Exosomes as Tools to Suppress Primary Brain Tumor

Exosomes are small microvesicles released by cells that efficiently transfer their molecular cargo to other cells, including tumor. Exosomes may pass the blood–brain barrier and have been demonstrated to deliver RNAs contained within to brain. As they are non-viable, the risk profile of exosomes is thought to be less than that of cellular therapies. Exosomes can be manufactured at scale in culture, and exosomes can be engineered to incorporate therapeutic miRNAs, siRNAs, or chemotherapeutic molecules. As natural biological delivery vehicles, interest in the use of exosomes as therapeutic delivery agents is growing. We previously demonstrated a novel treatment whereby mesenchymal stromal cells were employed to package tumor-suppressing miR-146b into exosomes, which were then used to reduce malignant glioma growth in rat. The use of exosomes to raise the immune system against tumor is also drawing interest. Exosomes from dendritic cells which are antigen-presenting, and have been used for treatment of brain tumor may be divided into three categories: (1) exosomes for immunomodulation-based therapy, (2) exosomes as delivery vehicles for anti-tumor nucleotides, and (3) exosomes as drug delivery vehicles. Here, we will provide an overview of these three applications of exosomes to treat brain tumor, and examine their prospects on the long road to clinical use.     

miRNAs as biomarkers of atrial fibrillation

Abstract

Atrial fibrillation (AF) is a highly prevalent arrhythmia with pronounced morbidity and mortality. Genetics analysis has established electrophysiological substrates, which determine individual vulnerability to AF occurrence and maintenance. MicroRNAs (miRNAs) found in virtually all organisms function as negative regulators of protein-coding genes. Several studies have suggested a role for miRNAs in the regulation of cardiac excitability and arrhythmogenesis. This review is based on 18 studies conducted between 2009 and 2013 to investigate the association of miRNAs with AF. miRNAs are discussed here as candidate biomarkers for AF in blood and cardiac tissues and as potential targets for AF therapy.     

Atrial remodeling in atrial fibrillation and association between microRNA network and atrial fibrillation

Atrial fibrillation (AF) remains one of the leading causes of morbidity and mortality in the world which are related to palpitations, fainting, congestive heart failure or stroke. The mechanism for atrial fibrillation has been identified as electrical remodeling, structure remodeling and intracellular calcium handling remodeling. microRNAs (miRNAs) have recently emerged as one of the important factors in regulating gene expression. So far, thousands of miRNA genes have been found in diverse animals with the function of regulating cell death, cell proliferation, haematopoiesis and even participate in the processing of cardiovascular disease. In this review, we summarize the mechanism of AF and the association of microRNAs network with AF. We provide a potential perspective of miRNAs as the therapeutic target for AF.     

Green tea may be benefit to the therapy of atrial fibrillation

Atrial fibrillation (AF) is the most common cardiac arrhythmia in clinical practice. Systemic inflammatory state, oxidative stress injury, and atrial fibrosis are identified as the main mechanisms for AF. Considering the multifactorial mechanisms of AF, a novel therapeutic agent with multi-bioactivities should be presented. Regular consumption of green tea has been associated with a reduced risk of coronary heart disease and against a large number of pathologic conditions. Recent results indicate that green tea extract, especially (-)-epigallocatechin-3-gallate, could effectively decrease inflammatory factors secretion, antagonize oxidation, and inhibit matrix metalloproteinase activities. Inhibition of inflammation, modulation of oxidative stress, and targeting tissue fibrosis represent new approaches in tackling AF; therefore, green tea may be an innovative therapeutic candidate to prevent the occurrence, maintenance, and recurrence of AF.     

Exosomal microRNA and stroke: A review

Blood vessels rupture or occlusion in brain results in stroke. Stroke is the major reason for mortality and dysfunction worldwide. Despite several attempts, there are no any approved therapeutic approaches for stroke subjects. The most neuroprotective agents showed the positive effects in preclinical reports, while there are no significant therapeutic impacts in the clinical trials. MicroRNAs (miRNAs) are small noncoding RNAs which involved in the modulation of a variety of cellular and molecular pathways. Given that deregulation of these molecules is related to initiation and progression of stroke. Exosomes are nano-carriers which are able to transfer different cargos such as miRNAs to recipient cells. Increasing evidence revealed that exosomal miRNAs are one of very important factors which are involved in the pathogenesis of stroke. Hence, more understanding about the role of exosomal miRNAs in stroke pathogenesis could contribute in discovering and developing new therapeutic approaches. Moreover, it has been proved the exosomal miRNAs could be used as noninvasive biomarkers in diagnosis and monitoring response to therapy in subjects with stroke. Herein for first time, we summarized different exosomal miRNAs involved in pathogenesis of stroke.     

