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The most common scoring system for critically ill patients is the Sequential Organ Failure Assessment (SOFA) score. Little is known about specific molecular signaling networks underlying the SOFA criteria. We characterized these networks and identified specific key regulatory molecules. We prospectively studied seven patients with sepsis and six controls with high-throughput RNA sequencing (RNAseq). Quantitative reverse transcription PCR (RT-qPCR) confirmation was performed in a second independent cohort. Differentially and significantly expressed miRNAs and their target mRNA transcripts were filtered for admission SOFA criteria and marker RNAs for the respective criteria identified. We bioinformatically constructed molecular signaling networks specifically reflecting these criteria followed by RT-qPCR confirmation of RNAs with important regulatory functions in the networks in the second cohort. RNAseq identified 82 miRNAs (45% upregulated) and 3254 mRNAs (50% upregulated) differentially expressed between sepsis patients and controls. Bioinformatic analysis characterized 6 miRNAs and 76 mRNA target transcripts specific for the SOFA criteria. RT-qPCR validated miRNA and mRNAs included IGFBP2 (respiratory system), MMP9 and PDE4B (nervous system), PPARG (cardiovascular system), AKR1B1, ANXA1, and LNC2/NGAL (acute kidney injury), GFER/ALR (liver), and miR-30c-3p (coagulopathy). There are specific canonical networks underlying the SOFA score. Key regulatory miRNA and mRNA transcripts support its biologic validity.

// Julia Alejandra Pezuk 1, * , Thiago Luiz Araujo Miller 1, 2, * , Jose Luiz Barbosa Bevilacqua 1 , Alfredo Carlos Simoes Dornellas de Barros 1 , Felipe Eduardo Martins de Andrade 1 , Luiza Freire de Andrade e Macedo 1 , Vera Aguilar 1 , Amanda Natasha Menardo Claro 1, 3 , Anamaria Aranha Camargo 1 , Pedro Alexandre Favoretto Galante 1 and Luiz F. L. Reis 1 1 Hospital Sirio-Libanes, Sao Paulo, Brazil 2 Departamento de Bioquimica, Instituto de Quimica, Universidade de Sao Paulo, Sao Paulo, Brazil 3 Current address: SulAmerica, Sao Paulo, Brazil * These authors contributed equally to this work Correspondence to: Pedro Alexandre Favoretto Galante, email: pgalante@mochsl.org.br Luiz F. L. Reis, email: luiz.reis@hsl.org.br Keywords: micro RNAs, breast cancer, BI-RADS, blood plasma, machine learning Received: June 17, 2017 Accepted: July 25, 2017 Published: September 11, 2017 ABSTRACT A BI-RADS category of 4 from a mammogram indicates suspicious breast lesions, which require core biopsies for diagnosis and have an approximately one third chance of being malignant. Human plasma contains many circulating microRNAs, and variations in their circulating levels have been associated with pathologies, including cancer. Here, we present a novel methodology to identify malignant breast lesions in women with BI-RADS 4 mammography. First, we used the miRNome array and qRT-PCR to define circulating microRNAs that were differentially represented in blood samples from women with breast tumor (BI-RADS 5 or 6) in comparison to controls (BI-RADS 1 or 2). Next, we used qRT-PCR to quantify the level of this circulating microRNAs in patients with mammograms presenting with BI-RADS category 4. Finally, we developed a machine learning method (Artificial Neural Network - ANN) that receives circulating microRNA levels and automatically classifies BI-RADS 4 breast lesions as malignant or benign. We identified a minimum set of three circulating miRNAs (miR-15a, miR-101 and miR-144) with altered levels in patients with breast cancer. These three miRNAs were quantified in plasma from 60 patients presenting biopsy-proven BI-RADS 4 lesions. Finally, we constructed a very efficient ANN that could correctly classify BI-RADS 4 lesions as malignant or benign with approximately 92.5% accuracy, 95% specificity and 88% sensibility. We believe that our strategy of using circulating microRNA and a machine learning method to classify BI-RADS 4 breast lesions is a non-invasive, non-stressful and valuable complementary approach to core biopsy in women with BI-RADS 4 lesions.

Graphical abstract Image 1, Prevalence of Kaposi sarcoma-associated herpesvirus (KSHV/HHV-8) varies greatly in different populations. We hypothesized that the actual prevalence of KSHV/HHV8 infection in humans is underestimated by the currently available serological tests. We analyzed four independent patient cohorts with post-surgical or post-chemotherapy sepsis, chronic lymphocytic leukemia and post-surgical patients with abdominal surgical interventions. Levels of specific KSHV-encoded miRNAs were measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and KSHV/HHV-8 IgG were measured by immunoassay. We also measured specific miRNAs from Epstein Barr Virus (EBV), a virus closely related to KSHV/HHV-8, and determined the EBV serological status by ELISA for Epstein-Barr nuclear antigen 1 (EBNA-1) IgG. Finally, we identified the viral miRNAs by in situ hybridization (ISH) in bone marrow cells. In training/validation settings using independent multi-institutional cohorts of 300 plasma samples, we identified in 78.50% of the samples detectable expression of at least one of the three tested KSHV-miRNAs by RT-qPCR, while only 27.57% of samples were found to be seropositive for KSHV/HHV-8 IgG (P, Highlights • There is no agreement on a standard assay to detect the true prevalence of Kaposi sarcoma-associated herpesvirus (KSHV) infection. • Measurement of the viral miRNAs in plasma by RT-qPCR allows a direct and accurate assessment of viral infection. • Measurement of the viral miRNAs in plasma by RT-qPCR shows prevalence of KSHV infection in immuno-depressed patients. • Measurement of plasma viral miRNAs for viral infection assessment has the potential to become a “gold” standard method in the clinical practice. Chronic viral infections represent risk factors for diseases and development of infection-related complications. There is no agreement on a standard assay to detect the true prevalence of Kaposi sarcoma-associated herpesvirus (KSHV) infection. The current method used in the clinical practice (ELISA-test) identifies a great geographic variation in KSHV seroprevalence and may underestimate the true-prevalence of KSHV infection. Here we showed that detection of plasma viral miRNAs levels for the identification of viral infection (e.g., KSHV, Epstein-Bar virus or EBV) is more accurate than the current method for detection of virus-derived antigen, especially in patients with low number of immune cells.

