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Callum J C Parr, Shota Katayama, Kenji Miki, Yi Kuang, Yoshinori Yoshida, Asuka Morizane, Jun Takahashi, Shinya Yamanaka, Hirohide Saito
The efficiency of pluripotent stem cell differentiation is highly variable, often resulting in heterogeneous populations that contain undifferentiated cells. Here we developed a sensitive, target-specific, and general method for removing undesired cells before transplantation. MicroRNA-302a-5p (miR-302a) is highly and specifically expressed in human pluripotent stem cells and gradually decreases to basal levels during differentiation. We synthesized a new RNA tool, miR-switch, as a live-cell reporter mRNA for miR-302a activity that can specifically detect human induced pluripotent stem cells (hiPSCs) down to a spiked level of 0...
2016: Scientific Reports
Marco Alexander Poleganov, Sarah Eminli, Tim Beissert, Stephanie Herz, Jung-Il Moon, Johanna Goldmann, Arianne Beyer, Rosario Heck, Isabell Burkhart, Diana Barea Roldan, Özlem Türeci, Kevin Yi, Brad Hamilton, Ugur Sahin
mRNA reprogramming results in the generation of genetically stable induced pluripotent stem (iPS) cells while avoiding the risks of genomic integration. Previously published mRNA reprogramming protocols have proven to be inconsistent and time-consuming and mainly restricted to fibroblasts, thereby demonstrating the need for a simple but reproducible protocol applicable to various cell types. So far there have been no published reports using mRNA to reprogram any cell type derived from human blood. Nonmodified synthetic mRNAs are immunogenic and activate cellular defense mechanisms, which can lead to cell death and inhibit mRNA translation upon repetitive transfection...
November 2015: Human Gene Therapy
Joanna E Kontaraki, Maria E Marketou, Fragiskos I Parthenakis, Spyros Maragkoudakis, Evangelos A Zacharis, Stelios Petousis, George E Kochiadakis, Panos E Vardas
MicroRNAs regulate several aspects of physiological and pathologic cardiac hypertrophy, and they represent promising therapeutic targets in cardiovascular disease. We assessed the expression levels of the microRNAs miR-1, miR-133a, miR-26b, miR-208b, miR-499, and miR-21, in 102 patients with essential hypertension and 30 healthy individuals. All patients underwent two-dimensional echocardiography. MicroRNA expression levels in peripheral blood mononuclear cells were quantified by real-time reverse transcription polymerase chain reaction...
October 2015: Journal of the American Society of Hypertension: JASH
Bharathi Suresh, A Madhan Kumar, Hoe-Su Jeong, Youl-Hee Cho, Suresh Ramakrishna, Kye-Seong Kim
Fanconi anemia (FA) is a recessively inherited multigene disease characterized by congenital defects, progressive bone marrow failure, and heightened cancer susceptibility. Monoubiquitination of the FA pathway member FANCD2 contributes to the repair of replication stalling DNA lesions. However, cellular regulation of FANCD2 monoubiquitination remains poorly understood. In the present study, we identified the miR-302 cluster as a potential regulator of FANCD2 by bioinformatics analysis. MicroRNAs (miRNAs) are the major posttranscriptional regulators of a wide variety of biological processes, and have been implicated in a number of diseases...
October 16, 2015: Biochemical and Biophysical Research Communications
Gary B Fogel, Zoya S Kai, Sahar Zargar, Andrew Hinton, G Adam Jones, Augusta S Wong, Sevan G Ficici, Ana D Lopez, Charles C King
MicroRNAs (miRNAs) are small non-coding RNAs that have emerged as critical regulators of human embryonic stem cell (hESC) pluripotency and differentiation. Despite the wealth of information about the role individual that miRNAs play in these two processes, there has yet to be a large-scale temporal analysis of the dynamics of miRNA expression as hESCs move from pluripotency into defined lineages. In this report, we used Next Generation Sequencing (NGS) to map temporal expression of miRNAs over ten 24-hour intervals as pluripotent cells were differentiated into pancreatic endoderm...