MicroRNA replacement therapy in cancer

Despite the recent progress in cancer management approaches, the mortality rate of cancer is still growing and there are lots of challenges in the clinics in terms of novel therapeutics. MicroRNAs (miRNA) are regulatory small noncoding RNAs and are already confirmed to have a great role in regulating gene expression level by targeting multiple molecules that affect cell physiology and disease development. Recently, miRNAs have been introduced as promising therapeutic targets for cancer treatment. Regulatory potential of tumor suppressor miRNAs, which enables regulation of entire signaling networks within the cells, makes them an interesting option for developing cancer therapeutics. In this regard, over recent decades, scientists have aimed at developing powerful and safe targeting approaches to restore these suppressive miRNAs in cancerous cells. The present review summarizes the function of miRNAs in tumor development and presents recent findings on how miRNAs have served as therapeutic agents against cancer, with a special focus on tumor suppressor miRNAs (mimics). Moreover, the latest investigations on the therapeutic strategies of miRNA delivery have been presented.     

Anti-M2 muscarinic acetylcholine receptor autoantibodies: New therapeutic targets in atrial fibrillation

Atrial fibrillation is a major and growing public health problem in the world. Heart failure is one of the most common clinical settings for chronic AF. The mechanism underlying AF is still not fully understood. Increasing evidence suggests that activation of G-protein-coupled receptors by autoantibodies may contribute to the pathophysiology of cardiac diseases and stimulatory anti-M2 muscarinic acetylcholine receptor autoantibodies (M2-AABs) were detected in several heart diseases such as dilated cardiomyopathy and Chagas's heart disease. Functional studies have demonstrated that these autoantibodies are able not only to bind to target receptors in the myocardium, but also to induce receptor-mediated biological responses, as partial agonists, which triggers neurotransmitter-receptor-mediated biological effects. Clinical investigation shows that AF was more common in M2-AAB-positive than in M2-AAB-negative patients. Thus, it is a reasonable hypothesis that M2-AABs may participate in the induction and perpetuation of AF. Studies to confirm this hypothesis may lead to new directions in the study of the pathogenesis of AF and the development of a new therapeutic option for this disorder.     

Exosomal microRNAs from mesenchymal stem/stromal cells: Biology and applications in neuroprotection

Mesenchymal stem/stromal cells (MSCs) are extensively studied as cell-therapy agents for neurological diseases. Recent studies consider exosomes secreted by MSCs as important mediators for MSCs’ neuroprotective functions. Exosomes transfer functional molecules including proteins, lipids, metabolites, DNAs, and coding and non-coding RNAs from MSCs to their target cells. Emerging evidence shows that exosomal microRNAs (miRNAs) play a key role in the neuroprotective properties of these exosomes by targeting several genes and regulating various biological processes. Multiple exosomal miRNAs have been identified to have neuroprotective effects by promoting neurogenesis, neurite remodeling and survival, and neuroplasticity. Thus, exosomal miRNAs have significant therapeutic potential for neurological disorders such as stroke, traumatic brain injury, and neuroinflammatory or neurodegenerative diseases and disorders. This review discusses the neuroprotective effects of selected miRNAs (miR-21, miR-17-92, miR-133, miR-138, miR-124, miR-30, miR146a, and miR-29b) and explores their mechanisms of action and applications for the treatment of various neurological disease and disorders. It also provides an overview of state-of-the-art bioengineering approaches for isolating exosomes, optimizing their yield and manipulating the miRNA content of their cargo to improve their therapeutic potential.     

Exploiting the therapeutic potential of microRNAs in viral diseases: expectations and limitations

New therapeutic approaches are urgently needed for serious diseases, including cancer, cardiovascular diseases, viral infections, and others. A recent direction in drug development is the utilization of nucleic acid-based therapeutic molecules, such as antisense oligonucleotides, ribozymes, short interfering RNA (siRNA), and microRNA (miRNA). miRNAs are endogenous, short, non-coding RNA molecules. Some viruses encode their own miRNAs, which play pivotal roles in viral replication and immune evasion strategies. Conversely, viruses that do not encode miRNAs may manipulate host cell miRNAs for the benefits of their replication. miRNAs have therefore become attractive tools for the study of viral pathogenesis. Lately, novel therapeutic strategies based on miRNA technology for the treatment of viral diseases have been progressing rapidly. Although this new generation of molecular therapy is promising, there are still several challenges to face, such as targeting delivery to specific tissues, avoiding off-target effects of miRNAs, reducing the toxicity of the drugs, and overcoming mutations and drug resistance. In this article, we review the current knowledge of the role and therapeutic potential of miRNAs in viral diseases, and discuss the limitations of these therapies, as well as strategies to overcome them to provide safe and effective clinical applications of these new therapeutics.     

