MicroRNAs and exosomes: promising new biomarkers in acute myeloid leukemias?

ABSTRACT Despite advances in understanding of carcinogenesis and of treatment of acute myeloid leukemia, this neoplasm still has a lethality of at least 30%. The search for biomarkers that can predict the response to treatment in the early stages of the disease is still necessary. In recent years, a new form of cellular communication between tumor and non-neoplastic cells has been discovered: the exchange of information through extracellular vesicles. These are small vesicles released by membrane-coated cells that carry proteins, lipids, messenger RNAs, microRNA and DNA, which can be internalized and promote biological changes in target cells. Exosomes are qualified as a type of extracellular vesicle and, in tumors, carry immunoinhibitory signals that promote the escape of immune control. Recent studies have showed their involvement in communication with the cells of the tumor microenvironment and with chemoresistance in several tumors. To date, there is no information about immunoregulatory microRNAs transported by exosomes and their correlation with clinical evolution during chemotherapy for acute myeloid leukemia. Knowledge about immunomodulatory microRNAs obtained by leukemic cells and transported by exosomes can direct us towards the design of new diagnostic and treatment tools in this type of leukemia.

Research progress of extracellular vesicles in the treatment of renal disease / 生理学报

Extracellular vesicles (EVs) are lipid bilayer-enclosed structures containing diverse bioactive cargoes that play a major role in intercellular communication in both physiological and pathological conditions. Currently, the field of EV-based therapy has been rapidly growing, and two main therapeutic uses of EVs can be surmised: (i) exploiting stem cell-derived EVs as therapeutic agents; and (ii) employing EVs as natural therapeutic vectors for drug delivery. This review will discuss the recent advances in EV-based therapy in the treatment of renal disease.

Exosomal microRNAs: an emerging player in systemic lupus erythematosus / 生理学报

Exosomes are nanometer-sized membranous extracellular vesicles that can be secreted by almost all types of cells in the body. Exosomes are involved in cell-to-cell communication through autocrine and paracrine forms. Exosomal microRNAs (miRNAs) are stable in plasma, urine and other body fluids, and have various biological functions. They play an irreplaceable role in the occurrence, development, immune regulation of systemic lupus erythematosus (SLE). Recent studies have proposed that exosomal miRNAs have promising application prospects in the pathogenesis, early diagnosis, and treatment of SLE. Therefore, this review aims to introduce the current research progress on exosomal miRNAs in SLE and analyze their potential application value.

Transforming growth factor-β1-induced N-cadherin drives cell-cell communication through connexin43 in osteoblast lineage / 国际口腔科学杂志·英文版

Gap junction (GJ) has been indicated to have an intimate correlation with adhesion junction. However, the direct interaction between them partially remains elusive. In the current study, we aimed to elucidate the role of N-cadherin, one of the core components in adhesion junction, in mediating connexin 43, one of the functional constituents in gap junction, via transforming growth factor-β1(TGF-β1) induction in osteoblasts. We first elucidated the expressions of N-cadherin induced by TGF-β1 and also confirmed the upregulation of Cx43, and the enhancement of functional gap junctional intercellular communication (GJIC) triggered by TGF-β1 in both primary osteoblasts and MC3T3 cell line. Colocalization analysis and Co-IP experimentation showed that N-cadherin interacts with Cx43 at the site of cell-cell contact. Knockdown of N-cadherin by siRNA interference decreased the Cx43 expression and abolished the promoting effect of TGF-β1 on Cx43. Functional GJICs in living primary osteoblasts and MC3T3 cell line were also reduced. TGF-β1-induced increase in N-cadherin and Cx43 was via Smad3 activation, whereas knockdown of Smad3 signaling by using siRNA decreased the expressions of both N-cadherin and Cx43. Overall, these data indicate the direct interactions between N-cadherin and Cx43, and reveal the intervention of adhesion junction in functional gap junction in living osteoblasts.