Duchenne muscular dystrophy (DMD) is characterized by progressive skeletal muscle degeneration. No treatments are currently available to prevent the disease. While the muscle enriched microRNA, miR-133b, has been implicated in muscle biogenesis, its role in DMD remains unknown. To assess miR-133b function in DMD-affected skeletal muscles, we genetically ablated miR-133b in the mdx mouse model of DMD. In the absence of miR-133b, the tibialis anterior muscle of P30 mdx mice is smaller in size and exhibits a thickened interstitial space containing more mononucleated cells. Additional analysis revealed that miR-133b deletion influences muscle fiber regeneration, satellite cell proliferation and differentiation, and induces widespread transcriptomic changes in mdx muscle. These include known miR-133b targets as well as genes involved in cell proliferation and fibrosis. Altogether, our data demonstrate that skeletal muscles utilize miR-133b to mitigate the deleterious effects of DMD.

Completion of the human genome sequencing project highlighted the richness of the cellular RNA world, and opened the door to the discovery of a plethora of short and long non-coding RNAs (the dark transcriptome) with regulatory or structural potential, which shifted the balance of pathological gene alterations from coding to non-coding RNAs. Thus, disease risk assessment currently has to also evaluate the expression of new RNAs such as small micro RNAs (miRNAs), long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), competing endogenous RNAs (ceRNAs), retrogressed elements, 3′UTRs of mRNAs, etc. We are interested in the pathogenic mechanisms of atherosclerosis (ATH) progression in patients suffering Chronic Kidney Disease, and in this review, we will focus in the role of the dark transcriptome (non-coding RNAs) in ATH progression. We will focus in miRNAs and in the formation of regulatory axes or networks with their mRNA targets and with the lncRNAs that function as miRNA sponges or competitive inhibitors of miRNA activity. In this sense, we will pay special attention to retrogressed genomic elements, such as processed pseudogenes and Alu repeated elements, that have been recently seen to also function as miRNA sponges, as well as to the use or miRNA derivatives in gene silencing, anti-ATH therapies. Along the review, we will discuss technical developments associated to research in lncRNAs, from sequencing technologies to databases, repositories and algorithms to predict miRNA targets, as well as new approaches to miRNA function, such as integrative or enrichment analysis and their potential to unveil RNA regulatory networks.

[eng] Venous Thrombosis (VT) is a frequent complex disease that involves a disruption of the balance of the hemostatic system. microRNAs (miRNAs) are small non-coding RNAs that regulate gene expression. Promising findings in other complex diseases encourage their use as clinical biomarkers. However, up to now, the implication of miRNAs in VT has not been studied in-depth. The main objective of this Thesis was to study the regulatory mechanisms of the gene expression through miRNAs in VT. The specific objectives were (1) to identify a plasma miRNA profile associated with VT and to analyze its suitability as biomarker, (2) to dissect the interactions between biological layers in the biological context of VT and (3) to identify factors affecting either expression or quantification of miRNAs in plasma. In the first article, the differential expression of miRNAs in plasma was analyzed in the ‘Genetic Analysis of Idiopathic Thrombophilia’ 2 (GAIT-2) Project, which involves 935 individuals belonging to 35 extended Spanish families with idiopathic thrombophilia. First, we conducted a discovery phase, in which 752 miRNAs were measured in plasma by quantitative Polymerase Chain Reaction in 104 individuals of GAIT-2 (52 VT cases and 52 controls). Sixteen miRNAs were selected, which were measured in the entire GAIT-2 (n=935). Four of the miRNAs were significantly associated with VT (false discovery rate 0.7. The first model (VT ~ GATA2 + von Willebrand Factor) showed that the expression of GATA2 in blood was inversely correlated with the blood levels of von Willebrand Factor, and that the disruption of this relationship represents a prothrombotic phenotype. The second model (VT ~ Factor IX, ANXA2 + ENTPD1 + ILK + PDPK1 + PRKAR1A + STXBP3 +hsa-miR- 885-5p + hsa-miR-192-5p) represented an interaction between the fibrinolytic system and platelet activation through the αIIbβ3 signaling pathway. The third model (VT ~CSRP1+ LYN + hsa-miR-192-5p+ hsa-miR-885-5p) revealed the interaction between two group of genes involved in platelet activation, correlated with Protein S, and two miRNAs, in the biological context of VT. In the third article, we used the miRNA expression data of the discovery phase (103 miRNAs in 104 subjects) to analyze the effect of biological and technical factors in their expression. First, we found that the hemolysis marker represented ~10% of the shared variability of miRNA expression. Therefore, using this value as a continuous covariate, beyond as a categorical control, could help to increase consistency across miRNA studies. Second, we found that the expression of miRNAs in plasma was not biased by any blood cell count. Then, we identified 1,323 genetic variants associated with the expression of 16 miRNA genes, that represent 158 independent loci. Finally, we found that these loci were enriched in promoter regions from several tissues, though not in blood tissue. This finding is in agreement with the results regarding blood cell counts, and encourages the role of circulating miRNAs as biomarkers of tissue specific conditions., [spa] El objetivo general de esta Tesis fue estudiar los mecanismos de regulación de la expresión génica mediante microRNAs (miRNAs) en la trombosis venosa (VT). En el primer artículo, se explora la expresión diferencial de miRNAs en VT utilizando la población del proyecto ‘Genetic Analysis of Idiopathic Thrombophilia’ 2 (GAIT-2), compuesta por 935 individuos de 35 familias con trombosis idiopática. El diseño experimental implicó una fase de descubrimiento en la que 752 miRNAs plasmáticos se cuantificaron en 104 individuos del GAIT2 (52 casos y 52 controles) y se seleccionaron 16 miRNAs para la fase de validación, en la que se cuantificaron en toda la población GAIT-2 (n=935). Cuatro de los miRNAs se asociaron significativamente con la enfermedad (false discovery rate

The general goal of this thesis has been to progress oncolytic adenovirus therapy for PDAC, by the incorporation of novel preclinical models to test for patient-specific responses and the generation of oncolytic adenoviruses with enhanced therapeutic index. The two main objectives have been the following: i) Evaluate patients-derived organoids (PDOs) technology as a platform to screen for personalized virotherapy in vitro 1) Establishment of a battery of PDOs from PDAC and normal pancreatic tissues, and evaluation of their applicability in the study of adenoviral infection; 2) Screening of a battery of PDOs to identify individual sensitivities to virotherapies, and the effects derived from the combination with chemotherapy; 3) Study virotherapy-responses in metastasis originated from PDOs xenografted in mice; (ii) Improve oncolytic adenovirus potency by modulation of miRNAs deregulated in PDAC 4) Screening of aberrantly expressed miRNAs sensitizing viral oncolysis in PDAC via CRISPR/Cas9 system; 5) Generation of a miRNA sponge-adenovirus and evaluation of its oncolytic effects in vitro and in vivo; 6) Modulation of miRNA levels with the THZ1 transcriptional inhibitor, and assessment of the effects of its combination with oncolytic adenoviruses.

