Unknown,Transcriptomics,Genomics,Proteomics

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Profiling of circulating microRNAs by RT-PCR arrays in atypical hemolytic uremic syndrome and controls samples

ABSTRACT: In this experiment, we explore if circulating microRNAs are altered in patients with atypical hemolytic uremic syndrome (aHUS). To do so, we analyze 4 different pooled samples of aHUS (Pool 1-4) in the arrays and 2 pools of age and gender-matched control samples (Pool F and Pool M). aHUS pools consist of patients samples with different characteristics regarding the presence of complement gene mutations: Pool 1: patients without identified mutations (n=5) Pool 2: patients with mutations in CFH (n=5) Pool 3: patients with mutations in MCP (n=5) Pool 4: patients with mutations in C3 or CFB (n=5) In the case of the control samples, pool F consists of 10 samples of healthy women and pool M consists of 10 samples of healthy men. In conclusion, circulating miRNAs levels are altered in aHUS compared with controls. This alteration is variable and could be mediated by the presence of the complement genetic defect.

ORGANISM(S): Homo sapiens

SUBMITTER: Elena Goicoechea de Jorge

PROVIDER: E-MTAB-8899 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Low fertility remains a leading cause of poor productivity in dairy cattle. In this context, there is significant interest in developing novel tools for accurate early diagnosis of pregnancy. MicroRNAs (miRNAs) are short RNA molecules which are critically involved in regulating gene expression during both health and disease. MiRNAs have been shown to regulate ovarian function, uterine receptivity, embryonic development and placental function. Circulating miRNAs can provide useful biomarkers of tissue function and disease; importantly, differential miRNA profiles have been linked to pregnancy and preeclampsia in humans. This study sought to establish the potential of circulating miRNAs as biomarkers of early pregnancy in cattle. We applied Illumina small-RNA sequencing to profile miRNAs in plasma samples collected from eight non-pregnant heifers on Days 0, 8 and 16 of the oestrous cycle and 11 heifers on Days 16 and 24 of pregnancy. We sequenced a total of 46 samples and generated 9.2 million miRNA reads per sample. There were no differences in miRNA read abundance between any of the pregnant and non-pregnant time-points (FDRâ>â0.1). As a complementary approach, we analysed sample pools (3-4 samples/pool) corresponding to Days 0, 8 and 16 of the oestrous cycle and Day 24 of pregnancy (nâ=â3 pools/group) using Qiagen PCR arrays. A total of 16 miRNAs were differentially expressed (FDRâ<â0.1) in plasma between pregnant and non-pregnant animals. RT-qPCR validation using the same plasma samples confirmed that miR-26a was differentially upregulated on Day 16 pregnant relative to non-pregnant heifers (1.7-fold; Pâ=â0.043), whereas miR-1249 tended to be upregulated in Day 16 pregnant heifers (1.6-fold; Pâ=â0.081). Further validation in an independent group of heifers confirmed an increase in plasma miR-26a levels during early pregnancy, which was significant only on Day 24 (2.0-fold; Pâ=â0.027). Through genome-wide analyses we have successfully profiled plasma miRNA populations associated with early pregnancy in cattle. We have identified miR-26a as a potential circulating biomarker of early pregnancy. Sequencing of three sequencial samples from each of eight cycling animals (Days 0, 8 and 16, total 24 samples) and two samples from each of 11 pregnant animals (Days 16 and 24, total 22 samples). Main comparisons were between non-pregnant and pregnant groups

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Profiling of circulating microRNAs by RT-PCR arrays in atypical hemolytic uremic syndrome and controls samples

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