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1.
J Transl Med ; 22(1): 546, 2024 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-38849907

RESUMO

BACKGROUND: The pathogenesis of thyroid-associated orbitopathy (TAO) remains incompletely understand. The interaction between immunocytes and orbital fibroblasts (OFs) play a critical role in orbital inflammatory and fibrosis. Accumulating reports indicate that a significant portion of plasma exosomes (Pla-Exos) are derived from immune cells; however, their impact upon OFs function is unclear. METHODS: OFs were primary cultured from inactive TAO patients. Exosomes isolated from plasma samples of patients with active TAO and healthy controls (HCs) were utilized for functional and RNA cargo analysis. Functional analysis in thymocyte differentiation antigen-1+ (Thy-1+) OFs measured expression of inflammatory and fibrotic markers (mRNAs and proteins) and cell activity in response to Pla-Exos. RNA cargo analysis was performed by RNA sequencing and RT-qPCR. Thy-1+ OFs were transfected with miR-144-3p mimics/inhibitors to evaluate its regulation of inflammation, fibrosis, and proliferation. RESULTS: Pla-Exos derived from active TAO patients (Pla-ExosTAO-A) induced stronger production of inflammatory cytokines and hyaluronic acid (HA) in Thy-1+ OFs while inhibiting their proliferation. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and single sample gene set enrichment analysis (ssGSEA) suggested that the difference in mRNA expression levels between Pla-ExosTAO-A and Pla-ExosHC was closely related to immune cells. Differential expression analysis revealed that 62 upregulated and 45 downregulated miRNAs in Pla-ExosTAO-A, with the elevation of miR-144-3p in both Pla-Exos and PBMCs in active TAO group. KEGG analysis revealed that the target genes of differentially expressed miRNA and miR-144-3p enriched in immune-related signaling pathways. Overexpression of the miR-144-3p mimic significantly upregulated the secretion of inflammatory cytokines and HA in Thy-1+ OFs while inhibiting their proliferation. CONCLUSION: Pla-Exos derived from patients with active TAO were immune-active, which may be a long-term stimulus casual for inflammatory and fibrotic progression of TAO. Our finding suggests that Pla-Exos could be used as biomarkers or treatment targets in TAO patients.


Assuntos
Exossomos , Fibroblastos , Fibrose , Oftalmopatia de Graves , Inflamação , MicroRNAs , Órbita , Humanos , Exossomos/metabolismo , Oftalmopatia de Graves/patologia , Oftalmopatia de Graves/sangue , Oftalmopatia de Graves/genética , MicroRNAs/genética , MicroRNAs/metabolismo , MicroRNAs/sangue , Fibroblastos/metabolismo , Fibroblastos/patologia , Órbita/patologia , Inflamação/patologia , Feminino , Masculino , Proliferação de Células , Pessoa de Meia-Idade , Adulto , Ácido Hialurônico/sangue , Ácido Hialurônico/metabolismo , Citocinas/metabolismo , Antígenos Thy-1/metabolismo
2.
BMC Cancer ; 24(1): 671, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38824581

RESUMO

BACKGROUND: The role of novel circular RNAs (circRNAs) in colorectal cancer (CRC) remains to be determined. This study aimed to identify a novel circRNA involved in CRC pathogenesis, assess its diagnostic value, and construct a regulatory network. METHODS: Differential expression analysis was conducted using circRNA datasets to screen for differentially expressed circRNAs. The expression of selected circRNAs was validated in external datasets and clinical samples. Diagnostic value of plasma circRNA levels in CRC was assessed. A competing endogenous RNA (ceRNA) network was constructed for the circRNA using TCGA dataset. RESULTS: Analysis of datasets revealed that hsa_circ_101303 was significantly overexpressed in CRC tissues compared to normal tissues. The upregulation of hsa_circ_101303 in CRC tissues was further confirmed through the GSE138589 dataset and clinical samples. High expression of hsa_circ_101303 was associated with advanced N stage, M stage, and tumor stage in CRC. Plasma levels of hsa_circ_101303 were markedly elevated in CRC patients and exhibited moderate diagnostic ability for CRC (AUC = 0.738). The host gene of hsa_circ_101303 was also found to be related to the TNM stage of CRC. Nine miRNAs were identified as target miRNAs for hsa_circ_101303, and 27 genes were identified as targets of these miRNAs. Subsequently, a ceRNA network for hsa_circ_101303 was constructed to illustrate the interactions between the nine miRNAs and 27 genes. CONCLUSIONS: The study identifies hsa_circ_101303 as a highly expressed circRNA in CRC, which is associated with the progression of the disease. Plasma levels of hsa_circ_101303 show promising diagnostic potential for CRC. The ceRNA network for hsa_circ_101303 provides valuable insights into the regulatory mechanisms underlying CRC.