Full differentiation potential along with self‐renewal capacity is a major property of pluripotent stem cells (PSCs). However, the differentiation capacity frequently decreases during expansion of PSCs in vitro . We show here that transient exposure to a single microRNA, expressed at early stages during normal development, improves the differentiation capacity of already‐established murine and human PSCs. Short exposure to miR‐203 in PSCs (mi PSCs) induces a transient expression of 2C markers that later results in expanded differentiation potency to multiple lineages, as well as improved efficiency in tetraploid complementation and human–mouse interspecies chimerism assays. Mechanistically, these effects are at least partially mediated by direct repression of de novo DNA methyltransferases Dnmt3a and Dnmt3b, leading to transient and reversible erasure of DNA methylation. These data support the use of transient exposure to miR‐203 as a versatile method to reset the epigenetic memory in PSCs, and improve their effectiveness in regenerative medicine.

MicroRNAs from the miR-200 family are commonly associated with the inhibition of the metastatic potential of cancer cells, following inhibition of ZEB transcription factors expression and epithelial-to-mesenchymal transition. However, previous studies performed in pancreatic adenocarcinoma revealed a more complex picture challenging this canonical model. To gain better insights into the role of miR-200 family members in this disease, we analyzed the expression of miR-200a, miR-200b, miR-200c, miR-141, miR-429, and miR-205, and ZEB1, ZEB2, and CDH1 in pancreatic tumors and matching normal adjacent parenchyma and patient-derived xenografts. We found that miR-200a, miR-429, and miR-205 are frequently overexpressed in pancreatic tumors, whereas CDH1 is downregulated, and ZEB1 and ZEB2 levels remain unchanged. Furthermore, we measured a positive correlation between miR-200 family members and CDH1 expression, and a negative correlation between ZEB1 and miR-200c, miR-141, and miR-205 expression, respectively. Interestingly, we identified significant changes in expression of epithelial-to-mesenchymal transition regulators and miR-200 members in patient-derived xenografts. Lastly, functional studies revealed that miR-141 and miR-429 inhibit the tumorigenic potential of pancreatic cancer cells. Taken together, this comprehensive analysis strongly suggests that miRNAs from the miR-200 family, and in particular miR-429, may act as a tumor suppressor gene in pancreatic cancer. Keywords: pancreatic cancer, patient-derived xenograft, miR-200 family, miR-429

[Background] MicroRNAs are small regulatory RNAs with important roles in carcinogenesis. Genetic variants in these regulatory molecules may contribute to disease. We aim to identify allelic variants in microRNAs as susceptibility factors to gastric cancer using association studies and functional approaches., [Methods] Twenty-one single nucleotide variants potentially functional, because of their location in either the seed, mature or precursor region of 22 microRNAs, were selected for association studies. Genetic association with gastric cancer in 365 cases and 1,284 matched controls (European Prospective Investigation into Cancer and Nutrition Cohort) was analysed using logistic regression. MicroRNA overexpression, transcriptome analysis, and target gene validation experiments were performed for functional studies., [Results] rs3746444:T>C, in the seed of MIR499A and mature MIR499B, associated with the cardia adenocarcinoma location; rs12416605:C>T, in the seed of MIR938, associated with the diffuse subtype; and rs2114358:T>C, in the precursor MIR1206, associated with the noncardia phenotype. In all cases, the association was inverse, indicating a protective affect against gastric cancer of the three minor allelic variants. MIR499 rs3746444:T>C and MIR1206 rs2114358:T>C are reported to affect the expression of these miRNAs, but the effect of MIR938 rs12416605:C>T is unknown yet. Functional approaches showed that the expression of MIR938 is affected by rs12416605:C>T and revealed that MIR938 could regulate a subset of cancer-related genes in an allele-specific fashion. Furthermore, we demonstrated that CXCL12, a chemokine participating in gastric cancer metastasis, is specifically regulated by only one of the rs12416605:C>T alleles., [Conclusion] rs12416605 appears to be involved in gastric cancer by affecting the regulatory function of MIR938 on genes related to this cancer type, particularly on CXCL12 posttranscriptional regulation., >la Caixa> Foundation. Grant Number: BM06-130-0; European Regional Development Fund. Grant Number: PI070130 and PI12/01187; Ministerio de Ciencia e Innovación. Grant Number: BFU2010-18477; Fondo de Fomento al Desarrollo Científico y Tecnológico. Grant Number: FONDECYT 1170446; Instituto de Salud Carlos III.

Wenfei Xia,1,* Yun Liu,2,* Yaying Du,1 Teng Cheng,1 Xiaopeng Hu,1 Xingrui Li1 1Department of Breast and Thyroid Surgery, Division of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, People’s Republic of China; 2Department of ENT, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xingrui LiDepartment of Breast and Thyroid Surgery, Division of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 095 Jiefang Avenue, Wuhan, Hubei 430030, People’s Republic of ChinaEmail lixingrui0698@163.comBackground/Aims: The effects of microRNA-423 on proliferation and drug resistance of breast cancer cells were explored, the downstream target genes of miR-423 and the targeted regulatory relationship between them were studied.Methods: RT-qPCR was used to detect the expression of miR-423 in breast cancer tissues and cell lines, and the transfection efficiency of miR-423 inhibitory vector miR-423-inhibitor was constructed and verified. CCK-8 and colony formation assays were used to examine the effect of miR-423 on tumor cell proliferation. Target gene prediction and screening and luciferase reporter assay were used to verify downstream target genes of miR-432. The mRNA and protein expression of miR-423target gene ZFP36 was detected by RT-qPCR and Western blotting.Results: The expression of miR-423 was significantly higher than that in normal tissues. Compared to the non-malignant mammary epithelial cell line MCF-10A, the expression of miR-423 was significantly raised in MCR-7 and MCF-7/ADR cells. ZFP36 was a downstream target gene of miR-423 and negatively correlated with the expression of miR-423 in breast cancer. The knockdown of miR-423 can significantly enhance the cytotoxicity of the drug, increase the apoptotic rate of MCF-7/ADR cells. miR-423 was capable of activating the Wnt/β-catenin signaling pathway leading to chemoresistance and proliferation, whereas overexpression of ZFP36 reduced drug resistance and proliferation.Conclusion: miR-423 acted as an oncogene to promote tumor cell proliferation and migration. ZFP36 was a downstream target gene of miR-423, and miR-423 inhibited the expression of ZFP36 via Wnt/β-catenin signaling pathway of breast cancer cells.Keywords: micro RNAs, miR-423, ZFP36, Wnt/β-catenin, breast cancer