外泌体介导的靶向基因运送及在心血管疾病中的应用

外泌体是体内几乎所有细胞分泌的具有双层脂质膜结构的纳米级小囊泡。外泌体大小均匀,平均直径为40～120 nm,存在于所有体液中。外泌体曾一度被认为是细胞成熟过程中清除废弃细胞器的‘垃圾袋’。但近年研究显示：外泌体含有丰富的来源于‘供体细胞’的信号分子,如蛋白质、DNA、mRNA、miRNA以及lncRNA等。当外泌体与‘受体细胞’融合时,这些信号分子便被运送到‘受体细胞’,从而实现细胞 细胞之间的通讯,影响‘受体细胞’的生理病理过程。虽然外泌体的研究目前主要集中在癌症等疾病的预防、诊断与治疗中,但是越来越多的研究显示,外泌体在心血管系统的生理及病理过程中同样发挥着重要作用。本文将对外泌体的起源、分离与纯化方法及外泌体介导的‘细胞 细胞’之间的通讯机制进行综述,并重点论述利用基因工程技术对外泌体进行靶向运输的方法及靶向外泌体运送在心血管疾病治疗中的应用。     

Expression of exosomal microRNAs during chondrogenic differentiation of human bone mesenchymal stem cells

The aim of the current study was to compare the expression of microRNAs (miRNAs) in exosomes derived from human bone mesenchymal stem cells (hBMSCs) with and without chondrogenic induction. Exosomes derived from hBMSCs were isolated and identified. Microarray analysis was performed to compare miRNA expression between exosomes derived from hBMSCs with and without chondrogenic induction, and quantitative real-time polymerase chain reaction (qRT-PCR) was used to verify the differentially expressed miRNAs. hBMSCs were transfected with miRNA mimic to extract miRNA-overexpressed exosomes. The results showed that most exosomes exhibited a cup-shaped or round-shaped morphology with a diameter of approximately 50-200 nm and expressed CD9 and CD63. We detected 141 miRNAs that were differentially expressed with and without chondrogenic induction by over a twofold change, including 35 upregulated miRNAs, such as miR-1246, miR-1290, miR-193a-5p, miR-320c, and miR-92a, and 106 downregulated miRNAs, such as miR-377-3p and miR-6891-5p. qRT-PCR analysis validated these results. Exosomes derived from hBMSCs overexpressing miR-320c were more efficient than normal exosomes derived from control hBMSCs at promoting osteoarthritis chondrocyte proliferation, down-regulated matrix metallopeptidase 13 and up-regulated (sex determining region Y)-box 9 expression during hBMSC chondrogenic differentiation. In conclusion, we identified a group of upregulated miRNAs in exosomes derived from hBMSCs with chondrogenic induction that may play an important role in mesenchymal stem cell-derived exosomes in cartilage regeneration and, ultimately, the treatment of arthritis. We demonstrated the potential of these modified exosomes in the development of novel therapeutic strategies.     

Beat shock proteins and atrial fibrillation

In this mini-review, the role of heat shock proteins in susceptability to induction of atrial fibrillation (AF) or in the process of AF is discussed. AF is the most common arrhythmia in humans, is self-perpetuating in nature and hence tends to become more persistent in time. Some studies show a correlation between high Hsp70 (HspA1A) expression in cardiac tissue and a reduced susceptability to induction of postoperative AF. Expression of Hsp70, Hsc70 (HspA8), Hsp40 (DnaJB1), Hsp60 (HspD1), Hsp90 (HspC1) was not associated with progression of AF. However, both correlative studies in human and experimental studies suggest that Hsp27 (HspB1) may delay progression of AF to the more permanent forms and hence Hsp27 might be referred to as a "Beat shock protein".     

Neuronal Differentiation of Human Mesenchymal Stem Cells Using Exosomes Derived from Differentiating Neuronal Cells

Exosomes deliver functional proteins and genetic materials to neighboring cells, and have potential applications for tissue regeneration. One possible mechanism of exosome-promoted tissue regeneration is through the delivery of microRNA (miRNA). In this study, we hypothesized that exosomes derived from neuronal progenitor cells contain miRNAs that promote neuronal differentiation. We treated mesenchymal stem cells (MSCs) daily with exosomes derived from PC12 cells, a neuronal cell line, for 1 week. After the treatment with PC12-derived exosomes, MSCs developed neuron-like morphology, and gene and protein expressions of neuronal markers were upregulated. Microarray analysis showed that the expression of miR-125b, which is known to play a role in neuronal differentiation of stem cells, was much higher in PC12-derived exosomes than in exosomes from B16-F10 melanoma cells. These results suggest that the delivery of miRNAs contained in PC12-derived exosomes is a possible mechanism explaining the neuronal differentiation of MSC.     

