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Horizontal transfer of microRNAs: molecular mechanisms and clinical applications

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Protein & Cell 2012年第1期3卷 28-37页

作者： Xi Chen Hongwei Liang Junfeng Zhang ke zen Chen-Yu Zhang Jiangsu Engineering Research Center for microRNA Biology and Biotechnology State Key Laboratory of Pharmaceutical BiotechnologySchool of Life SciencesNanjing UniversityNanjing 210093China

A new class of RNA regulatory genes known as microRNAs(miRNAs)has been found to introduce a whole new layer of gene regulation in *** intensive studies of the past several years have demonstrated that miRNAs are not only found intracellularly,but are also detectable outside cells,including in various body fluids(***,plasma,saliva,urine and milk).This phenomenon raises questions about the biological function of such extracellular *** amounts of extracellular miRNAs are enclosed in small membranous vesicles(***,shedding vesicles and apoptotic bodies)or packaged with RNA-binding proteins(***-density lipoprotein,Argonaute 2 and nucleophosmin 1).These miRNAs may function as secreted signaling molecules to influence the recipient cell ***,secreted extracellular miRNAs may reflect molecular changes in the cells from which they are derived and can therefore potentially serve as diagnostic indicators of *** studies also point to the potential application of siRNA/miRNA delivery as a new therapeutic strategy for treating *** this review,we summarize what is known about the mechanism of miRNA *** addition,we describe the pathophysiological roles of secreted miRNAs and their clinical potential as diagnostic biomarkers and therapeutic *** believe that miRNA transfer between cells will have a significant impact on biological research in the coming years.

关键词： microRNA extracellular microRNA microRNA secretion horizontal transfer microvesicle exosome apoptotic body high-density lipoprotein Argonaute 2 nucleophosmin 1 diagnosis therapy

Nuclear microRNAs and their unconventional role in regulating non-coding RNAs

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Protein & Cell 2013年第5期4卷 325-330页

作者： Hongwei Liang Junfeng Zhang ke zen Chen-Yu Zhang Xi Chen Jiangsu Engineering Research Center for microRNA Biology and Biotechnology State Key Laboratory of Pharmaceutical BiotechnologySchool of Life SciencesNanjing UniversityNanjing 210093China

MicroRNAs (miRNAs) are small non-coding RNAs (ncRNAs) that are involved in post-transcriptional gene regulation. It has long been assumed that miRNAs exert their roles only in the cytoplasm, where they recognize their target protein-coding messenger RNAs (mRNAs), and result in translational repression or target mRNA degradation. Recent studies, however, have revealed that mature miRNAs can also be transported from the cytoplasm to the nucleus and that these nuclear miRNAs can function in an unconventional manner to regulate the biogenesis and functions of ncRNAs (including miRNAs and long ncRNAs), adding a new layer of complexity to our understanding of gene regulation. In this review, we summarize recent literature on the working model of these unconventional miRNAs and speculate on their biological significance. We have every reason to believe that these novel models of miRNA function will become a major research topic in gene regulation in eukaryotes.

关键词： microRNA nuclear microRNA non-coding RNA nucleus Argonaute Exportin Importin

MicroRNA-29a modulates axon branching by targeting doublecortin in primary neurons

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Protein & Cell 2014年第2期5卷 160-169页

作者： Hanqin Li Susu Mao Haitao Wang ke zen Chenyu Zhang Liang Li Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnology State Key Laboratory of Pharmaceutical Biotechnology Nanjing University School of Life Sciences Nanjing 210093 China Department of Pharmacy 306th Hospital of PLA Beijing 100101 China

MicroRNAs （miRNAs） are endogenously expressed small, non-coding transcripts that regulate protein expression. Substantial evidences suggest that miRNAs are enriched in central nervous system, where they are hypothesized to play pivotal roles during neural devel- opment. In the present study, we analyzed miRNAs expression in mice cerebral cortex and hippocampus at different developmental stages and found miR-29a increased dramatically at postnatal stages. In addition, we provided strong evidences that miR-29a is enriched in mature neurons both in vitro and in v/vo. Further investigation demonstrated that the activation of gluta- mate receptors induced endogenous miR-29a level in primary neurons. Moreover, we showed that miR-29a directly regulated its target protein Doublecortin （DCX） expression, which further modulated axon branching in primary culture. Together, our results suggested that miR-29a play an important role in neuronal development of mice cerebrum.