Se desarrolló un modelo de diabetes mellitus gestacional (grupo GDM) mediante programación intrauterina en la cría de ratas con diabetes moderada. Los animales gestantes del grupo GDM presentaron, además de hiperglucemia, un aumento en los niveles plasmáticos de insulina y de triglicéridos en relación con el grupo control. Los fetos provenientes del grupo GDM también presentaron hiperglucemia e hiperinsulinemia, junto a un incremento del peso fetal, respecto al grupo control. Se estudió el hígado de fetos de 21 días de preñez (gestación a término). Los fetos del grupo GDM presentaron alteraciones sexo-dependientes en los niveles de lípidos hepáticos y en la expresión de los receptores activados por proliferadores peroxisomales (PPAR): PPARγ y PPARδ, que se acompañaron con alteraciones en la expresión de enzimas blanco de los PPAR involucradas en el anabolismo y/o catabolismo de lípidos. Se evaluó el efecto del tratamiento dietario materno con un suplemento de aceite de oliva durante la gestación en las ratas que desarrollarían GDM. Este tratamiento no logró prevenir la inducción de la diabetes gestacional, pero disminuyó los niveles de triglicéridos plasmáticos maternos y redujo el peso fetal a valores similares al control. A su vez, el tratamiento dietario materno en ratas con GDM reguló la acumulación lipídica y redujo algunos parámetros pro-oxidantes/pro-inflamatorios que se encontraban elevados en el hígado fetal. Este tratamiento, también redujo los elevados niveles de PPARγ y PPARδ en el hígado fetal del grupo GDM, como así también la expresión de sus enzimas blanco que regulan el metabolismo lipídico. Esta reducción se vinculó a una menor expresión de dos de los co-activadores de los PPAR, PGC-1α y SRC-1. Otro aspecto de interés fue estudiar la posible regulación epigenética de los PPAR. Se estudió la expresión de los microRNAs, miR-130a y miR-9, reguladores de PPARγ y PPARδ respectivamente. Evidenciamos de manera sexo-dependiente una reducción en la expresión de ambos microRNAs, los cuales estarían asociados a los elevados niveles de PPARγ y PPARδ, y a la consecuente alteración de los niveles de lípidos en el hígado de fetos del grupo GDM. Además, se estudiaron los niveles del miR-122, el microRNA con mayor expresión en el hígado, implicado en varios aspectos importantes de la fisiopatología hepática. Los niveles del miR-122 se encontraron alterados, en forma sexo-dependiente, en el hígado fetal del grupo GDM, evidenciándose a su vez que la expresión de este microRNA estaría regulada por la actividad del receptor nuclear PPARγ. En conjunto, estos resultados sugieren que las alteraciones en el metabolismo lipídico hepático fetal observadas en el grupo GDM estarían asociadas a una desregulación en la función de los PPAR. El tratamiento dietario materno con aceite de oliva fue capaz de regular la expresión de los PPAR y de las enzimas hepáticas fetales involucradas en el metabolismo lipídico, probablemente mejorando la función hepática en la vida adulta de la descendencia. An experimental model of gestational diabetes mellitus (GDM group) was developed by intrauterine programming in the offspring of mild diabetic rats. Pregnant rats in the GDM group showed not only hyperglycemia, but also hyperinsulinemia and hypertriglyceridemia. The fetuses from the GDM group also showed hyperglycemia and hyperinsulinemia, and had increased weight when compared to controls. The fetal liver was studied on the day 21 of gestation, corresponding to term pregnancy. The fetuses of the GDM group showed sex-dependent alterations in the levels of hepatic lipids and in the expression of peroxisome proliferator activated receptors (PPARs): PPARγ and PPARδ. These anomalies were associated with alterations in the expression of PPAR target enzymes involved in lipid anabolism and/or catabolism. A maternal treatment with olive oil during the gestation of the rats that would develop GDM was evaluated. This dietary treatment was not able to prevent the development of GDM, but led to a decrease in the levels of triglycerides in maternal plasma and in the fetal weight to values similar to those of the controls. Moreover, the maternal treatment with olive oil in the GDM group regulated the lipid levels in the fetal liver and reduced some of the pro-oxidant/pro-inflammatory parameters that were increased in the fetal liver. Also, the maternal treatment with olive oil reduced the expression of PPARγ and PPARδ in the fetal liver in the GDM group, as well as the expression of enzymes that regulate the lipid metabolism and are targets of PPARs. This reduction was related to the reduced levels of PGC-1α and SRC-1, coactivators of the nuclear receptors PPAR. We were also interested in addressing a putative epigenetic regulation of PPARs. Thus, we analyzed the expression of two microRNAs, miR-130a and miR-9, negative regulators of PPARγ and PPARδ in the fetal liver, respectively. We observed a sexdependent reduction in both microRNAs, alterations possibly related to the increased levels of PPARγ and PPARδ and the consequent alterations in lipid levels induced in the fetal liver of the GDM group. In addition, we studied the levels of miR-122, the microRNA with greater expression in the liver, involved in several important aspects of the physiopathology of this organ. We found sex-dependent alterations in the levels of miR-122 in the fetal liver of the GDM group, and also found that the expression of this microRNA can be regulated by the activity of the nuclear receptor PPARγ. Altogether, these results suggest that the alterations of the lipid metabolism in the fetal liver from the GDM group are related to a deregulation in PPARs function. The maternal dietary treatment with olive oil was able to regulate the expression of PPARs and the hepatic fetal enzymes involved in the lipid metabolism in the fetal liver of the GDM group, probably improving the hepatic function in the adult life of the offspring. Fil: Fornés, Daiana Débora. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.