Assuntos
Biomarcadores Tumorais , Neoplasias Colorretais , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , MicroRNAs , RNA Circular , Humanos , Neoplasias Colorretais/genética , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/sangue , Neoplasias Colorretais/patologia , RNA Circular/genética , RNA Circular/sangue , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/sangue , Masculino , Feminino , MicroRNAs/genética , MicroRNAs/sangue , Pessoa de Meia-Idade , Perfilação da Expressão Gênica , Estadiamento de Neoplasias
3.
Anal Chim Acta ; 1315: 342822, 2024 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-38879216

RESUMO

In this study, a novel electrochemiluminescence (ECL) biosensor was developed to detect microRNA-21 (miRNA-21) with high sensitivity by leveraging the combined mechanisms of resonance energy transfer (RET) and surface plasmon coupling (SPC). Initially, the glassy carbon electrode (GCE) were coated with Cu-Zn-In-S quantum dots (CZIS QDs), known for their defect-related emission suitable for ECL sensing. Subsequently, a hairpin DNA H3 with gold nanoparticles (Au NPs) attached at the end was modified over the surface of the quantum dots. The Au NPs could effectively quench the ECL signals of CZIS QDs via RET. Further, a significant amount of report DNA was generated through the action of a 3D DNA walker. When the report DNA opened H3-Au NPs, the hairpin structure experienced a conformational change to a linear shape, increasing the gap between the CZIS QDs and the Au NPs. Consequently, the localized surface plasmon resonance ECL (LSPR-ECL) effect replaced ECL resonance energy transfer (ECL-RET). Moreover, the report DNA was released following the addition of H4-Au NPs, resulting in the formation of Au dimers and a surface plasma-coupled ECL (SPC-ECL) effect that enhanced the ECL intensity to 6.97-fold. The integration of new ECL-RET and SPC-ECL biosensor accurately quantified miRNA-21 concentrations from 10-8 M to 10-16 M with a limit of detection (LOD) of 0.08 fM, as well as successfully applied to validate human serum samples.


Assuntos
Técnicas Biossensoriais , DNA , Técnicas Eletroquímicas , Medições Luminescentes , MicroRNAs , Pontos Quânticos , Ressonância de Plasmônio de Superfície , MicroRNAs/análise , MicroRNAs/sangue , Humanos , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , DNA/química , Pontos Quânticos/química , Ressonância de Plasmônio de Superfície/métodos , Medições Luminescentes/métodos , Ouro/química , Limite de Detecção , Transferência de Energia , Nanopartículas Metálicas/química
4.
Sci Rep ; 14(1): 13702, 2024 06 14.
Artigo em Inglês | MEDLINE | ID: mdl-38871789

RESUMO

Coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Recent research has demonstrated how epigenetic mechanisms regulate the host-virus interactions in COVID-19. It has also shown that microRNAs (miRNAs) are one of the three fundamental mechanisms of the epigenetic regulation of gene expression and play an important role in viral infections. A pilot study published by our research group identified, through next-generation sequencing (NGS), that miR-4433b-5p, miR-320b, and miR-16-2-3p are differentially expressed between patients with COVID-19 and controls. Thus, the objectives of this study were to validate the expression of these miRNAs using quantitative real-time polymerase chain reaction (qRT-PCR) and to perform in silico analyses. Patients with COVID-19 (n = 90) and healthy volunteers (n = 40) were recruited. MiRNAs were extracted from plasma samples and validated using qRT-PCR. In addition, in silico analyses were performed using mirPath v.3 software. MiR-320b was the only miRNA upregulated in the case group com-pared to the control group. The in silico analyses indicated the role of miR-320b in the regulation of the KITLG gene and consequently in the inflammatory process. This study confirmed that miR-320b can distinguish patients with COVID-19 from control participants; however, further research is needed to determine whether this miRNA can be used as a target or a biomarker.


Assuntos
COVID-19 , MicroRNAs , SARS-CoV-2 , Humanos , COVID-19/genética , COVID-19/sangue , COVID-19/virologia , MicroRNAs/sangue , MicroRNAs/genética , Masculino , Feminino , SARS-CoV-2/genética , Pessoa de Meia-Idade , Adulto , Idoso , Estudos de Casos e Controles
5.
Parasit Vectors ; 17(1): 258, 2024 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-38872227

RESUMO

BACKGROUND: Dirofilaria immitis, commonly known as heartworm (HW), is a parasitic nematode transmitted by various mosquito species, leading to heartworm disease (HWD) in dogs. Diagnosis of HW typically involves antigen or microfilariae detection, or visualization of adult worms through imaging or post mortem examination. Polymerase chain reaction (PCR) and micro RNA (miRNA) detection have been explored for HW diagnosis. METHODS: Three dogs, previously experimentally infected with HW, underwent blood sampling every 4 weeks for 7 months. Samples were assessed for antigen presence after heat treatment, PCR amplification, and microfilaria examination using Giemsa-stained thick smears. Additionally, whole blood aliquots underwent miRNA deep sequencing and bioinformatic analysis. RESULTS: Heartworm antigen was detectable after heat treatment at 20 weeks post-inoculation and via PCR at 24 weeks, with microfilariae observed in peripheral blood smears at 28 weeks. However, deep miRNA sequencing revealed that the miRNA candidate sequences are not consistently expressed before 28 weeks of infection. CONCLUSIONS: While ancillary molecular methods such as PCR and miRNA sequencing may be less effective than antigen detection for detecting immature larval stages in an early stage of infection, our experimental findings demonstrate that circulating miRNAs can still be detected in 28 weeks post-infection.