Efficient and specific inhibition of plant microRNA function by anti-microRNA oligonucleotides (AMOs) in vitro and in vivo

Key message

Anti-microRNA oligonucleotides (AMOs) are efficient and sequence-specific inhibitors of plant miRNA function both in vitro and in vivo.

Abstract

MicroRNAs (miRNAs) are small non-coding RNAs that play critical roles in developmental and physiological processes in plants and animals. Although miRNA knockdown by chemically modified antisense oligonucleotides prevails in animal and therapeutic studies, no such application has ever been reported in plants. Here, we show that sucrose-mediated delivery of 2′-O-methyl (2′-O-Me) anti-miRNA oligonucleotides (AMOs) is an efficient and sequence-specific way of inhibiting plant miRNA activity both in vitro and in vivo. Administration of AMOs to rice protoplasts and intact leaves resulted in efficient inhibition of miRNAs with concurrent de-repression of their target genes. AMOs caused simultaneous inhibition of miRNAs from the same family but exerted negligible effects on miRNAs from different families. In rice seedlings, a single-dose AMO treatment conferred long-lasting miRNA inhibition for at least 7 days. Although simultaneous dysregulation of multiple miRNAs by an AMO-and-miRNA-mimic mixture resulted in severe root defects, the phenotypic effects of individual AMOs and miRNA mimics were negligible, suggesting that those miRNAs function together in regulatory networks to ensure homeostasis. Our results validate the utility of AMOs as an efficient tool for plant miRNA loss-of-function studies in vivo, and this approach may prove to be a highly promising general method for unraveling miRNA-mediated gene-regulatory networks.

     

Viral delivery of miR-196a ameliorates the SBMA phenotype via the silencing of CELF2

Spinal and bulbar muscular atrophy (SBMA) is an inherited neurodegenerative disorder caused by the expansion of the polyglutamine (polyQ) tract of the androgen receptor (AR-polyQ). Characteristics of SBMA include proximal muscular atrophy, weakness, contraction fasciculation and bulbar involvement. MicroRNAs (miRNAs) are a diverse class of highly conserved small RNA molecules that function as crucial regulators of gene expression in animals and plants. Recent functional studies have shown the potent activity of specific miRNAs as disease modifiers both in vitro and in vivo. Thus, potential therapeutic approaches that target the miRNA processing pathway have recently attracted attention. Here we describe a novel therapeutic approach using the adeno-associated virus (AAV) vector–mediated delivery of a specific miRNA for SBMA. We found that miR-196a enhanced the decay of the AR mRNA by silencing CUGBP, Elav-like family member 2 (CELF2). CELF2 directly acted on AR mRNA and enhanced the stability of AR mRNA. Furthermore, we found that the early intervention of miR-196a delivered by an AAV vector ameliorated the SBMA phenotypes in a mouse model. Our results establish the proof of principle that disease-specific miRNA delivery could be useful in neurodegenerative diseases.     

Modern conceptions of the atrial fibrillation development. The role of the myocardial sleeves in the pulmonary veins

Atrial fibrillation (AF) is the most common supraventricular cardiac arrhythmia. In this review several conceptions focused on the mechanisms of the AF initiation are discussed. At present time viewpoint that the ectopical activity in the pulmonary vien myocardial sleeves (PVs) account for AF in prevailing. PVs myocardium has been the subject of many anatomical and physiological investigations. PVs myocardium differs from left atria tissue and has many moprhological properties that make in convenient substrate for AF initiation and maintenance. PVs cardiomyocytes were shown to have great variability of electrophysiological properties (action potential duration, resting potential, upstroke velocity, etc.). Attempt to discuss afterdepolarization, triggered activity and abnormal automaticity as initiators of AF in PVs was made. It was shown that as in experimental condition, as in vivo in PVs can exist er-entry. Possibly, re-entry from PVs could be the one mechanism by which AF is initiated. In review big attention to the innervations of PVs and role of the sympathetic and the parasympathetic nerves in PVs ectopical activity is paid. Combined influence of autonomic nerves may be critical to initiating AF in PVs. Pharmacological intervention as a possible way to suppress or prevent the activity in the PVs that leads to AF is discussed.