Hypoxia-Inducible Factors (HIFs) play essential roles in the physiological response to low oxygen in all multicellular organisms, while their deregulation is associated with human diseases. HIF levels and activity are primarily controlled by the availability of the oxygen-sensitive HIFα subunits, which is mediated by rapid alterations to the rates of HIFα protein production and degradation. While the pathways that control HIFα degradation are understood in great detail, much less is known about the targeted control of HIFα protein synthesis and what role this has in controlling HIF activity during the hypoxic response. This review will focus on the signalling pathways and RNA binding proteins that modulate HIFα mRNA half-life and/or translation rate, and their contribution to hypoxia-associated diseases.

BACKGROUND: Cancer-derived exosomes are involved in metastasis. YKT6 is a SNARE protein that participates in the regulation of exosome production and release, but its role in non-small cell lung cancer (NSCLC) has not been examined. MATERIALS AND METHODS: Ultracentrifugation-purified exosomes from the A549 cell line were studied by CRYO-TEM, nanoparticle tracking analysis and western blot (TSG101 marker). YKT6 was inhibited using a DsiRNA and selected pre-microRNAs. MicroRNAs targeting YKT6 were validated by Renilla/Luciferase assay and western blot. YKT6 expression and its prognostic impact were analyzed in 98 tissue specimens from resected NSCLC patients. RESULTS: Membranous nanosized vesicles (mode size: 128nm) with TSG101 protein were purified from A549 cells. YKT6 inhibition reduced exosome release by 80.9%. We validated miR-134 and miR-135b as miRNAs targeting YKT6, and transfection with the pre-miRNAs also produced a significant reduction in exosome release. The analysis of YKT6 in tumor samples showed that patients with high levels had shorter disease-free and overall survival. CONCLUSIONS: YKT6 is a key molecule in the regulation of exosome release in lung cancer cells and is in turn precisely regulated by miR-134 and miR-135b. Moreover, YKT6 levels impact prognosis of resected NSCLC patients.

One largely unknown question in cell biology is the discrimination between inconsequential and functional transcriptional events with relevant regulatory functions. Here, we find that the oncofetal HMGA2 gene is aberrantly reexpressed in many tumor types together with its antisense transcribed pseudogene RPSAP52. RPSAP52 is abundantly present in the cytoplasm, where it interacts with the RNA binding protein IGF2BP2/IMP2, facilitating its binding to mRNA targets, promoting their translation by mediating their recruitment on polysomes and enhancing proliferative and self-renewal pathways. Notably, downregulation of RPSAP52 impairs the balance between the oncogene LIN28B and the tumor suppressor let-7 family of miRNAs, inhibits cellular proliferation and migration in vitro and slows down tumor growth in vivo. In addition, high levels of RPSAP52 in patient samples associate with a worse prognosis in sarcomas. Overall, we reveal the roles of a transcribed pseudogene that may display properties of an oncofetal master regulator in human cancers.

MicroRNA-210-3p is the most prominent hypoxia regulated microRNA, and it has been found significantly overexpressed in different human cancers. We performed the expression analysis of miR-210-3p in a retrospective cohort of breast cancer patients with a median follow-up of 76 months (n = 283). An association between higher levels of miR-210-3p and risk of disease progression (HR: 2.13, 95%CI: 1.33-3.39, P = 0.002) was found in the subgroup of patients treated with Epirubicin and Cyclophosphamide followed by Docetaxel. Moreover, a cut off value of 20.966 established by ROC curve analyses allowed to discriminate patients who developed distant metastases with an accuracy of 85% at 3- (AUC: 0.870, 95%CI: 0.690-1.000) and 83% at 5-years follow up (AUC: 0.832, 95%CI: 0.656–1.000). Whereas the accuracy in discriminating patients who died for the disease was of 79.6% at both 5- (AUC: 0.804, 95%CI: 0.517–1.000) and 10-years (AUC: 0.804. 95%CI: 0.517–1.000) follow-up. In silico analysis of miR-210-3p and Docetaxel targets provided evidence for a putative molecular cross-talk involving microtubule regulation, drug efflux metabolism and oxidative stress response. Overall, our data point to the miR-210-3p involvement in the response to therapeutic regimens including Docetaxel in sequential therapy with anthracyclines, suggesting it may represent a predictive biomarker in breast cancer patients.

The study of small RNAs provides us with a deeper understanding of the complexity of gene regulation within cells. Of the different types of small RNAs, the most important in mammals are miRNA, tRNA fragments and piRNAs. Using small RNA-seq analysis, we can study all small RNA types simultaneously, with the potential to detect novel small RNA types. We describe SeqclusterViz, an interactive HTML-javascript webpage for visualizing small noncoding RNAs (small RNAs) detected by Seqcluster. The SeqclusterViz tool allows users to visualize known and novel small RNA types in model or non-model organisms, and to select small RNA candidates for further validation. SeqclusterViz is divided into three panels: i) query-ready tables showing detected small RNA clusters and their genomic locations, ii) the expression profile over the precursor for all the samples together with RNA secondary structures, and iii) the mostly highly expressed sequences. Here, we show the capabilities of the visualization tool and its validation using human brain samples from patients with Parkinson’s disease .

Ovarian cancer is the most lethal gynecological malignancy due to the silent nature on its early onset and the rapid acquisition of drug resistance. Histologically heterogeneous, it includes several subtypes with different mutational landscapes, hampering the development of effective targeted therapies. Non-coding RNAs are emerging as potential new therapeutic targets in cancer. To search for a microRNA signature related to ovarian carcinomas and study its potential as effective targeted therapy, we examined the expression of 768 miRNA in a large collection of tumor samples and found miR-654-5p to be infraexpressed in ovarian serous carcinomas, the most common and aggressive type. Restoration of miR-654-5p levels reduced tumor cell viability in vitro and in vivo and impaired sphere formation capacity and viability of ovarian cancer patient-derived ascitic cells ex vivo. CDCP1 and PLAGL2 oncogenes were found to be the most relevant direct miR-654-5p targets and both genes convey in a molecular signature associated with key cancer pathways relevant to ovarian tumorigenesis, such as MYC, WNT and AKT pathways. Together, we unveiled the tumor suppressor function of miR-654-5p, suggesting that its restoration or co-targeting of CDCP1 and PLAGL2 may be an effective therapeutic approach for ovarian cancer., This work was supported in part by grants from Instituto de la Mujer Dexeus (DEXEUS-B29/012), CIBER (CB16/12/00328), SGR (2017 SGR 1661), the Ministerio de Economia y Competitividad and Fondos FEDER (RTC-2015-3821-1), Instituto Carlos III (PI15/00238 to A.S. and PI17/00564 to M.F.S) and the Miguel Servet Program (CP13/00158 and CPII18/00027 to AS. and CPII16/00006 to MFS). AP and LS were supported by predoctoral VHIR fellowships and CJ by an AGAUR predoctoral fellowship (VHIR: PRED-VHIR-2014-11 and PRED-VHIR-2017; AGAUR: 2017FI_B_00095, respectively).