Effect of exosome / 中南大学学报(医学版)

Traumatic brain injury (TBI) is a main cause of death and disability worldwide, posing a serious threat to public health. But currently, the diagnosis and treatments for TBI are still very limited. Exosomes are a group of extracellular vesicles and participate in multiple physiological processes including intercellular communication and substance transport. Non-coding RNAs (ncRNA) are of great abundancy as cargo of exosomes. Previous studies have shown that ncRNAs are involved in several pathophysiological processes of TBI. However, the concrete mechanisms involved in the effects induced by exosome-derived ncRNA remain largely unknown. As an important component of exosomes, ncRNA is of great significance for diagnosis, precise treatment, response evaluation, prognosis prediction, and complication management after TBI.

Effect of the Connexin 43 Coupling to the Biobehavior of Multiple Myeloma Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of gap junction intercellular communication (GJIC) combined by connexin43 (Cx43) and its signal to the biobehavior of multiple myeloma (MM) cells, and its possible mechanism.@*METHODS@#The mesenchymal stem cell (MSC) cells were isolated and cultured from patients with MM and normal donors. The expression of connexin43 (Cx43) in MSC cells from different sources was detected by RT-PCR and Western blot. The side population (SP) cells were sorted by flow cytometry (FCM). The effect of MSC cells from different sources to the cell cycle, Cx43 expression, colony formation in vitro, stem cell related genes expression, cytokines secretion and chemoresistance in MM SP cells as well as with or without Cx43 inhibitor 18α-glycyrrhetinic acid (18α-GA) was observed.@*RESULTS@#There was no significantly difference between the MSC isolated from normal donor and MM patients. Western blot showed that Cx43 expression in SP cells was up-regulated when the cells were incubated with MSC, and medium containing 18α-GA could partially inhibit it, moreover, it was more significant in MSC cells of MM patients. The ability of colony formation of SP cells in vitro was higher than those of MM cells and MM-MSC could promote the colony formation in a co-culture manner. The effect of MM-MSC to SP cells was down-regulated after 18α-GA was added. RT-PCR showed that there was several important stem cell-related genes including c-myc, Oct-4 Klf-4, and Sox-2 were found in RPMI 8226 cells, but those cells were up-regulated in SP cells (P0.05). Cytometry bead array assays showed that MM-MSCs could secrete high level of IL-6, but the levels of IL-6, IL-10 and TGF-β increased significantly when the MM-MSCs were co-cultured with SP cells (P<0.05), especially the levels of IL-6 and IL-10 were significantly higher than cultured alone. There was no significant change in the levels of bFGF and IL-17 before and after co-cultured. The levels of IL-6, IL-10 and TGF-β in supernatant decreased significantly after GJ inhibitor 18α-GA was added. PI/Annexin V assay showed that MM cells were sensitive to bortezomib (BTZ)-induced apoptosis, but the sensitivity for SP cells was weaker. The ratio of cell apoptosis was 75.2%±0.77% and 8.12%±0.86% (P<0.001), respectively. MM-MSC could down-regulate the cell apoptosis induced by BTZ, while the sensitivity of MM cells to BTZ could be partially recovered after GJ inhibitor was added.@*CONCLUSION@#MSC derived from MM patients can enhance GJIC to maintain its "hematopoiesis" by up-regulating the expression of Cx43 in MM cells, and at the same time promote cell proliferation and drug recistance by secreting multiple cytokines, which finally contributes to the relapse of MM.

The luciferase reporter system of the MMP12 endogenous promoter for investigating transcriptional regulation of the human MMP12 gene

Background: Matrix metalloproteinase 12 (MMP12), a member of MMPs, can take lots of roles including extracellular matrix component degradation, viral infection, inflammation, tissue remodeling and tumorigenesis. To explore the transcriptional regulation of MMP12 gene, a sensitive luciferase reporter HEK293 cell line for endogenous MMP12 promoter was generated by CRISPR/Cas9 technology. Results: The HEK293-MMP12-T2A-luciferase-KI cell line was successfully established by CRISPR/Cas9 technology. The sequencing results indicated that one allele of the genome was proven to have a site-directed insertion of luciferase gene and another allele of the genome was confirmed to have additional 48 bp insertion in this cell line. The cell line was further demonstrated to be a sensitive reporter of the endogenous MMP12 promoter by applying transcription factors STAT3, AP-1 and SP-1 to the cell line. The reporter cell line was then screened with bioactive small molecule library, and a small molecule Tanshinone I was found to significantly inhibit the transcriptional activity of MMP12 gene in HEK293-MMP12-T2A-luciferase-KI cell line by luciferase activity assay, which was further confirmed to inhibit the expression of MMP12 mRNA in wild-type HEK293 cells. Conclusions: This novel luciferase knock-in reporter system will be helpful for investigating the transcriptional regulation of MMP12 gene and screening the drugs targeting MMP12 gene.