Assuntos
Dirofilaria immitis , Dirofilariose , Doenças do Cão , MicroRNAs , Animais , Dirofilaria immitis/genética , Dirofilaria immitis/isolamento & purificação , Cães , Dirofilariose/diagnóstico , Dirofilariose/parasitologia , MicroRNAs/sangue , MicroRNAs/genética , Doenças do Cão/parasitologia , Doenças do Cão/diagnóstico , Antígenos de Helmintos/sangue , Antígenos de Helmintos/genética , Diagnóstico Precoce , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , Microfilárias/isolamento & purificação , Microfilárias/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos
6.
Sci Rep ; 14(1): 13573, 2024 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-38866792

RESUMO

Angiotensin converting enzyme 2 (ACE2) serves as the primary receptor for the SARS-CoV-2 virus and has implications for the functioning of the cardiovascular system. Based on our previously published bioinformatic analysis, in this study we aimed to analyze the diagnostic and predictive utility of miRNAs (miR-10b-5p, miR-124-3p, miR-200b-3p, miR-26b-5p, miR-302c-5p) identified as top regulators of ACE2 network with potential to affect cardiomyocytes and cardiovascular system in patients with COVID-19. The expression of miRNAs was determined through qRT-PCR in a cohort of 79 hospitalized COVID-19 patients as well as 32 healthy volunteers. Blood samples and clinical data of COVID-19 patients were collected at admission, 7-days and 21-days after admission. We also performed SHAP analysis of clinical data and miRNAs target predictions and advanced enrichment analyses. Low expression of miR-200b-3p at the seventh day of admission is indicative of predictive value in determining the length of hospital stay and/or the likelihood of mortality, as shown in ROC curve analysis with an AUC of 0.730 and a p-value of 0.002. MiR-26b-5p expression levels in COVID-19 patients were lower at the baseline, 7 and 21-days of admission compared to the healthy controls (P < 0.0001). Similarly, miR-10b-5p expression levels were lower at the baseline and 21-days post admission (P = 0.001). The opposite situation was observed in miR-124-3p and miR-302c-5p. Enrichment analysis showed influence of analyzed miRNAs on IL-2 signaling pathway and multiple cardiovascular diseases through COVID-19-related targets. Moreover, the COVID-19-related genes regulated by miR-200b-3p were linked to T cell protein tyrosine phosphatase and the HIF-1 transcriptional activity in hypoxia. Analysis focused on COVID-19 associated genes showed that all analyzed miRNAs are strongly affecting disease pathways related to CVDs which could be explained by their strong interaction with the ACE2 network.


Assuntos
Enzima de Conversão de Angiotensina 2 , COVID-19 , MicroRNAs , Humanos , COVID-19/sangue , COVID-19/genética , COVID-19/virologia , Masculino , Feminino , Pessoa de Meia-Idade , Enzima de Conversão de Angiotensina 2/genética , Enzima de Conversão de Angiotensina 2/sangue , Enzima de Conversão de Angiotensina 2/metabolismo , Idoso , MicroRNAs/sangue , MicroRNAs/genética , SARS-CoV-2/genética , Redes Reguladoras de Genes , MicroRNA Circulante/sangue , MicroRNA Circulante/genética , Adulto
7.
Sci Rep ; 14(1): 13560, 2024 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-38866905

RESUMO

L1CAM-positive extracellular vesicles (L1EV) are an emerging biomarker that may better reflect ongoing neuronal damage than other blood-based biomarkers. The physiological roles and regulation of L1EVs and their small RNA cargoes following stroke is unknown. We sought to characterize L1EV small RNAs following stroke and assess L1EV RNA signatures for diagnosing stroke using weighted gene co-expression network analysis and random forest (RF) machine learning algorithms. Interestingly, small RNA sequencing of plasma L1EVs from patients with stroke and control patients (n = 28) identified micro(mi)RNAs known to be enriched in the brain. Weighted gene co-expression network analysis (WGCNA) revealed small RNA transcript modules correlated to diagnosis, initial NIH stroke scale, and age. L1EV RNA signatures associated with the diagnosis of AIS were derived from WGCNA and RF classification. These small RNA signatures demonstrated a high degree of accuracy in the diagnosis of AIS with an area under the curve (AUC) of the signatures ranging from 0.833 to 0.932. Further work is necessary to understand the role of small RNA L1EV cargoes in the response to brain injury, however, this study supports the utility of L1EV small RNA signatures as a biomarker of stroke.


Assuntos
Biomarcadores , Vesículas Extracelulares , AVC Isquêmico , Molécula L1 de Adesão de Célula Nervosa , Humanos , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/genética , Masculino , AVC Isquêmico/genética , AVC Isquêmico/metabolismo , AVC Isquêmico/sangue , AVC Isquêmico/diagnóstico , Molécula L1 de Adesão de Célula Nervosa/genética , Molécula L1 de Adesão de Célula Nervosa/metabolismo , Feminino , Idoso , Biomarcadores/sangue , Pessoa de Meia-Idade , Aprendizado de Máquina , MicroRNAs/genética , MicroRNAs/sangue , MicroRNAs/metabolismo
8.
Parasit Vectors ; 17(1): 256, 2024 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-38867315