关键词： MiRNAs are approximately 21 nucleotide non-codingtranscripts that are derived from hairpin precursors which

Immune modulatory function of abundant immune-related microRNAs in microvesicles from bovine colostrum

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Protein & Cell 2013年第3期4卷 197-210页

作者： Qi Sun Xi Chen Jianxiong Yu ke zen Chen-Yu Zhang Liang Li Jiangsu Engineering Research Center for microRNA Biology and Biotechnology State Key Laboratory of Pharmaceutical BiotechnologyNanjing University School of Life SciencesNanjing 210093China Department of Virology University of California School of Public HealthBerkeleyCA 94720USA

Colostrum provides essential nutrients and immunologically active factors that are beneficial to *** previous work demonstrated that milk contains large amounts of miRNA that is largely stored in milk-derived microvesicles(MVs).In the present study,we found that the MVs from colostrum contain signifi cantly higher levels of several immune-related *** hypothesized that the colostrum MVs may transfer the immune-related miR-NAs into cells,which contribute to its immune modulatory *** isolated colostrum MVs by ultracentrifugation and demonstrated several immune modulation features associated with *** also provide evidence that the physical structure of milk-derived MVs is essential for transfer miRNAs and following immune modulation ***,we found that colostrum powder-derived MVs also contains higher levels of immune-related miRNAs that display similar immune modulation *** together,these results show that MV-containing immunerelated miRNAs may be a novel mechanism by which co-lostrum modulates body immune response.

关键词： colostrum miRNAs microvesicles immune modulation

MicroRNAs in Drug-induced Liver Injury

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Journal of Clinical and Translational Hepatology 2014年第3期2卷 162-169页

作者： Li-Min Li Dong Wang ke zen State key Laboratory of Pharmaceutical Biotechnology School of Life Sciences Nanjing University Nanjing Jiangsu China

Drug-induced liver injury (DILI) is a leading cause of acute liver failure,and a major reason for the recall of marketed *** of potential liver injury is a challenge for clinical management and preclinical drug safety studies,as well as a great obstacle to the development of new,effective and safe ***,serum levels of alanine and aspartate aminotransferases are the gold standard for evaluating liver ***,these levels are assessed by nonspecific,insensitive,and non-predictive tests,and often result in false-positive ***,there is an urgent need for better DILI biomarkers to guide risk assessment and patient *** discovery of microRNAs (miRNAs) as a new class of gene expression regulators has triggered an explosion of research,particularly on the measurement of miRNAs in various body fluids as biomarkers for many human *** properties of miRNA-based biomarkers,such as tissue specificity and high stability and sensitivity,suggest they could be used as novel,minimally invasive and stable DILI *** the current review,we summarize recent progress concerning the role of miRNAs in diagnosing and monitoring both clinical and preclinical DILI,and discuss the main advantages and challenges of miRNAs as novel DILI biomarkers.

关键词： Biomarker Diagnosis Drug-induced liver injury(DILI) MicroRNA

Selective secretion of microRNA in CNS system

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Protein & Cell 2013年第4期4卷 243-247页

作者： Qipeng Zhang Jie Xu Qun Chen Xi Chen ke zen Chen-Yu Zhang Jiangsu Engineering Research Center for microRNA Biology and Biotechnology State Key Laboratory of Pharmaceutical BiotechnologySchool of Life SciencesNanjing UniversityNanjing 210093China

EXTRACELLULAR miRNAs ARE PRESENT IN VARIOUS BODY FLUIDS MicroRNAs(miRNAs)are a diverse class of endogenous small non-coding RNAs,which posttranscriptionally regulate gene expression by interacting with their binding sites in target ***,several studies have demonstrated that miRNAs are also detectable outside cells,and these miRNAs may be called extracellular *** miRNAs are found in various body fluids,including plasma and serum(Chen et al.,2008),saliva(Park et al.,2009),amniotic fl uid(Weber et al.,2010),Bronchial lavage(Molina-Pinelo et al.,2012;Weber et al.,2010),sputum(Xie et al.,2010),tears and urine(Weber et al.,2010),bile(Shigehara et al.,2011),seminal plasma(Wang et al.,2011),breast milk(Zhou et al.,2012),peritoneal fluid(Chen et al.,2012a).Finding the extracellular miRNAs extended the research field of miRNAs,and got a lot of fruitful research fi ndings.

关键词： al. Weber endogenous

Identification of Ebola virus microRNAs and their putative pathological function

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Science China(Life Sciences) 2014年第10期57卷 973-981页

作者： LIANG HongWei ZHOU Zhen ZHANG SuYang zen ke CHEN Xi ZHANG ChenYu Jiangsu Engineering Research Center for microRNA Biology and Biotechnology State Key Laboratory of Pharmaceutical BiotechnologySchool of Life Sciences Nanjing University

Ebola virus(EBOV),a member of the filovirus family,is an enveloped negative-sense RNA virus that causes lethal infections in humans and ***,more than 1000 people have been killed by the Ebola virus disease in Africa,yet no specific treatment or diagnostic tests for EBOV are *** this study,we identified two putative viral microRNA precursors(pre-miRNAs)and three putative mature microRNAs(miRNAs)derived from the EBOV *** production of the EBOV miRNAs was further validated in HEK293T cells transfected with a pcDNA6.2-GW/EmGFP-EBOV-pre-miRNA plasmid,indicating that EBOV miRNAs can be produced through the cellular miRNA processing *** also predicted the potential target genes of these EBOV miRNAs and their possible biological ***,this study reports for the first time that EBOV may produce miRNAs,which could serve as non-invasive biomarkers for the diagnosis and prognosis of EBOV infection and as therapeutic targets for Ebola viral infection treatment.