Perfil proteômico de bactérias presentes em canais radiculares de dentes com periodontite apical sintomática ou assintomática / Proteomic profile of bacteria present in root canals of teeth with symptomatic or asymptomatic apical periodontitis

As infecções endodônticas são causadas por uma comunidade multiespécie de bactérias. Com os métodos de identificação microbiológica e quantificação de endotoxinas, é difícil inferir a fisiologia, a função e a patogenicidade microbiana. Assim, a análise proteômica é uma técnica que pode revolucionar o estudo da patogênese das infecções endodônticas. O presente estudo tem por objetivo analisar o perfil proteômico de infecções endodônticas relacionadas a dentes com periodontite apical sintomática ou assintomática utilizando cromatografia líquida associada à espectrometria de Massas. A análise deste perfil proteômico visa a proporcionar a compreensão dos aspectos ecológicos e patogênicos do comportamento de comunidades bacterianas endodônticas através da identificação de proteínas expressas no referido ambiente no momento da coleta e a determinação da função dessas proteínas. Foram coletadas amostras de 18 pacientes encaminhados para tratamento de canal radicular ou tratamento de emergência na Faculdade de Odontologia de Araçatuba FOA ­ UNESP. Foram incluídos dentes com infecção endodôntica primária, sintomáticos ou assintomáticos. A identificação dos peptídeos foi feita num sistema nanoACQUITY UPLC-Xevo QTof MS system (Waters), a identificação das proteínas foi obtida utilizando o software Protein Lynx Global Server (PLGS) versão 3.0, utilizando o banco de dados de proteínas UniProtKB. A diferença de expressão entre os grupos foi obtida através do mesmo software, considerando-se p<0,05 para as proteínas subreguladas e 1-p>0,95 para as proteínas suprareguladas. Foram identificados um total de 2181 números de acessos entre fragmentos, isoformas e proteínas completas humana e 51 proteínas bacterianas e ambas foram classificadas quanto a sua função biológica, em relação às proteínas exclusivas de cada grupo, 347 proteínas foram identificadas no grupo sintomático. As funções biológicas mais prevalentes foram comunicação celular e transdução de sinais, seguidas pela resposta imune, observou-se diversas proteínas exclusivamente expressas no grupo sintomático, indicando a influência direta da periodontite sintomática na resposta do hospedeiro(AU)

Endodontic infections are caused by a multispecies community of bacteria. With microbiological identification methods and endotoxin quantification, it is difficult to infer physiology, function and microbial pathogenicity. Thus, proteomic analysis is a technique that can revolutionize the study of the pathogenesis of endodontic infections. The present study aims to analyze the proteomic profile of endodontic infections related to teeth with symptomatic or asymptomatic apical periodontitis using liquid chromatography associated with mass spectrometry. The analysis of this proteomic profile aims to provide an understanding of the ecological and pathogenic aspects of the behavior of endodontic bacterial communities by identifying proteins expressed in the said environment at the time of collection and determining the function of these proteins. Samples were collected from 18 patients referred for root canal treatment or emergency treatment at the School of Dentistry of Araçatuba FOA - UNESP. Teeth with primary endodontic infection, symptomatic or asymptomatic were included. The identification of peptides was made in a nanoACQUITY UPLC-Xevo QTof MS system (Waters) system, the identification of proteins was obtained using protein lynx global server (PLGS) software version 3.0, using the UniProtKB protein database. The difference in expression between the groups was obtained through the same software, considering p<0.05 for subregulated proteins and 1-p>0.95 for the superregulated proteins. A total of 2,181 access numbers were identified between human fragments, isoforms and complete proteins and 51 bacterial proteins, and both were classified as their biological function, in relation to the exclusive proteins of each group, 347 proteins were identified in the symptomatic group. The most prevalent biological functions were cellular communication and signal transduction, followed by the immune response, and several proteins were observed exclusively expressed in the symptomatic group, indicating the direct influence of symptomatic periodontitis on the host response(AU)