OBJECTIVES: Pancreatic ductal adenocarcinoma (PDAC) presents the lowest survival rate of all cancers because only 6% of patients reach five-year survival. Alterations in the expression of several microRNAs (miRNAs) occur in the tumor of PDAC and in preneoplastic lesions as the called intraductal papillary mucinous neoplasm (IPMN). Here, we aimed at identifying which miRNAs are significantly altered in liquid biopsies from patients with PDAC and IPMN to find new noninvasive biomarkers for early detection of PDAC. METHODS: We analyzed by real-time quantitative reverse transcription-PCR (qRT-PCR) the expression of 17 circulating miRNAs, previously found to be significantly overexpressed in tissue pancreatic neoplasms, in a set of 182 plasma samples (94 PDAC, 19 IPMN, 18 chronic pancreatitis, and 51 disease-free controls). Then, we analyzed CA19.9 levels in the same plasma set, and we assessed the diagnostic values of differentially expressed miRNAs, CA19.9, and all possible combinations. RESULTS: Of note, 16, 14, and 9 miRNAs were significantly increased in PDAC, IPMN, and chronic pancreatitis, respectively, compared with control plasmas. miR-21-5p, miR-33a-3p, miR-320a, and miR-93-5p showed the highest discriminating capacity for pancreatic neoplasia (PDAC or IPMN) with an area under the receiver operating characteristic curve (AUC) of 0.86, 0.85, 0.85, and 0.80, respectively. 2-miRNA combinations improved these performances reaching AUC = 0.90 for “miR-33a-3p+miR-320a.” Addition of CA19.9 increased the diagnostic potential of miRNA signatures even further achieving an AUC of 0.95 (93% sensitivity and 85% specificity) for the combination of “miR-33a-3p+miR-320a+CA19.9.” CONCLUSIONS: Novel signatures combining miRNAs and CA19.9 could be used as noninvasive biomarkers for early detection of PDAC.

MiRNA (or microRNA) is a subclass of non-coding RNAs that is responsible for post-transcriptional gene regulation. It has approximately 22 nucleotides and regulates gene expression in plants and animals at the post-transcriptional level, by the cleavage of a target mRNA or by suppression of its translation. Although many of the processes and mechanisms have not yet been fully elucidated, there is a strong association between miRNA expression and several diseases. It is known that miRNAs are expressed in the cardiovascular system, but their role in cardiovascular diseases (CVDs) has not been clearly established. In this non-systematic review of the literature, we first present the definition of miRNAs and their action at the cellular level. Afterward, we discuss the role of miRNAs as circulating biomarkers of CVDs, and then their role in cardiac remodeling and atherosclerosis. Despite the complexity and challenges, it is crucial to identify deregulated miRNAs in CVDs, as it allows a better understanding of underlying cellular and molecular mechanisms and helps in the development of more accurate diagnostic and prognostic circulating biomarkers, and new therapeutic strategies for different stages of CVDs. Resumo O miRNA (ou microRNA) constitui uma subclasse de RNAs não codificantes responsáveis pela regulação gênica pós-transcricional. Ele possui aproximadamente 22 nucleotídeos e regula a expressão gênica em plantas e animais ao nível pós-transcricional, pela clivagem de um mRNA alvo ou da repressão de sua tradução. Embora muitos processos e mecanismos ainda não estejam completamente elucidados, existe uma forte associação entre a expressão de miRNAs e diversas doenças que acometem o organismo. Os miRNAs são expressos no sistema cardiovascular, contudo o seu papel no desenvolvimento das doenças cardiovasculares (DCVs) ainda não está totalmente elucidado. Diante disso, realizou-se uma revisão não sistemática da literatura a fim de se discutir a relação entre os miRNAs e as DCVs. Nesta revisão, primeiramente é discutido o que são os miRNAs e a sua ação a nível celular. Após, é discutido o papel dos miRNAs como biomarcadores circulantes de DCVs e então o seu papel no remodelamento cardíaco e na aterosclerose. Apesar da complexidade e dos desafios, a identificação dos miRNAs desregulados nas DCVs é crucial, uma vez que possibilita uma melhor compressão dos mecanismos celulares e moleculares envolvidos, assim como auxilia o desenvolvimento de marcadores circulantes de diagnóstico e prognóstico mais acurados e de novas estratégias terapêuticas para os diferentes estágios da DCV.

Fluoxetine (FLX) is a pharmaceutical used to treat affective disorders in humans, but as environmental contaminant also affects inadvertently exposed fish in urban watersheds. In humans and fish, acute FLX treatment and exposure are linked to endocrine disruption, including effects on the reproductive and stress axes. Using the zebrafish model, we build on the recent finding that developmental FLX exposure reduced cortisol production across generations, to determine possible parental and/or life-stage-dependent (age and/or breeding experience) contributions to this phenotype. Specifically, we combined control and developmentally FLX-exposed animals of both sexes (F 0 ) into four distinct breeding groups mated at 5 and 9 months, and measured offspring (F 1 ) basal cortisol at 12 dpf. Basal cortisol was lower in F 1 descended from developmentally FLX-exposed F 0 females bred at 5, but not 9 months, revealing a maternal, life-stage dependent effect. To investigate potential molecular contributions to this phenotype, we profiled maternally deposited transcripts involved in endocrine stress axis development and regulation, epigenetic (de novo DNA methyltransferases) and post-transcriptional (miRNA pathway components and specific miRNAs) regulation of gene expression in unfertilized eggs. Maternal FLX exposure resulted in decreased transcript abundance of glucocorticoid receptor, dnmt3 paralogues and miRNA pathway components in eggs collected at 5 months, and increased transcript abundance of miRNA pathway components at 9 months. Specific miRNAs predicted to target stress axis transcripts decreased (miR-740) or increased (miR-26, miR-30d, miR-92a, miR-103) in eggs collected from FLX females at 5 months. Increased abundance of miRNA-30d and miRNA-92a persisted in eggs collected from FLX females at 9 months. Clustering and principal component analyses of egg transcript profiles separated eggs collected from FLX-females at 5 months from other groups, suggesting that oocyte molecular signatures, and miRNAs in particular, may serve as predictive tools for the offspring phenotype of reduced basal cortisol in response to maternal FLX exposure. © 2019 Martinez et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited., RM was supported also by a FPU predoctoral fellowship (www.ciencia.gob.es), grant #FPU15/03332, and research mobility grants from the Spanish Ministry of Education, Culture and Sport (www.mecd.gob.es), grant #EST16/00695 and grant #EST17/00830, respectively. This work was supported by grants from the Natural Sciences and Engineering Research Council of Canada (www.nserc-crsng.gc.ca) under the Discovery Grant program, grant #2114456-2017, and The Canada Foundation of Innovation (www.innovation. ca), under the John R. Evans Leaders Fund (https://www.innovation.ca) program, grant #148035, awarded to JAM. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

