAs that could serve as biomarkers. Next generation sequencing (NGS) platforms provide the most effective way of miRNA profiling, particularly as expression of different isoforms of miRNA (IsomiRs) can be estimated by NGS. Therefore, it is now possible to discern the overall complexity of miRNA populations that participate in gene regulatory networks. It is thus important to consider different isoforms of miRNA as part of total profiling in order to understand all aspects of the biology of miRNAs. RESULTS: Here next generation sequencing data of small RNAs derived from normal peripheral blood mononuclear cells (PBMC) and Chronic myeloid leukemia (CML) patients has been used to generate miRNA profiles using a computation pipeline which can identify isomiRs that are natural variants of mature miRNAs. IsomiR profiles have been generated for all the 5p and 3p miRNAs (previously known as major mature miRNA and minor or miRNA*) and the data has been presented as a composite total miRNA transcriptome. The results indicated that the most abundant isomiR sequence of about 68% miRNAs, did not match the reference miRNA sequence as entered in the miRBase and that there is a definite pattern in relative concentration of different isomiRs derived from same precursors. Finally, a total of 17 potential novel miRNA sequences were identified suggesting that there are still some new miRNAs yet to be discovered. CONCLUSIONS: Inclusion of different isoforms provides a detailed miRnome of a cell type or tissues. Availability of miRnome will be useful for finding biomarkers of different cell types and disease states. Our results also indicate that the relative expression levels of different isoforms of a miRNA are likely to be dynamic and may change with respect to changes in the cell or differentiation status.

Project description:Seventh generation Exiqon locked nucleic acid miRCURY LNA microarrays were used to profile the expression of microRNAs in whole blood of patients with lung cancer and of clinically relevant controls without the disease (individuals with non-cancerous lung nodule, at high risk for cancer because of heavy smoking, or with lung cancer treated by surgical resection). The goal was to examine if whole blood microRNAs can be used for developing a non-invasive assay to diagnose lung cancer. Using the PAXgene Blood miRNA kit (Qiagen, Valencia, CA, USA), total RNA was isolated from whole blood stabilized and stored in PAXgene Blood RNA tubes for 85 individuals with lung cancer, 59 individuals without the disease but at a high risk for the disease because of smoking, and 17 individuals with a non-cancerous pulmonary nodule. RNA was also isolated from 13 samples of blood of 12 of the lung cancer cases following surgery; blood samples were collected on two different days post-operatively for one of the cases (Sample names 157 and 160). The data-set here includes data (sample name 171) from a post-operative blood sample of an individual (who had the lung cancer removed by operation) for whom good array data was not obtained for RNA from a pre-operative blood sample. Two RNA samples were used to test technical duplicability of the microarray platform (pairs of sample names 59 & 192, and 175 & 182). Thus, a total of 162 unique individuals and 175 unique RNA samples are covered by this data-set. Isolated RNA was provided to Exiqon (Denmark) for microRNA quantification using microarrays. Quality and concentration of RNA in the samples was assessed by Nanodrop 2000 (Thermo Scientific, Wilmington, DE, USA) and Bioanalyzer 2100 (Agilent, Santa Clara, CA, USA) instruments. Sample RNA (500 ng) was labeled with the Cy3-like Hy3 dye, mixed with 500 ng of a human universal reference RNA (product number AM6000, Ambion, Austin, TX, USA) that was labeled with the Cy5-like Hy5 dye, and hybridized to 7th generation miRCURY locked nucleic acid oligonucleotide microarrays. Sixty-two artificial small RNAs were spiked-in each RNA sample before labeling. disease state: Lung cancer (before surgical resection of lung cancer): 1, 2, 4, 5, 6, 7, 8, 9, 10, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 61, 63, 64, 65, 67, 68, 69, 70, 81, 82, 85, 86, 87, 88, 89, 90, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 121, 122, 123, 124, 125, 126, 127, 149, 151, 153, 154, 156, 158, 165, 167, 174, 175, 176, 177, 179, 180, 181, 183, 184, 186, 187, 188, 189, 193 disease state: Non-cancerous lung nodule: 128, 129, 130, 131, 133, 134, 135, 136, 137, 140, 141, 142, 144, 145, 146, 148, 182, 190 disease state: None (after surgical resection of lung cancer): 155, 157, 159, 160, 161, 162, 163, 164, 166, 169, 170, 171, 172, 173 disease state: None (no current or past lung cancer or non-cancerous nodule): 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 113, 114, 115, 116, 117, 118, 119, 120, 192 individual: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162

Dataset Information

Detection of microRNA expression in human peripheral blood microvesicles.

Publications

Detection of microRNA expression in human peripheral blood microvesicles.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets