Identification of differential and functionally active miRNAs in both anaplastic lymphoma kinase (ALK)+ and ALK? anaplastic large-cell lymphoma

Aberrant anaplastic lymphoma kinase (ALK) expression is a defining feature of many human cancers and was identified first in anaplastic large-cell lymphoma (ALCL), an aggressive non-Hodgkin T-cell lymphoma. Since that time, many studies have set out to identify the mechanisms used by aberrant ALK toward tumorigenesis. We have identified a distinct profile of micro-RNAs (miRNAs) that characterize ALCL; furthermore, this profile distinguishes ALK⁺ from ALK⁻ subtypes, and thus points toward potential mechanisms of tumorigenesis induced by aberrant ALK. Using a nucleophosmin-ALK transgenic mouse model as well as human primary ALCL tumor tissues and human ALCL-derived cell lines, we reveal a set of overlapping deregulated miRNAs that might be implicated in the development and progression of ALCL. Importantly, ALK⁺ and ALK⁻ ALCL could be distinguished by a distinct profile of “oncomirs”: Five members of the miR-17–92 cluster were expressed more highly in ALK⁺ ALCL, whereas miR-155 was expressed more than 10-fold higher in ALK⁻ ALCL. Moreover, miR-101 was down-regulated in all ALCL model systems, but its forced expression attenuated cell proliferation only in ALK⁺ and not in ALK⁻ cell lines, perhaps suggesting different modes of ALK-dependent regulation of its target proteins. Furthermore, inhibition of mTOR, which is targeted by miR-101, led to reduced tumor growth in engrafted ALCL mouse models. In addition to future therapeutical and diagnostic applications, it will be of interest to study the physiological implications and prognostic value of the identified miRNA profiles.     

Evolución de la investigación cardiológica española y análisis comparativo mundial de temas de especial relevancia

Introduction and objectivesWe used bibliometric techniques to analyze the participation of Spanish institutions in research on major cardiovascular topics during the last 4 decades.MethodsBibliometric indicators of production, collaboration and impact were obtained from the Science Citation Index Expanded (SCIE) database. Search strategies were used in major topics and institutional collaboration networks were identified, represented using the Kamada-Kawai algorithm.ResultsGlobal cardiovascular publications doubled from 2000 to 2018. In 2018, those by Spanish authors represented 2.33%, with a participation of between 7% and 1.84%, depending on the topics analyzed. The offset with respect to global production was between 0 and 7 years. Annual growth rates were higher in more recent topics. Revista Española de Cardiología published the largest number of articles from Spanish institutions. The journals generating the highest number of citations in the chosen topics were the Journal of the American College of Cardiology, Europace, and the European Heart Journal. Analysis of collaboration revealed a close interrelation between Spanish and foreign institutions, as well as groups with high production publishing independently.ConclusionsThe analysis disaggregated by subject showed the sustained growth of Spanish cardiovascular scientific production and more rapid growth in recently appearing topics. Collaboration networks showed a high degree of interrelation between Spanish and foreign institutions, including hospitals, universities, research institutes, and scientific societies.     

Comparing Efficiency of micro-RNA and mRNA Biomarker Liberation with Microbubble-Enhanced Ultrasound Exposure

Blood biomarkers are potentially powerful diagnostic tools that are limited clinically by low concentrations, the inability to determine biomarker origin and unknown patient baseline. Recently, ultrasound has been shown to liberate proteins and large mRNA biomarkers, overcoming many of these limitations. We have since demonstrated that adding lipid-stabilized microbubbles elevates mRNA concentration an order of magnitude compared with ultrasound without microbubbles, in vitro. Unfortunately the large size of some mRNA molecules may limit efficiency of release and hinder efficacy as an ultrasound-liberated biomarker. We hypothesize that smaller molecules will be released more efficiently with ultrasound than larger molecules. Although investigation of large libraries of biomarkers should be performed to fully validate this hypothesis, we focus on a small subset of mRNA and micro-RNAs. Specifically, we focus on miR-21 (22 base pairs [bp]), which is upregulated in certain forms of cancer, compared with previously investigated mammaglobin mRNA (502 bp). We also report release of micro-RNA miR-155 (22 bp) and housekeeping rRNA S18 (1869 bp). More than 10 million additional miR-21 copies per 100,000 cells are released with ultrasound-microbubble exposure. The low- molecular-weight miR-21 proved to be liberated 50 times more efficiently than high-molecular-weight mammaglobin mRNA, releasing orders of magnitude more miR-21 than mammaglobin mRNA under comparable conditions.     