关键词： microRNA Ebola virus function

The protective role of myeloid-derived suppressor cells in concanavalin A-induced hepatic injury

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Protein & Cell 2014年第9期5卷 714-724页

作者： Wenli Diao Fangfang Jin Bing Wang Chen-Yu Zhang Jiangning Chen ke zen Limin Li Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnology State Key Laboratory of Pharmaceutical Biotechnology School of Life Sciences Nanjing University Nanjing 210093 China People's Liberation Army 404 Hospital Weihai 264200 China

The mechanism underlying T cell-mediated fulminant hepatitis is not fully understood. In this study, we investigated whether myeloid derived suppressor cells （MDSCs） could prevent the concanavalin A （ConA）- induced hepatitis through suppressing T cell proliferation. We observed an increase in the frequencies of MDSCs in mouse spleen and liver at early stage of ConA treatment, implicating that the MDSCs might be involved in the initial resistance of mice against ConA- mediated inflammation. Subpopulation analysis showed that the MDSCs in liver of ConA-induced mice were mainly granulocytic MDSCs. Adoptive transfer of the bone marrow-derived MDSCs into ConA-treated mice showed that the MDSCs migrated into the liver and spleen where they suppressed T cell proliferation through ROS pathway. In addition, the frequencies of MDSCs in mice were also significantly increased by the treatment with immune suppressor glucocorticoids. Transfer of MDSCs into the regulatory T cell （Treg）- depleted mice showed that the protective effect of MDSCs on ConA-induced hepatitis is Treg-independent. In conclusion, our results demonstrate that MDSCs possess a direct protective role in T cell-mediated hepatitis, and increasing the frequency of MDSCs by either adoptive transfer or glucocorticoid treatment represents a potential cell-based therapeutic strategy for the acute inflammatory disease.

关键词： myeloid derived suppressor cells T cell-mediated hepatitis ROS glucocorticoids concanavalin A(ConA) adoptive transfer glucocorticoid treatment

Accurate quantification of 3'-terminal 2'-O-methylated small RNAs by utilizing oxidative deep sequencing and stem-loop RT-qPCR

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Frontiers of Medicine 2022年第2期16卷 240-250页

作者： Yan Kong Huanhuan Hu Yangyang Shan Zhen Zhou ke zen Yulu Sun Rong Yang Zheng Fu Xi Chen Nanjing Drum Tower Hospital Center of Molecular Diagnostic and Therapy State Key Laboratory of Pharmaceutical BiotechnologyJiangsu Engineering Research Center for MicroRNA Biology and BiotechnologyNJU Advanced Institute of Life Sciences(NAILS)Institute of Artificial Intelligence BiomedicineSchool of Life SciencesNanjing UniversityNanjing 210023China Department of Urology Drum Tower HospitalMedical School of Nanjing UniversityInstitute of UrologyNanjing UniversityNanjing 210008China Department of General Surgery Drum Tower HospitalMedical school of Nanjing UniversityNanjing 210008China Research Unit of Extracellular RNA Chinese Academy of Medical SciencesNanjing 210023China

The continuing discoveries of novel classes of RNA modifications in various organisms have raised the need for improving sensitive,convenient,and reliable methods for quantifying RNA *** particular,a subset of small RNAs,including microRNAs(miRNAs)and Piwi-interacting RNAs(piRNAs),are modified at their 3'-terminal nucleotides via 2'-***,quantifying the levels of these small RNAs is difficult because 2'-0-methylation at the RNA 3'-terminus inhibits the activity of polyadenylate polymerase and T4 RNA *** two enzymes are indispensable for RNA labeling or ligation in conventional miRNA quantification *** this study,we profiled 3'-terminal 2'-0-methyl plant miRNAs in the livers of rice-fed mice by oxidative deep sequencing and detected increasing amounts of plant miRNAs with prolonged oxidation *** further compared the efficiency of stem-loop and poly(A)-tailed RT-qPCR in quantifying plant miRNAs in animal tissues and identified stem-loop RT-qPCR as the only suitable ***,stem-loop RT-qPCR was superior to poly(A)-tailed RT-qPCR in quantifying 3'-terminal 2'-0-methyl piRNAs in human seminal *** summary,this study established a standard procedure for quantifying the levels of 3'-terminal 2'-0-methyl miRNAs in plants and *** measurement of the 3'-terminal 2'-0-methylation of small RNAs has profound implications for understanding their pathophysiologic roles in biological systems.

关键词： small RNAs 2'-0-methylation sequencing RT-qPCR