Gap junction-mediated cell-to-cell communication in oral development and oral diseases: a concise review of research progress / 国际口腔科学杂志·英文版

Homoeostasis depends on the close connection and intimate molecular exchange between extracellular, intracellular and intercellular networks. Intercellular communication is largely mediated by gap junctions (GJs), a type of specialized membrane contact composed of variable number of channels that enable direct communication between cells by allowing small molecules to pass directly into the cytoplasm of neighbouring cells. Although considerable evidence indicates that gap junctions contribute to the functions of many organs, such as the bone, intestine, kidney, heart, brain and nerve, less is known about their role in oral development and disease. In this review, the current progress in understanding the background of connexins and the functions of gap junctions in oral development and diseases is discussed. The homoeostasis of tooth and periodontal tissues, normal tooth and maxillofacial development, saliva secretion and the integrity of the oral mucosa depend on the proper function of gap junctions. Knowledge of this pattern of cell-cell communication is required for a better understanding of oral diseases. With the ever-increasing understanding of connexins in oral diseases, therapeutic strategies could be developed to target these membrane channels in various oral diseases and maxillofacial dysplasia.

Advances in study on exosomes and their applications in kidney diseases / 中南大学学报(医学版)

Exosomes are in a size of 30-100 nm vesicles released by various cells, with a double-layered lipid membrane containing DNA, RNA, and protein. In the past, exosomes were considered to be molecular waste, and recently exosomes have been shown to be involved in many pathophysiological processes, including intercellular communication, immune response, nerve repair, and tumorigenesis. Exosomes are present in numerous body fluids, and urinary exosomes have been shown to be biomarkers of a variety of kidney diseases.

New avenues for systematically inferring cell-cell communication: through single-cell transcriptomics data

For multicellular organisms, cell-cell communication is essential to numerous biological processes. Drawing upon the latest development of single-cell RNA-sequencing (scRNA-seq), high-resolution transcriptomic data have deepened our understanding of cellular phenotype heterogeneity and composition of complex tissues, which enables systematic cell-cell communication studies at a single-cell level. We first summarize a common workflow of cell-cell communication study using scRNA-seq data, which often includes data preparation, construction of communication networks, and result validation. Two common strategies taken to uncover cell-cell communications are reviewed, e.g., physically vicinal structure-based and ligand-receptor interaction-based one. To conclude, challenges and current applications of cell-cell communication studies at a single-cell resolution are discussed in details and future perspectives are proposed.

Role of neuron and non-neuronal cell communication in persistent orofacial pain

It is well known that trigeminal nerve injury causes hyperexcitability in trigeminal ganglion neurons, which become sensitized. Long after trigeminal nerve damage, trigeminal spinal subnucleus caudalis and upper cervical spinal cord (C1/C2) nociceptive neurons become hyperactive and are sensitized, resulting in persistent orofacial pain. Communication between neurons and non-neuronal cells is believed to be involved in these mechanisms. In this article, the authors highlight several lines of evidence that neuron-glial cell and neuron macrophage communication have essential roles in persistent orofacial pain mechanisms associated with trigeminal nerve injury and/or orofacial inflammation.

Research Advances on Role of Extracellular Vesicles in the Bone Marrow Microenvironment of Patients with Hematological Malignancies--Review / 中国实验血液学杂志

Bone marrow (BM) microenvironment appears to play an important role in the pathogenesis of hematological malignancies. Apart from soluble factors and direct cell-cell contact, the extracellular vesicles (EVs) were identified as a third mediator for cell communication within BM microenvironment. Recently, more and more evidences have demonstrated that EVs are also involved in the dysregulation of the BM microenvironment in patients with hematological malignancies. Therefore this review focuses on the biological characteristics of EVs, the clinical value of EVs as biomarkers, the BM microenvironment reprogramming in hematological malignancies by EVs, and the potential role of EVs in drug resistance and therapy of hematological malignancies.