Loss of miR-183/96 Alters Synaptic Strength via Presynaptic and Postsynaptic Mechanisms at a Central Synapse

A point mutation in miR-96 causes non-syndromic progressive peripheral hearing loss and alters structure and physiology of the central auditory system. To gain further insight into the functions of microRNAs (miRNAs) within the central auditory system, we investigated constitutive Mir-183/96^dko mice of both sexes. In this mouse model, the genomically clustered miR-183 and miR-96 are constitutively deleted. It shows significantly and specifically reduced volumes of auditory hindbrain nuclei, because of decreases in cell number and soma size. Electrophysiological analysis of the calyx of Held synapse in the medial nucleus of the trapezoid body (MNTB) demonstrated strongly altered synaptic transmission in young-adult mice. We observed an increase in quantal content and readily releasable vesicle pool size in the presynapse while the overall morphology of the calyx was unchanged. Detailed analysis of the active zones (AZs) revealed differences in its molecular composition and synaptic vesicle (SV) distribution. Postsynaptically, altered clustering and increased synaptic abundancy of the AMPA receptor subunit GluA1 was observed resulting in an increase in quantal amplitude. Together, these presynaptic and postsynaptic alterations led to a 2-fold increase of the evoked excitatory postsynaptic currents in MNTB neurons. None of these changes were observed in deaf Cldn14^ko mice, confirming an on-site role of miR-183 and miR-96 in the auditory hindbrain. Our data suggest that the Mir-183/96 cluster plays a key role for proper synaptic transmission at the calyx of Held and for the development of the auditory hindbrain.SIGNIFICANCE STATEMENT The calyx of Held is the outstanding model system to study basic synaptic physiology. Yet, genetic factors driving its morphologic and functional maturation are largely unknown. Here, we identify the Mir-183/96 cluster as an important factor to regulate its synaptic strength. Presynaptically, Mir-183/96^dko calyces show an increase in release-ready synaptic vesicles (SVs), quantal content and abundance of the proteins Bassoon and Piccolo. Postsynaptically, the quantal size as well as number and size of GluA1 puncta were increased. The two microRNAs (miRNAs) are thus attractive candidates for regulation of synaptic maturation and long-term adaptations to sound levels. Moreover, the different phenotypic outcomes of different types of mutations in the Mir-183 cluster corroborate the requirement of mutation-tailored therapies in patients with hearing loss.     

分化程度不同的鼻咽癌细胞株X线辐射后miR-7的表达差异

目的 探讨不同分化程度的鼻咽癌细胞株X线辐射后miR-7表达的变化.方法 2种细胞均分为对照组(未辐射)、2 Gy组、6 Gy组和10 Gy组,X线辐射细胞株,线性二次模型拟合CNE-1、CNE-2细胞存活曲线,Trizol法提取细胞总RNA.茎环法特异性逆转录miR-7,染料法实时定量PCR,以对照组CNE-1细胞为参照样本,△△Ct法得出各样品miR-7的相对倍数.结果 对照组CNE-2细胞miR-7的量是CNE-1的2.51倍.2 Gy组CNE-1细胞miK-7表达量增加,CNE-2细胞则由早期的49.44下降到晚期的0.15.6 Gy组两种细胞miR-7含量差别不大.10 Gy组CNE-1的miR-7含量高于CNE-2. 结论 分化程度不同的鼻咽癌细胞经不同剂量的X线辐射后,miR-7表达量各不相同.分化程度不同的鼻咽癌细胞的放射敏感性差异,可能是由调控miR-7表达能力不同造成的.     

Aspectos inmunogenéticos de la psoriasis con énfasis en micro-ARN

Psoriasis is a chronic inflammatory disease of the skin, of autoimmune origin, with different cells implicated in the aetiopathology, such as T helper lymphocytes (Th1 and Th17), keratinocytes, and cytokines produced by these cells. The epigenetic regulatory mechanisms are the junction between environmental exposure and genetic factors. It is known that microRNAs (miRNAs), single chain RNAs, are actively involved in epigenetic regulation. Alterations in the miR-125b, miR-424, miR- 21 and miR-203 expression, and others, have been involved in different aspects of the disease. Global studies of miRNA expression performed using microarrays and by direct RNA sequencing revealed important differences in miRNA expression in normal skin and psoriatic individuals. These miRNAs can be considered as potential therapeutic targets or biomarkers of disease.     

胃癌组织中miR-181a及其靶基因KLF6表达变化及意义

目的观察胃癌组织中微小RNA(miRNA)-181a(miR-181a)与其靶基因Kruppel样因子6(Kruppel like factor 6,KLF6,又称锌指转录因子9或核心启动子元件结合蛋白)的表达变化,并探讨其意义。方法 9例胃癌患者,手术时留取癌组织及癌旁组织,抽提其中的总RNA及蛋白质,采用实时荧光定量PCR检测标本中的miR-181a,Western blot检测KLF6蛋白。结果胃癌组织中miR-181a的表达量(2-△△Ct)为(1.981±1.080),癌旁组织miR-181a的表达量为(0.394±0.093);癌组织KLF6蛋白灰度值为(0.241 6±0.135 5),癌旁组织为(0.540 2±0.05 5),两种组织中的miR-181a、KLF6蛋白表达量相比,均p﹤0.01;miR-181a与KLF6蛋白的表达呈负相关(r=-0.590,p﹤0.01)。结论胃癌组织中miR-181a表达明显上调,KLF6蛋白表达明显下降;结合前期实验研究,我们推测miR-181a可能通过转录后基因沉默机制使KLF6蛋白表达下降,从而促进胃癌的发生发展。