RESUMO

BACKGROUND: Human toxocariasis is a neglected parasitic disease characterised by the syndromes visceral, cerebral, and ocular larva migrans. This disease is caused by the migrating larvae of Toxocara roundworms from dogs and cats, affecting 1.4 billion people globally. Via extracellular vesicles (EVs), microRNAs have been demonstrated to play roles in host-parasite interactions and proposed as circulating biomarkers for the diagnosis and follow-up of parasitic diseases. METHODS: Small RNA-seq was conducted to identify miRNAs in the infective larvae of T. canis and plasma EV-containing preparations of infected BALB/c mice. Differential expression analysis and target prediction were performed to indicate miRNAs involved in host-parasite interactions and miRNAs associated with visceral and/or cerebral larva migrans in the infected mice. Quantitative real-time polymerase chain reaction (PCR) was used to amplify circulating miRNAs from the infected mice. RESULTS: This study reports host and parasite miRNAs in the plasma of BALB/c mice with visceral and cerebral larva migrans and demonstrates the alterations of these miRNAs during the migration of larvae from the livers through the lungs and to the brains of infected mice. After filtering unspecific changes in an irrelevant control, T. canis-derived miRNAs and T. canis infection-induced differential miRNAs are predicted to modulate genes consistently involved in mitogen-activated protein kinase (MAPK) signalling and pathways regulating axon guidance and pluripotency of stem in the infected mice with visceral and cerebral larva migrans. For these plasma circulating miRNAs predicted to be involved in host-parasite crosstalk, two murine miRNAs (miR-26b-5p and miR-122-5p) are experimentally verified to be responsive to larva migrans and represent circulating biomarker candidates for visceral and cerebral toxocariasis in BALB/c mice. CONCLUSIONS: Our findings provide novel insights into the crosstalk of T. canis and the mammalian host via plasma circulating miRNAs, and prime agents and indicators for visceral and cerebral larva migrans. A deep understanding of these aspects will underpin the diagnosis and control of toxocariasis in humans and animals.


Assuntos
MicroRNA Circulante , Camundongos Endogâmicos BALB C , Toxocara canis , Toxocaríase , Animais , Toxocara canis/genética , Toxocara canis/fisiologia , Camundongos , Toxocaríase/parasitologia , Toxocaríase/sangue , MicroRNA Circulante/sangue , MicroRNA Circulante/genética , Interações Hospedeiro-Parasita , Larva Migrans Visceral/parasitologia , Larva Migrans Visceral/sangue , Feminino , Larva Migrans/parasitologia , Larva Migrans/sangue , Larva/genética , Cães , MicroRNAs/sangue , MicroRNAs/genética , Biomarcadores/sangue , Encéfalo/parasitologia
9.
Hum Genomics ; 18(1): 64, 2024 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-38872198

RESUMO

BACKGROUND: The 22q11.2 deletion syndrome (22q11.2DS) is a microdeletion syndrome with highly variable phenotypic manifestations, even though most patients present the typical 3 Mb microdeletion, usually affecting the same ~ 106 genes. One of the genes affected by this deletion is DGCR8, which plays a crucial role in miRNA biogenesis. Therefore, the haploinsufficiency of DGCR8 due to this microdeletion can alter the modulation of the expression of several miRNAs involved in a range of biological processes. RESULTS: In this study, we used next-generation sequencing to evaluate the miRNAs profiles in the peripheral blood of 12 individuals with typical 22q11DS compared to 12 healthy matched controls. We used the DESeq2 package for differential gene expression analysis and the DIANA-miTED dataset to verify the expression of differentially expressed miRNAs in other tissues. We used miRWalk to predict the target genes of differentially expressed miRNAs. Here, we described two differentially expressed miRNAs in patients compared to controls: hsa-miR-1304-3p, located outside the 22q11.2 region, upregulated in patients, and hsa-miR-185-5p, located in the 22q11.2 region, which showed downregulation. Expression of miR-185-5p is observed in tissues frequently affected in patients with 22q11DS, and previous studies have reported its downregulation in individuals with 22q11DS. hsa-miR-1304-3p has low expression in blood and, thus, needs more validation, though using a sensitive technology allowed us to identify differences in expression between patients and controls. CONCLUSIONS: Thus, lower expression of miR-185-5p can be related to the 22q11.2 deletion and DGCR8 haploinsufficiency, leading to phenotypic consequences in 22q11.2DS patients, while higher expression of hsa-miR-1304-3p might be related to individual genomic variances due to the heterogeneous background of the Brazilian population.


Assuntos
Síndrome de DiGeorge , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , MicroRNAs , Humanos , MicroRNAs/genética , MicroRNAs/sangue , Masculino , Feminino , Síndrome de DiGeorge/genética , Síndrome de DiGeorge/patologia , Criança , Adolescente , Adulto , Estudos de Casos e Controles , Proteínas de Ligação a RNA/genética , Regulação da Expressão Gênica/genética , Haploinsuficiência/genética , Adulto Jovem
10.
Eur J Sport Sci ; 24(6): 766-776, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38874986