December 15, 2015: Gene
Ketti G Oliveira, Fernanda M Malta, Ana C S S Nastri, Azzo Widman, Paola L Faria, Rúbia A F Santana, Venâncio A F Alves, Flair J Carrilho, João R R Pinho
Hepatitis C virus (HCV) infection affects approximately 3 % of the world population. HCV targets hepatic tissue, and most infected patients develop a chronic infection. Currently, studies have demonstrated an association between HCV-RNA replication and miR-122, the most abundant microRNA in the liver. Our aim was to evaluate liver and serum expression of miR-122 in patients infected with HCV genotypes 1 and 3, and to identify possible associations between miR-122 expression and lipid profiles, HCV viral load, apolipoproteins and liver enzymes...
April 2016: Medical Microbiology and Immunology
Wenwen Deng, Xia Cao, Jingjing Chen, Zhijian Zhang, Qingtong Yu, Yan Wang, Genbao Shao, Jie Zhou, Xiangdong Gao, Jiangnan Yu, Ximing Xu
Induced pluripotent stem cells (iPSCs), resulting from the forced expression of cocktails out of transcription factors, such as Oct4, Sox2, Klf4, and c-Myc (OSKM), has shown tremendous potential in regenerative medicine. Although rapid progress has been made recently in the generation of iPSCs, the safety and efficiency remain key issues for further application. In this work, microRNA 302-367 was employed to substitute the oncogenic Klf4 and c-Myc in the OSKM combination as a safer strategy for successful iPSCs generation...
September 2, 2015: ACS Applied Materials & Interfaces
Ronald J Parchem, Nicole Moore, Jennifer L Fish, Jacqueline G Parchem, Tarcio T Braga, Archana Shenoy, Michael C Oldham, John L R Rubenstein, Richard A Schneider, Robert Blelloch
The evolutionarily conserved miR-302 family of microRNAs is expressed during early mammalian embryonic development. Here, we report that deletion of miR-302a-d in mice results in a fully penetrant late embryonic lethal phenotype. Knockout embryos have an anterior neural tube closure defect associated with a thickened neuroepithelium. The neuroepithelium shows increased progenitor proliferation, decreased cell death, and precocious neuronal differentiation. mRNA profiling at multiple time points during neurulation uncovers a complex pattern of changing targets over time...
August 4, 2015: Cell Reports
Ling Chen, Lingqiang Min, Xuefei Wang, Junjie Zhao, Hua Chen, Jing Qin, Weidong Chen, Zhenbin Shen, Zhaoqing Tang, Qiangjun Gan, Yuanyuan Ruan, Yihong Sun, Xinyu Qin, Jianxin Gu
Gastric cancer remains the third leading cause of cancer-related mortality worldwide, and invasion and metastasis of gastric cancer represent the major reason for its poor prognosis. In this study, we found that loss of the receptor for activated C-kinase 1 (RACK1) promoted the metastasis of gastric cancer by enhancing the autocrine expression of IL8 in vitro and in vivo. microRNA (miRNA; miR) array identified that RACK1 modulated the expression of a series of miRNAs, including the miR-302 cluster, and RACK1 modulated the IL8 expression and tumor invasion through miRNA-302c...
September 15, 2015: Cancer Research
X I Jiang, Yong Luo, Shuli Zhao, Qichao Chen, Chao Jiang, Yutian Dai, Yun Chen, Zhigang Cao
The aim of the present study was to investigate the expression of microRNA (miR)-93, miR-320 and miR-16 and to assess their diagnostic value in diabetic patients with erectile dysfunction (ED). A total of 120 individuals were divided into three groups, which included the diabetics with ED group (ED group), the diabetics without ED group (NED group) and the healthy volunteers group (control group). Each group included 40 individuals. Serum samples were collected and reverse transcription quantitative polymerase chain reaction detection of the three types of miRNA was performed and the sensitivity of ED was analyzed by receiver operating characteristic curves...