目的 分析microRNA-138（miR-138）靶向调节小谷氨酰胺三角四肽重复区蛋白α（SGTA）对非霍奇金淋巴瘤（NHL）细胞黏附介导的耐药（cell adhesion-mediated drug resistance, CAM-DR）的影响。 方法 以HS-5细胞或纤连蛋白（FN）构建NHL细胞黏附模型；Western blotting与RQ-PCR分析miR-138对SGTA蛋白及mRNA表达的影响；双荧光素酶报告基因活性检测分析miR-138对SGTA mRNA 3′UTR活性的影响；分别用miR-138下调慢病毒载体（Lv-shmiR-138）、miR-138过表达慢病毒载体（Lv-miR-138）、对照慢病毒载体（Lv-Ctrl）感染Daudi及OCI-Ly8细胞，分析miR-138对细胞周期、黏附能力及CAM-DR的影响；收集35例弥漫大B细胞淋巴瘤患者的石蜡组织标本，分析miR-138的表达情况及其与疾病进展和耐药关系。 结果 ①NHL细胞与FN或HS-5细胞黏附培养后miR-138表达水平显著高于悬浮培养组（P值均0.05）。③转染野生型SGTA 3′UTR载体时，过表达miR-138可显著抑制荧光素酶报告基因活性（0.73±0.03对1.00±0.02，t=0.914，P=0.002）；而转染结合位点突变的突变型载体时，过表达miR-138不能显著改变报告基因活性（0.93±0.04对1.00±0.02，t=1.375，P=0.241）。④在悬浮及黏附培养状态下，过表达miR-138可显著诱导Daudi及OCI-Ly8细胞G1期停滞（P值均0.05）。⑥在悬浮培养状态下调miR-138的表达时，多柔比星诱导细胞死亡的比例显著下降，而在黏附培养状态下调miR-138的表达，多柔比星诱导细胞死亡的比例显著上升（P值均

Background Stem cell therapy is considered as a promising alternative to treat intervertebral disc degeneration (IDD). Extensive work had been done on identifying and comparing different types of candidate stem cells, both in vivo and in vitro. However, few studies have shed light on degenerative nucleus pulposus cells (NPCs), especially their biological behavior under the influence of exogenous stem cells, specifically the gene expression and regulation pattern. In the present study, we aimed to determine messenger RNAs (mRNAs) and long non-coding RNAs (lncRNAs), which are differentially expressed during the co-culturing process with adipose-derived mesenchymal stem cells (ASCs) and to explore the involved signaling pathways and the regulatory networks. Methods We compared degenerative NPCs co-cultured with ASCs with those cultured solely using lncRNA-mRNA microarray analysis. Based on these data, we investigated the significantly regulated signaling pathways based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database. Moreover, 23 micro RNAs (miRNAs), which were demonstrated to be involved in IDD were chosen; we investigated their theoretic regulatory importance associated with our microarray data. Results We found 632 lncRNAs and 1682 mRNAs were differentially expressed out of a total of 40,716 probes. We then confirmed the microarray data by real-time PCR. Furthermore, we demonstrated 197 upregulated, and 373 downregulated Gene Ontology terms and 176 significantly enriched pathways, such as the mitogen-activated protein kinase (MAPK) pathway. Also, a signal-net was constructed to reveal the interplay among differentially expressed genes. Meanwhile, a mRNA-lncRNA co-expression network was constructed for the significantly changed mRNAs and lncRNAs. Also, the competing endogenous RNA (ceRNA) network was built. Conclusion Our results present the first comprehensive identification of differentially expressed lncRNAs and mRNAs of degenerative NPCs, altered by co-culturing with ASCs, and outline the gene expression regulation pattern. These may provide valuable information for better understanding of stem cell therapy and potential candidate biomarkers for IDD treatment. Electronic supplementary material The online version of this article (10.1186/s13075-018-1677-x) contains supplementary material, which is available to authorized users.

microRNAs (miRNAs) play key roles in cancer, but their propensity to couple their targets as competing endogenous RNAs (ceRNAs) has only recently emerged. Multiple models have studied ceRNA regulation, but these models did not account for the effects of co-regulation by miRNAs with many targets. We modeled ceRNA and simulated its effects using established parameters for miRNA/mRNA interaction kinetics while accounting for co-regulation by multiple miRNAs with many targets. Our simulations suggested that co-regulation by many miRNA species is more likely to produce physiologically relevant context-independent couplings. To test this, we studied the overlap of inferred ceRNA networks from four tumor contexts—our proposed pan-cancer ceRNA interactome (PCI). PCI was composed of interactions between genes that were co-regulated by nearly three-times as many miRNAs as other inferred ceRNA interactions. Evidence from expression-profiling datasets suggested that PCI interactions are predictive of gene expression in 12 independent tumor- and non-tumor contexts. Biochemical assays confirmed ceRNA couplings for two PCI subnetworks, including oncogenes CCND1, HIF1A and HMGA2, and tumor suppressors PTEN, RB1 and TP53. Our results suggest that PCI is enriched for context-independent interactions that are coupled by many miRNA species and are more likely to be context independent.