RESUMO

A sedentary lifestyle and Olympic participation are contrary risk factors for global mortality and incidence of cancer and cardiovascular disease. Extracellular vesicle miRNAs have been described to respond to exercise. No molecular characterization of young male sedentary people versus athletes is available; so, our aim was to identify the extracellular vesicle miRNA profile of chronically trained young endurance and resistance male athletes compared to their sedentary counterparts. A descriptive case-control design was used with 16 sedentary young men, 16 Olympic male endurance athletes, and 16 Olympic male resistance athletes. Next-generation sequencing and RT-qPCR and external and internal validation were performed in order to analyze extracellular vesicle miRNA profiles. Endurance and resistance athletes had significant lower levels of miR-16-5p, miR-19a-3p, and miR-451a compared to sedentary people. Taking all together, exercise-trained miRNA profile in extracellular vesicles provides a differential signature of athletes irrespective of the type of exercise compared to sedentary people. Besides, miR-25-3p levels were specifically lower in endurance athletes which defines its role as a specific responder in this type of athletes. In silico analysis of this profile suggests a role in adaptive energy metabolism in this context that needs to be experimentally validated. Therefore, this study provides for the first time basal levels of circulating miRNA in extracellular vesicles emerge as relevant players in intertissue communication in response to chronic exercise exposure in young elite male athletes.


Assuntos
Atletas , Vesículas Extracelulares , Sequenciamento de Nucleotídeos em Larga Escala , MicroRNAs , Comportamento Sedentário , Humanos , Masculino , MicroRNAs/sangue , Vesículas Extracelulares/metabolismo , Estudos de Casos e Controles , Adulto Jovem , Resistência Física , Adolescente
11.
PLoS One ; 19(6): e0305042, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38861512

RESUMO

Multiple sclerosis (MS) and glioblastoma (GBM) are CNS diseases in whose development and progression immune privilege is intimately important, but in a relatively opposite manner. Maintenance and strengthening of immune privilege have been shown to be an important mechanism in glioblastoma immune evasion, while the breakdown of immune privilege leads to MS initiation and exacerbation. We hypothesize that molecular signaling pathways can be oppositely regulated in peripheral blood CD8+ T cells of MS and glioblastoma patients at a transcriptional level. We analyzed publicly available data of the peripheral blood CD8+ T cell MS vs. control (MSvsCTRL) and GBM vs. control (GBMvsCTRL) differentially expressed gene (DEG) contrasts with Qiagen's Ingenuity pathway analysis software (IPA). We have identified sphingolipid signaling pathway which was significantly downregulated in the GBMvsCTRL and upregulated in the MSvsCTRL. As the pathway is important for the CD8+ T lymphocytes CNS infiltration, this result is in line with our previously stated hypothesis. Comparing publicly available lists of differentially expressed serum exosomal miRNAs from MSvsCTRL and GBMvsCTRL contrasts, we have identified that hsa-miR-182-5p has the greatest potential effect on sphingolipid signaling regarding the number of regulated DEGs in the GBMvsCTRL contrast, while not being able to find any relevant potential sphingolipid signaling target transcripts in the MSvsCTRL contrast. We conclude that the sphingolipid signaling pathway is a top oppositely regulated pathway in peripheral blood CD8+ T cells from GBM and MS, and might be crucial for the differences in CNS immune privilege maintenance of investigated diseases, but further experimental research is necessary.


Assuntos
Linfócitos T CD8-Positivos , Glioblastoma , MicroRNAs , Esclerose Múltipla , Transdução de Sinais , Esfingolipídeos , Transcriptoma , Humanos , Glioblastoma/imunologia , Glioblastoma/genética , Glioblastoma/sangue , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Esfingolipídeos/sangue , Esfingolipídeos/metabolismo , Esclerose Múltipla/imunologia , Esclerose Múltipla/sangue , Esclerose Múltipla/genética , MicroRNAs/genética , MicroRNAs/sangue , Perfilação da Expressão Gênica , Neoplasias Encefálicas/imunologia , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/sangue , Neoplasias Encefálicas/patologia , Regulação Neoplásica da Expressão Gênica
12.
Int Heart J ; 65(3): 498-505, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38825494

RESUMO

This study aimed to explore the expression of long non-coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) in patients with acute myocardial infarction (AMI) and its inflammatory regulation mechanism through miR-211/interleukin 10 (IL-10) axis.A total of 75 participants were enrolled in this study: 25 healthy people in the control group, 25 patients with stable angina pectoris (SAP) in the SAP group, and 25 patients with AMI in the AMI group. Real-time qPCR was used to detect mRNA expression levels of NEAT1, miR-211, and IL-10. The interaction between miR-211, NEAT1, and IL-10 was confirmed by dual-luciferase reporter assay, and protein expression was detected using western blot.High expression of NEAT1 in peripheral blood mononuclear cells (PBMCs) of patients with AMI was negatively related to serum creatine kinase-MB (CK-MB), cardiac troponin I (cTnI), tumor necrosis factor-α (TNF-α), IL-6, and IL-1ß and was positively correlated with left ventricular ejection fraction (LVEF). In THP-1 cells, miR-211 was confirmed to target and inhibit IL-10 expression. NEAT1 knockdown and miR-211-mimic markedly decreased IL-10 protein levels, whereas anti-miR-211 markedly increased IL-10 protein levels. Importantly, miR-211 level was negatively related to NEAT1 and IL-10 levels, whereas IL-10 level was positively related to the level of NEAT1 expression in PBMCs of patients with AMI.LncRNA NEAT1 was highly expressed in PBMCs of patients with AMI, and NEAT1 suppressed inflammation via miR-211/IL-10 axis in PBMCs of patients with AMI.