The biological functions of cell-to-cell connection over long distance--membrane nanotube / 生理学报

Cell-to-cell connections provide conduits for signal exchanges, and play important functional roles in physiological and pathological processes of multicellular organisms. Membrane nanotubes are common long-distance connections between cells, not only transfer molecule signals and mitochondria, but also cooperate with gap junction and other cell-to-cell communications to transfer signals. During the last decade, there are many studies about membrane nanotubes, which focus on the similarities and differences between membrane nanotubes and other cell-to-cell communications, as well as their biological functions. In the present review, we summarized the latest findings about the structural diversity, the similarities and differences in signal transmission with other types of cell-to-cell communications, and physiological and pathological roles of membrane nanotubes.

The role of stem cell-derived exosomes in repairing myocardial injury / 生理学报

At present, it is generally believed that the paracrine effect of stem cells in the repair of myocardial injury is one of the important ways for stem cell therapy. Exosomes are phospholipid bilayer-enclosed nanovesicles that secreted by cells under physiological and pathological conditions. Cargo loaded into exosomes including protein, lipids and nucleic acids can be delivered to recipient cells. Therefore, exosomes are recognized as important mediators for intercellular communication. It has been suggested that exosomes from stem cells (eg. embryonic stem cells, induced pluripotent stem cells, cardiac progenitor cells, mesenchymal stem cells and cardiosphere-derived cells) have protective effects against heart injury. In this review, we summarized recent research progresses on stem cell-derived exosomes in myocardial injury, including the therapeutic effects and mechanism.

Exosomes and their roles in neurodegenerative diseases: from pathogenic, diagnostic and therapeutical perspectives / 生理学报

Exosomes are extracellular membranous vesicles with a diameter of 30-100 nm derived from a variety of eukaryocytes. The cargo of exosomes includes proteins, lipids, nucleic acids, and substances of the cells from which they originate. They can transfer functional cargo to neighboring and distal cells, therefore contributing to intercellular communication in both physiological and pathological processes. In recent years, it was shown that exosomes in several neurodegenerative diseases are closely related to the transmission of disease-related misfolded proteins (such as α-synuclein, tau, amyloid β-protein, etc). These proteins are transported by exosomes, thus promoting the propagation to unaffected cells or areas and accelerating the progression of neurodegenerative diseases. This review focuses on the origin and composition, biological synthesis, secretion, function of exosomes, as well as their roles in the pathogenesis and progression of neurodegenerative diseases. In addition, we also discuss that exosomes can serve as biomarkers and drug delivery vehicles, and play a role in the diagnosis and treatment of neurodegenerative diseases.

Exosomes in the Pathogenesis of Rheumatoid Arthritis / 中国医学科学院学报

Exosomes are 30-100 nm vesicles secreted from almost all types of cells.They contain various molecular constituents,including proteins,lipids,and RNA.As important mediators of cell-to-cell communication,exosomes are involved in a variety of physiological and pathological processes such as inflammatory reaction,cell proliferation and differentiation,tissue repair,immune signal transduction,and stress response.Exosomes can regulate and maintain the initiation and progression of many autoimmune diseases,especially rheumatoid arthritis.Meanwhile,exosomes may be a new biomarker for the diagnosis of rheumatoid arthritis and a potential treatment vector for this disease.

El Sexting a través del discurso de adolescentes españoles / Sexting through the Spanish adolescent discourse

Resumen El objetivo de esta investigación fue indagar en las prácticas de sexting en un grupo de adolescentes españoles y contribuir así a su mejor comprensión a través de una investigación novedosa en España. Se pretende descubrir los conocimientos que los/las adolescentes tienen sobre el fenómeno del sexting y averiguar en sus comportamientos y motivaciones hacia el sexting, revelando si perciben consecuencias de este tipo de conducta. A través de la aproximación cualitativa se realizaron ocho focus group, conformados por 89 adolescentes, que tenían entre 14 y 18 años. Entre los resultados se destacan el desconocimiento del concepto de sexting, llegando incluso a confundirlo con el acoso y el chantaje. Aunque solo un quinto de los y las participantes reconoce practicar sexting, admiten que es una práctica muy frecuente entre la gente de sus edades, especialmente entre las chicas. Los motivos que aluden para justificar su participación en comportamientos de sexting son fundamentalmente de carácter sexual, mientras que señalan que la gente de sus edades sextea por diversión, aburrimiento o por estar de moda. Asimismo, reconocen que hay ciertos riesgos derivados de las conductas de sexting, que afectan más negativamente a las chicas. Además, la gran parte de los/las jóvenes son conocedores de situaciones entre sus iguales de coacciones y chantajes para enviar contenido erótico-sexual, así como de otras realidades como el ciberacoso y el ciberstalking.