In this paper, an on-line solid-phase extraction capillary electrophoresis mass spectrometry (SPE-CE-MS) method is described for the purification, preconcentration, separation, and characterization of endogenous microRNA (miRNA) and their post-transcriptional modifications in serum. First, analysis by CE-MS was optimized using a standard mixture of hsa-miR-21-5p (miR-21-Sp) and hsa-let-7g-Sp (let-7g-Sp). For SPE-CE-MS, a commercial silicon carbide (SiC) resin was used to prepare the microcartridges. Under the optimized conditions with standards, the micro cartridge lifetime (>25 analyses) and repeatability (2.8% RSD for the migration times; 4.4 and 6.4% RSD for the miR-21-5p and let-7g-5p peak areas, respectively) were good, the method was linear between 25 and 100 nmol.L-1, and the limit of detection (LOD) was around 10 nmol.L-1 (50 times lower than by CE-MS). In order to analyze human serum samples, an offline sample pretreatment based on phenol/chloroform/isoamyl alcohol (PCA) extraction was necessary prior to SPE-CE-MS. The potential of the SPE-CE-MS method to screen for B-cell chronic lymphocytic leukemia (CLL) was demonstrated by an analysis of serum samples from healthy controls and patients. MicroRNAs, specifically miR-21-Sp and a 23 nucleotide long 5'-phosphorylated miRNA with 3'-uridylation (iso-miR-16-5p), were only detected in the CLL patients.

ECRIS Network integrated in the Spanish AIDS Research Network., Exosome-derived miR-21 was independently associated with CD4 T cell decline in HIV-1-infected elite controllers (OR 0.369, 95% CI 0.137–0.994, p = 0.049). Also, a negative correlation between miR-21 expression and MCP-1 level was found (r = −0.649, p = 0.020), while no correlation between soluble biomarkers or cellular immune activation was found., This work has been partially funded by the Spanish Health Institute Carlos III (ISCIII) and European Regional Development Fund, Spanish AIDS Research Network (RIS) with grant numbers RD16/0025/0001, RD12/0017/0031, RD16/0025/0013, and RD16/0002/0002. Research Grant Project, Spanish Health Research Fund (FIS, ISCIII) number PI14/00011. Miguel Servet Research Program, Spanish Health Institute Carlos III (ISCIII) CP14/00198 to NR. Spanish Health Research Fund (FIS, ISCIII) and IIS-FJD Intramural Research Scholarship co-funded grant number CP14/00198 to MG. Spanish Health Institute Carlos III (ISCIII) grant CD13/00013 to MAJS. Grant Research, Madrid Education, Youth and Sport Counseling number PEJ15/bio/tl-0064 to MJRL.

Circulating small RNAs, including miRNAs but also isomiRs and other RNA species, have the potential to be used as non-invasive biomarkers for communicable and non-communicable diseases. This study aims to characterize and compare small RNA profiles in human biofluids. For this purpose, RNA was extracted from plasma and breast milk samples from 15 healthy postpartum mothers. Small RNA libraries were prepared with the NEBNext(R) small RNA library preparation kit and sequenced in an Illumina HiSeq2000 platform. miRNAs, isomiRs and clusters of small RNAs were annotated using seqBuster/seqCluster framework in 5 plasma and 10 milk samples that passed the initial quality control. The RNA yield was 81 ng/mL [standard deviation (SD): 41] and 3985 ng/mL (SD: 3767) for plasma and breast milk, respectively. Mean number of good quality reads was 4.04 million (M) (40.01% of the reads) in plasma and 12.5M (89.6%) in breast milk. One thousand one hundred eighty two miRNAs, 12,084 isomiRs and 1,053 small RNA clusters that included piwi-interfering RNAs (piRNAs), tRNAs, small nucleolar RNAs (snoRNA) and small nuclear RNAs (snRNAs) were detected. Samples grouped by biofluid, with 308 miRNAs, 1,790 isomiRs and 778 small RNA clusters differentially detected. In summary, plasma and milk showed a different small RNA profile. In both, miRNAs, piRNAs, tRNAs, snRNAs, and snoRNAs were identified, confirming the presence of non-miRNA species in plasma, and describing them for the first time in milk.

Hormone-refractory prostate cancer frequently relapses from therapy and inevitably progresses to a bone-metastatic status with no cure. Understanding of the molecular mechanisms conferring resistance to androgen deprivation therapy has the potential to lead to the discovery of novel therapeutic targets for type of prostate cancer with poor prognosis. Progression to castration-resistant prostate cancer (CRPC) is characterized by aberrant androgen receptor (AR) expression and persistent AR signaling activity. Alterations in metabolic activity regulated by oncogenic pathways, such as c-Myc, were found to promote prostate cancer growth during the development of CRPC. Non-coding RNAs represent a diverse family of regulatory transcripts that drive tumorigenesis of prostate cancer and various other cancers by their hyperactivity or diminished function. A number of studies have examined differentially expressed non-coding RNAs in each stage of prostate cancer. Herein, we highlight the emerging impacts of microRNAs and long non-coding RNAs linked to reactivation of the AR signaling axis and reprogramming of the cellular metabolism in prostate cancer. The translational implications of non-coding RNA research for developing new biomarkers and therapeutic strategies for CRPC are also discussed.

MicroRNAs usually regulate gene expression negatively, and aberrant expression has been involved in the development of several types of cancers. Microarray profiling of microRNA expression was performed to define a microRNA signature in a series of mycosis fungoides tumor stage (MFt, n=21) and CD30+ primary cutaneous anaplastic large cell lymphoma (CD30+ cALCL, n=11) samples in comparison with inflammatory dermatoses (ID, n=5). Supervised clustering confirmed a distinctive microRNA profile for cutaneous T-cell lymphoma (CTCL) with respect to ID. A 40 microRNA signature was found in MFt including upregulated onco-microRNAs (miR-146a, miR-142-3p/5p, miR-21, miR-181a/b, and miR-155) and downregulated tumor-suppressor microRNAs (miR-200ab/429 cluster, miR-10b, miR-193b, miR-141/200c, and miR-23b/27b). Regarding CD30+ cALCL, 39 differentially expressed microRNAs were identified. Particularly, overexpression of miR-155, miR-21, or miR-142-3p/5p and downregulation of the miR-141/200c clusters were observed. DNA methylation in microRNA gene promoters, as expression regulatory mechanism for deregulated microRNAs, was analyzed using Infinium 450K array and approximately one-third of the differentially expressed microRNAs showed significant DNA methylation differences. Two different microRNA methylation signatures for MFt and CD30+ cALCL were found. Correlation analysis showed an inverse relationship for microRNA promoter methylation and microRNA expression. These results reveal a subgroup-specific epigenetically regulated microRNA signatures for MFt and CD30+ cALCL patients.