Exosome研究成果《MicroRNA profiling of circulating exosomes during experimental liver fibrosis》榮獲2014年美國肝病會(huì)議總統(tǒng)獎(jiǎng)
大家期待已久的101Bio的PureExo Exosome Isolation Kits客戶使用的文章即將見刊。
美國Nationwide Childrens Hospital和The Ohio State University的研究團(tuán)隊(duì)使用美國101Bio的PureExo Exosome Isolation Kits從血清中分離出高純的exosome,并開展了下游的研究工作。共同完成文章《MicroRNA profiling of circulating exosomes during experimental liver fibrosis》。該文章摘要投到2014年的美國肝病會(huì)議(AASLD)后,被授予了總統(tǒng)獎(jiǎng)(Presidential honor),并刊登在這一期的肝臟病雜志(《Hepatology》)上:http://onlinelibrary.wiley.com/doi/10.1002/hep.27500/pdf(請(qǐng)見Abstract #430),正式文章也將在該雜志發(fā)表。
MicroRNA profiling of circulating exosomes during experimental liver fibrosis
Li Chen1, Ruju Chen1, David Brigstock1,2;
1Clinical & Translational Research, Nationwide Childrens Hospital, Columbus, OH; 2Surgery, The Ohio State University, Columbus, OH
Background: Exosomes arise by inward budding of the limiting membranes of multivesicular bodies which, upon fusion with the plasma membrane, result in their secretion and deposition into body fluids (e.g. blood, urine). Exosomes contain a complex mixture of microRNAs (miRs), mRNAs and proteins that reflect the transcriptional and translational status of the producer cell. Since this molecular payload is a “fingerprint” of the dynamic status of their producer cells, exosomes represent a potentially valuable resource for assessing liver disease or pathology. Our goal was to profile the microRNA content of serum exosomes in experimental liver fibrosis. Methods: PureExo Exosome Isolation Kits were used to isolate serum exosomes. MiR profiling was performed on exosomal RNA from 1ml of pooled serum (5 mice; 200μl/mouse) using a mouse miRnome miR PCR Array. miR profiling was performed for the 940 best characterized miRs in the mouse miRnome on exosomes isolated from the circulation of mice after 1 or 5 weeks of treatment with CCl4, as compared to oil-treated controls, with liver injury/fibrosis confirmed histologically. Differentially expressed miRs were confirmed and/or further evaluated by qRT-PCR of exosomal RNA independently obtained at 1-, 4- or 5-weeks of CCl4 administration (n=5). Results: Isolated exosomes from mice serum were bi-membrane vesicles, 50-200nm in diameter, and positive for the exosome markers, CD9 and flotillin-1. Microarray analysis revealed significant alterations in the expression of many hundreds of miRs after either 1- or 5-wks of CCl4 treatment as compared to their respective oil controls. We then focused on selected miRs previously reported to be altered in fibrotic liver, and confirmed the data by RT-PCR. The exosomal levels of these miRs after 5 weeks of CCl4 (including up-regulation of miR-7a, -21, -22, -24, -34a, -155, or -195, and down-regulation of miR-27a, -192, -214, or -377) reflected their previously documented changes at the tissue level in fibrotic liver. In addition, several exosomal miRs that have not yet to be reported in the literature as being altered during liver fibrosis emerged as potentially novel fibrosis markers (e.g. up-regulation of miR-26b or -122; down-regulation of miR-9 or -196b). As compared to their levels at 5 weeks, many of these miRs exhibited individually distinct patterns of expression during the course of fibrosis progression. Conclusions: Dynamic changes occur in the miR content of circulating exosomes during experimental hepatic fibrosis supporting the concept that fibrosis progression and severity is amenable to minimally-invasive assessment through determination of signature exosomal miRs.
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