Assuntos
Interleucina-10 , Leucócitos Mononucleares , MicroRNAs , Infarto do Miocárdio , RNA Longo não Codificante , Humanos , RNA Longo não Codificante/genética , RNA Longo não Codificante/sangue , MicroRNAs/sangue , MicroRNAs/genética , Interleucina-10/sangue , Interleucina-10/metabolismo , Infarto do Miocárdio/sangue , Infarto do Miocárdio/genética , Infarto do Miocárdio/metabolismo , Leucócitos Mononucleares/metabolismo , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Inflamação/genética , Inflamação/sangue , Inflamação/metabolismo , Estudos de Casos e Controles
13.
Genomics ; 116(4): 110875, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38849018

RESUMO

Exploration of a stably expressed gene as a reference is critical for the accurate evaluation of miRNAs isolated from small extracellular vesicles (sEVs). In this study, we analyzed small RNA sequencing on plasma sEV miRNAs in the training dataset (n = 104) and found that miR-140-3p was the most stably expressed candidate reference for sEV miRNAs. We further demonstrated that miR-140-3p expressed most stably in the validation cohort (n = 46) when compared to two other reference miRNAs, miR-451a and miR-1228-3p, and the commonly-used miRNA reference U6. Finally, we compared the capability of miR-140-3p and U6 as the internal reference for sEV miRNA expression by evaluating key miRNAs expression in lung cancer patients and found that miR-140-3p was more suitable as a sEV miRNA reference gene. Taken together, our data indicated miR-140-3p as a stable internal reference miRNA of plasma sEVs to evaluate miRNA expression profiles in lung cancer patients.


Assuntos
Vesículas Extracelulares , Neoplasias Pulmonares , MicroRNAs , Humanos , MicroRNAs/sangue , MicroRNAs/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/sangue , Vesículas Extracelulares/genética , Vesículas Extracelulares/metabolismo , Feminino , Masculino , Padrões de Referência , Reação em Cadeia da Polimerase em Tempo Real/normas , Pessoa de Meia-Idade , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/genética
14.
Biochem Med (Zagreb) ; 34(2): 020501, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38882585

RESUMO

Microribonucleic acids (miRNAs) have emerged as a new category of biomarkers for many human diseases like cancer, cardiovascular and neurodegenerative disorders. MicroRNAs can be detected in various body fluids including blood, urine and cerebrospinal fluid. However, the literature contains conflicting results for circulating miRNAs, which is the main barrier to using miRNAs as non-invasive biomarkers. This variability in results is largely due to differences between studies in sample processing methodology, miRNA quantification and result normalization. The purpose of this review is to describe the various preanalytical, analytical and postanalytical factors that can impact miRNA detection accuracy and to propose recommendations for the standardization of circulating miRNAs measurement.


Assuntos
MicroRNA Circulante , Humanos , MicroRNA Circulante/sangue , Biomarcadores/sangue , Fase Pré-Analítica , MicroRNAs/sangue
15.
J Am Heart Assoc ; 13(12): e033674, 2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38860398

RESUMO

BACKGROUND: Extracellular microRNAs (miRNAs) are a class of noncoding RNAs that remain stable in the extracellular milieu, where they contribute to various physiological and pathological processes by facilitating intercellular signaling. Previous studies have reported associations between miRNAs and cardiovascular diseases (CVDs); however, the plasma miRNA signatures of CVD and its risk factors have not been fully elucidated at the population level. METHODS AND RESULTS: Plasma miRNA levels were measured in 4440 FHS (Framingham Heart Study) participants. Linear regression analyses were conducted to test the cross-sectional associations of each miRNA with 8 CVD risk factors. Prospective analyses of the associations of miRNAs with new-onset obesity, hypertension, type 2 diabetes, CVD, and all-cause mortality were conducted using proportional hazards regression. Replication was carried out in 1999 RS (Rotterdam Study) participants. Pathway enrichment analyses were conducted and target genes were predicted for miRNAs associated with ≥5 risk factors in the FHS. In the FHS, 6 miRNAs (miR-193b-3p, miR-122-5p, miR-365a-3p, miR-194-5p, miR-192-5p, and miR-193a-5p) were associated with ≥5 risk factors. This miRNA signature was enriched for pathways associated with CVD and several genes annotated to these pathways were predicted targets of the identified miRNAs. Furthermore, miR-193b-3p, miR-194-5p, and miR-193a-5p were each associated with ≥2 risk factors in the RS. Prospective analysis revealed 8 miRNAs associated with all-cause mortality in the FHS. CONCLUSIONS: These findings highlight associations between miRNAs and CVD risk factors that may provide valuable insights into the underlying pathogenesis of CVD.