July 2015: Experimental and Therapeutic Medicine
Staton L Wade, Lee F Langer, James M Ward, Trevor K Archer
MicroRNAs and chromatin remodeling complexes represent powerful epigenetic mechanisms that regulate the pluripotent state. miR-302 is a strong inducer of pluripotency, which is characterized by a distinct chromatin architecture. This suggests that miR-302 regulates global chromatin structure; however, a direct relationship between miR-302 and chromatin remodelers has not been established. Here, we provide data to show that miR-302 regulates Brg1 chromatin remodeling complex composition in human embryonic stem cells (hESCs) through direct repression of the BAF53a and BAF170 subunits...
October 2015: Stem Cells
Maryam Ghasemi-Kasman, Maryam Hajikaram, Hossein Baharvand, Mohammad Javan
BACKGROUND: The conversion of astrocytes to neuroblasts holds great promise for treatment of neurodegenerative and traumatic brain diseases. METHODOLOGY AND PRINCIPAL FINDINGS: Here we have shown that adult human astrocytes could be reprogrammed to neuroblasts by miR-302/367, both in vivo and in vitro. However, the reprogramming of adult mouse astrocytes to neuroblasts required valproic acid (VPA), a histone deacetylase inhibitor. Following induction of astrocytes toward neurons the expression of pluripotency markers were not detected, which suggested direct cell conversion...
2015: PloS One
Yan Wang, Xingyan Lu, Juan He, Weihong Zhao
INTRODUCTION: Mesenchymal stem cells (MSCs) play a central role in the remediation of cell and tissue damage. Erythropoietin (EPO) may enhance the beneficial influence of MSCs during recovery from tissue and organ injuries. Microvesicles (MVs) released from MSCs contribute to the restoration of kidney damage. We studied the influence of EPO on MVs derived from MSCs, and the protective effects of these factors in subjects with chronic kidney disease (CKD). METHODS: The MVs derived from untreated MSCs (MSC-MVs) or from MSCs incubated in different concentrations of EPO (1, 10, 100, and 500 IU/ml EPO-MVs) were used to treat renal injury of unilateral ureteral obstruction (UUO) in vivo, and transforming growth factor-β1 (TGF-β1)-induced fibrosis in a human renal proximal tubular epithelial (HK2) cell line in vitro...
2015: Stem Cell Research & Therapy
Chul Min Yang, Tomohiro Chiba, Boris Brill, Natalia Delis, Viktoria von Manstein, Vida Vafaizadeh, Thomas Oellerich, Bernd Groner
Cellular transformation is initiated by the activation of oncogenes and a closely associated developmental reprogramming of the epigenetic landscape. Transcription factors, regulators of chromatin states and microRNAs influence cell fates in development and stabilize the phenotypes of normal, differentiated cells and of cancer cells. The miR-302/367 cluster, predominantly expressed in human embryonic stem cells (hESs), can promote the cellular reprogramming of human and mouse cells and contribute to the generation of iPSC...
November 15, 2015: International Journal of Cancer. Journal International du Cancer
Lijuan Sun, Bodu Liu, Zhaoyu Lin, Yandan Yao, Yanyang Chen, Yang Li, Jianing Chen, Dongsheng Yu, Zhangui Tang, Bosheng Wang, Shuguang Zeng, Song Fan, Youyuan Wang, Yilin Li, Erwei Song, Jinsong Li
BACKGROUND: Salivary Adenoid cystic carcinoma (SACC) patients with local invasion and lung metastasis are often resistant to conventional therapy such as operation, chemotherapy and radiotherapy. To explore the underling mechanisms, we studied the roles of miRNA in regulating invasiveness of SACC cells. METHODS: MicroRNA profiling was done in SACC cells with microarray. MiRNA mimics or antisense oligonucleotide was transfected and invasiveness of SACC cells was evaluated by adhesion assay and transwell assay...