Abstract The objective of this research was to investigate the practices of sexting in a group of Spanish adolescents and thus contribute to their better understanding through a novel research in Spain. The aim is to discover the knowledge that adolescents have about the phenomenon of sexting and to investigate their behaviors and motivations towards sexting, revealing if they perceive consequences of this type of behavior. Through the qualitative approach, eight focus groups were made, composed of 89 adolescents, from 14 to 18 years old. Among the results they emphasize the ignorance of the concept of sexting, even confusing it with harassment and blackmail. Although only a fifth of the participants recognize practicing sexting, they admit that it is a very frequent practice among people of their age, especially among girls. The reasons they allude to justify their participation in sexting behaviors are fundamentally sexual in nature, while they point out that people of their age carry out sexting behavior for fun, boredom or for being fashionable. They also recognize there are certain risks derived from sexting behaviors, which affect girls more negatively. In addition, most young people are aware of situations of coercion and blackmail among their peers to send erotic-sexual content, as well as other realities such as cyberbullying and cyberstalking.

Priming stem cells with protein kinase C activator enhances early stem cell-chondrocyte interaction by increasing adhesion molecules

BACKGROUND: Osteoarthritis (OA) can be defined as degradation of articular cartilage of the joint, and is the most common degenerative disease. To regenerate the damaged cartilage, different experimental approaches including stem cell therapy have been tried. One of the major limitations of stem cell therapy is the poor post-transplantation survival of the stem cells. Anoikis, where insufficient matrix support and adhesion to extracellular matrix causes apoptotic cell death, is one of the main causes of the low post-transplantation survival rate of stem cells. Therefore, enhancing the initial interaction of the transplanted stem cells with chondrocytes could improve the therapeutic efficacy of stem cell therapy for OA. Previously, protein kinase C activator phorbol 12-myristate 13-acetate (PMA)- induced increase of mesenchymal stem cell adhesion via activation of focal adhesion kinase (FAK) has been reported. In the present study, we examine the effect PMA on the adipose-derived stem cells (ADSCs) adhesion and spreading to culture substrates, and further on the initial interaction between ADSC and chondrocytes. RESULTS: PMA treatment increased the initial adhesion of ADSC to culture substrate and cellular spreading with increased expression of adhesion molecules, such as FAK, vinculin, talin, and paxillin, at both RNA and protein level. Priming of ADSC with PMA increased the number of ADSCs attached to confluent layer of cultured chondrocytes compared to that of untreated ADSCs at early time point (4 h after seeding). CONCLUSION: Taken together, the results of this study suggest that priming ADSCs with PMA can increase the initial interaction with chondrocytes, and this proof of concept can be used to develop a non-invasive therapeutic approach for treating OA. It may also accelerate the regeneration process so that it can relieve the accompanied pain faster in OA patients. Further in vivo studies examining the therapeutic effect of PMA pretreatment of ADSCs for articular cartilage damage are required.

Acebutolol, a Cardioselective Beta Blocker, Promotes Glucose Uptake in Diabetic Model Cells by Inhibiting JNK-JIP1 Interaction

The phosphorylation of JNK is known to induce insulin resistance in insulin target tissues. The inhibition of JNK-JIP1 interaction, which interferes JNK phosphorylation, becomes a potential target for drug development of type 2 diabetes. To discover the inhibitors of JNK-JIP1 interaction, we screened out 30 candidates from 4320 compound library with In Cell Interaction Trap method. The candidates were further confirmed and narrowed down to five compounds using the FRET method in a model cell. Among those five compounds, Acebutolol showed notable inhibition of JNK phosphorylation and elevation of glucose uptake in diabetic models of adipocyte and liver cell. Structural computation showed that the binding affinity of Acebutolol on the JNK-JIP1 interaction site was comparable to the known inhibitor, BI-78D3. Our results suggest that Acebutolol, an FDA-approved beta blocker for hypertension therapy, could have a new repurposed effect on type 2 diabetes elevating glucose uptake process by inhibiting JNK-JIP1 interaction.