Assuntos
Doenças Cardiovasculares , Fatores de Risco de Doenças Cardíacas , MicroRNAs , Humanos , Masculino , Doenças Cardiovasculares/genética , Doenças Cardiovasculares/sangue , Doenças Cardiovasculares/mortalidade , Feminino , Pessoa de Meia-Idade , Idoso , MicroRNAs/sangue , MicroRNAs/genética , Estudos Prospectivos , Estudos Transversais , Medição de Risco , MicroRNA Circulante/sangue , MicroRNA Circulante/genética , Fatores de Risco , Biomarcadores/sangue , Fatores Etários
16.
Clin Cardiol ; 47(6): e24274, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38884329

RESUMO

BACKGROUND: Atherosclerotic cardiovascular disease (ASCVD) is a group of clinical diseases based on pathology of atherosclerosis that is the leading cause of mortality worldwide. There is a bidirectional interaction between ASCVD and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Alterations in circulating miRNAs levels are involved in the development of ASCVD in patients infected with SARS-CoV-2, however, the correlation between ASCVD co-infection with SARS-CoV-2 and alterations of cardiac-specific miRNAs is not well understood. HYPOTHESIS: The circulating miR-146a and miR-27a are involved in bidirectional interactions between ASCVD and SARS-CoV-2 infections. METHODS: Circulating miR-146a and miR-27a levels were measured in serum and PBMCs deriving from ASCVD patients and controls after SARS-CoV-2 infection by qRT-PCR analysis. The levels of neutralizing antibodies-resistant SARS-CoV-2 in human serum was determined by competitive magnetic particle chemiluminescence method. Interleukin (IL)-6 levels were detected by automatic biochemical analyzer using electrochemiluminescence. RESULTS: Significant downregulation of circulating miR-146a and upregulation of miR-27a in ASCVD patients after infection with SARS-CoV-2 compared with controls were observed, among which the alterations were more evident in ASCVD patients comorbid with hyperlipidemia and diabetes mellitus. Consistently, correlation analysis revealed that serum miR-146a and miR-27a levels were associated with the levels of lipids and glucose, inflammatory response, and immune function in ASCVD patients. Remarkably, SARS-CoV-2 S protein RBD stimulation of PBMCs derived from both ASCVD and controls significantly downregulated miR-146a, upregulated miR-27a expression levels, and promoted IL-6 release in vitro. CONCLUSIONS: The circulating miR-146a and miR-27a are involved in metabolism, inflammation, and immune levels in patients with ASCVD after SARS-CoV-2 infection, laying the foundation for the development of strategies to prevent the risk of SARS-CoV-2 infection in ASCVD patients.


Assuntos
Aterosclerose , COVID-19 , MicroRNAs , SARS-CoV-2 , Humanos , COVID-19/sangue , COVID-19/imunologia , COVID-19/complicações , MicroRNAs/sangue , Masculino , Feminino , Pessoa de Meia-Idade , Aterosclerose/sangue , Aterosclerose/epidemiologia , Idoso , Biomarcadores/sangue , MicroRNA Circulante/sangue
17.
J Cell Mol Med ; 28(11): e18370, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38818568

RESUMO

The Finkel-Biskis-Jinkins Osteosarcoma (c-Fos; encoded by FOS) plays an important role in several cardiovascular diseases, including atherosclerosis and stroke. However, the relationship between FOS and venous thromboembolism (VTE) remains unknown. We identified differentially expressed genes in Gene Expression Omnibus dataset, GSE48000, comprising VTE patients and healthy individuals, and analysed them using CIBERSORT and weighted co-expression network analysis (WGCNA). FOS and CD46 expressions were significantly downregulated (FOS p = 2.26E-05, CD64 p = 8.83E-05) and strongly linked to neutrophil activity in VTE. We used GSE19151 and performed PCR to confirm that FOS and CD46 had diagnostic potential for VTE; however, only FOS showed differential expression by PCR and ELISA in whole blood samples. Moreover, we found that hsa-miR-144 which regulates FOS expression was significantly upregulated in VTE. Furthermore, FOS expression was significantly downregulated in neutrophils of VTE patients (p = 0.03). RNA sequencing performed on whole blood samples of VTE patients showed that FOS exerted its effects in VTE via the leptin-mediated adipokine signalling pathway. Our results suggest that FOS and related genes or proteins can outperform traditional clinical markers and may be used as diagnostic biomarkers for VTE.


Assuntos
Biologia Computacional , MicroRNAs , Neutrófilos , Proteínas Proto-Oncogênicas c-fos , Tromboembolia Venosa , Humanos , MicroRNAs/genética , MicroRNAs/sangue , MicroRNAs/metabolismo , Neutrófilos/metabolismo , Tromboembolia Venosa/genética , Tromboembolia Venosa/metabolismo , Tromboembolia Venosa/sangue , Biologia Computacional/métodos , Proteínas Proto-Oncogênicas c-fos/genética , Proteínas Proto-Oncogênicas c-fos/metabolismo , Regulação da Expressão Gênica , Masculino , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Feminino , Biomarcadores/sangue , Biomarcadores/metabolismo
18.
Cancer Med ; 13(11): e7309, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38819439