2015: Molecular Cancer
Gui-Ming Zhang, Chun-Yang Bao, Fang-Ning Wan, Da-Long Cao, Xiao-Jian Qin, Hai-Liang Zhang, Yao Zhu, Bo Dai, Guo-Hai Shi, Ding-Wei Ye
Micro (mi) RNAs are important regulators involved in various physical and pathological processes, including cancer. The miRNA-302 family has been documented as playing a critical role in carcinogenesis. In this study, we investigated the role of miRNA-302a in prostate cancer (PCa). MiRNA-302a expression was detected in 44 PCa tissues and 10 normal prostate tissues, and their clinicopathological significance was analyzed. Cell proliferation and cell cycle analysis were performed on PCa cells that stably expressed miRNA-302a...
2015: PloS One
Zhonghui Zhang, Yuanfan Hong, Di Xiang, Pei Zhu, Elise Wu, Wen Li, Jeffrey Mosenson, Wen-Shu Wu
miR-302/367 is the most abundant miRNA cluster in human embryonic stem cells (hESCs) and can promote somatic cell reprogramming. However, its role in hESCs remains poorly understood. Here, we studied functional roles of the endogenous miR-302/367 cluster in hESCs by employing specific TALE-based transcriptional repressors. We revealed that miR-302/367 cluster dually regulates hESC cell cycle and apoptosis in dose-dependent manner. Gene profiling and functional studies identified key targets of the miR-302/367 cluster in regulating hESC self-renewal and apoptosis...
April 14, 2015: Stem Cell Reports
Mojtaba Tabrizi, Mitra Khalili, Mohammad Vasei, Nazila Nouraei, Nader Mansour Samaei, Ali Khavanin, Mehrdad Khajehei, Seyed Javad Mowla
BACKGROUND: MicroRNAs are involved in key cellular processes regulating, and their misregulation is linked to cancer. The miR-302-367 cluster is exclusively expressed in embryonic stem and carcinoma cells. This cluster also promotes cell reprogramming and stemness process. In contrast, miR-145 is mostly regarded as a tumor suppressor, where it regulates cellular functions such as cell division, differentiation, and apoptosis. By suppressing the main pluripotency factors (OCT4, SOX2, MYC and KLF4), miR-145 silences the self-renewal program in ESCs...
March 2015: Archives of Iranian Medicine
Yongyan Wu, Fayang Liu, Yingying Liu, Xiaolei Liu, Zhiying Ai, Zekun Guo, Yong Zhang
Wnt/β-catenin signalling plays a prominent role in maintaining self-renewal and pluripotency of mouse embryonic stem cells (mESCs). microRNAs (miRNAs) have critical roles in maintaining pluripotency and directing reprogramming. To investigate the effect of GSK3 inhibitors on miRNA expression, we analysed the miRNA expression profile of J1 mESCs in the absence or presence of CHIR99021 (CHIR) or 6-bromoindirubin-3'-oxime (BIO) by small RNA deep-sequencing. The results demonstrate that CHIR and BIO decrease mature miRNAs of most miRNA species, 90...
2015: Scientific Reports
Mahmoud-Reza Rafiee, Afsaneh Malekzadeh Shafaroudi, Sara Rohban, Hamid Khayatzadeh, Hamid Reza Kalhor, Seyed Javad Mowla
OBJECTIVE: MiR-302-367 is a cluster of polycistronic microRNAs that are exclusively expressed in embryonic stem (ES) cells. The miR-302-367 promoter is functional during embryonic development but is turned off in later stages. Motivated by the cancer stem cell hypothesis, we explored the potential expression of miR-302 in brain tumor cell lines. MATERIALS AND METHODS: In the present experimental study, we have tried to expand our knowledge on the expression pattern and functionality of miR302 cluster by quantifying its expression in a series of glioma (A-172, 1321N1, U87MG) and medulloblastoma (DAOY) cell lines...
2015: Cell Journal
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