RESUMO

INTRODUCTION: Identification of molecular biomarkers in the saliva and serum of oral cavity cancer patients represents a first step in the development of essential and efficient clinical tools for early detection and post-treatment monitoring. We hypothesized that molecular analyses of paired saliva and serum samples from an individual would likely yield better results than analyses of either serum or saliva alone. MATERIALS AND METHODS: We performed whole-transcriptome and small non-coding RNA sequencing analyses on 32 samples of saliva and serum collected from the same patients with oral squamous cell carcinoma (OSCC) and healthy controls (HC). RESULTS: We identified 12 novel saliva and serum miRNAs and a panel of unique miRNA and mRNA signatures, significantly differentially expressed in OSCC patients relative to HC (log2 fold change: 2.6-26.8; DE: 0.02-0.000001). We utilized a combined panel of the 10 top-deregulated miRNAs and mRNAs and evaluated their putative diagnostic potential (>87% sensitivity; 100% specificity), recommending seven of them for further validation. We also identified unique saliva and serum miRNAs associated with OSCC and smoking history (OSCC smokers vs. never-smokers or HC: log2 fold change: 22-23; DE: 0.00003-0.000000001). Functional and pathway analyses indicated interactions between the discovered OSCC-related non-invasive miRNAs and mRNAs and their targets, through PI3K/AKT/mTOR signaling. CONCLUSION: Our data support our hypothesis that using paired saliva and serum from the same individuals and deep sequencing analyses can provide unique combined mRNA and miRNA signatures associated with canonical pathways that may have a diagnostic advantage relative to saliva or serum alone and may be useful for clinical testing. We believe this data will contribute to effective preventive care by post-treatment monitoring of patients, as well as suggesting potential targets for therapeutic approaches.


Assuntos
Biomarcadores Tumorais , MicroRNAs , Neoplasias Bucais , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Saliva , Transdução de Sinais , Serina-Treonina Quinases TOR , Humanos , Neoplasias Bucais/genética , Neoplasias Bucais/sangue , Neoplasias Bucais/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Serina-Treonina Quinases TOR/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Feminino , Masculino , Biomarcadores Tumorais/genética , Saliva/metabolismo , Saliva/química , Fosfatidilinositol 3-Quinases/metabolismo , Fosfatidilinositol 3-Quinases/genética , Pessoa de Meia-Idade , MicroRNAs/genética , MicroRNAs/sangue , Transcriptoma , Regulação Neoplásica da Expressão Gênica , Perfilação da Expressão Gênica , Idoso , Pequeno RNA não Traduzido/genética , Pequeno RNA não Traduzido/sangue , Adulto , Estudos de Casos e Controles , Análise de Sequência de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/sangue , Carcinoma de Células Escamosas/metabolismo
19.
PLoS One ; 19(5): e0298536, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38820252

RESUMO

BACKGROUND: The early detection of breast cancer (BC) is receiving global attention, creating an urgent need for more sensitive and comprehensive strategies for preventive intervention, therapy assessment, and prognosis prediction. Aberrant expression of miRNAs has been observed in various malignancies and may be potential targets for therapy. Our study aims to examine the expression profiles of miR-375, miR-574-3p, and miR-122 in the sera of Egyptian women with BC, benign breast lesions, and a control group. We hope to determine if these miRNAs can serve as minimally invasive biomarkers for BC. METHODS: This is a case-control study in which 77 patients with newly diagnosed BC, 20 patients with benign breast tumors, and 30 normal healthy subjects as controls were recruited from the outpatient clinic of the National Cancer Institute. The assessment of miRNAs was conducted using RT-PCR (Applied Biosystems). RESULTS: The expression level of miRNA-122 was significantly upregulated in the BC group, while the expression levels of miRNA-574 and miRNA-375 showed significant downregulation in BC patients. Serum miR-122 and miRNA-375 were able to distinguish breast cancer from the benign and control groups in ROC curve analysis, with AUCs of 0.786 and 0.796, respectively. Our results also showed that serum miR-122 and miR-574 are significant predictor variables in the multivariate analysis, after adjusting for age. CONCLUSIONS: Our findings suggest that miR-122 may act as an onco-microRNA, while miR-574 and miR-375 may have a main tumour suppressor role. The studied miRNAs may serve as minimally invasive biomarkers for cases of breast cancer and as promising potential therapeutic targets for breast cancer.


Assuntos
Biomarcadores Tumorais , Neoplasias da Mama , MicroRNAs , Humanos , MicroRNAs/genética , MicroRNAs/sangue , Neoplasias da Mama/genética , Neoplasias da Mama/sangue , Feminino , Egito/epidemiologia , Pessoa de Meia-Idade , Adulto , Estudos de Casos e Controles , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/sangue , Regulação Neoplásica da Expressão Gênica , Curva ROC , Idoso
20.
PLoS One ; 19(5): e0303035, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38820355

RESUMO

Fracture non-unions affect many patients worldwide, however, known risk factors alone do not predict individual risk. The identification of novel biomarkers is crucial for early diagnosis and timely patient treatment. This study focused on the identification of microRNA (miRNA) related to the process of fracture healing. Serum of fracture patients and healthy volunteers was screened by RNA sequencing to identify differentially expressed miRNA at various times after injury. The results were correlated to miRNA in the conditioned medium of human bone marrow mesenchymal stromal cells (BMSCs) during in vitro osteogenic differentiation. hsa-miR-1246, hsa-miR-335-5p, and miR-193a-5p were identified both in vitro and in fracture patients and their functional role in direct BMSC osteogenic differentiation was assessed. The results showed no influence of the downregulation of the three miRNAs during in vitro osteogenesis. However, miR-1246 may be involved in cell proliferation and recruitment of progenitor cells. Further studies should be performed to assess the role of these miRNA in other processes relevant to fracture healing.


Assuntos
Biomarcadores , Diferenciação Celular , MicroRNA Circulante , Células-Tronco Mesenquimais , MicroRNAs , Osteogênese , Humanos , Osteogênese/genética , MicroRNAs/sangue , MicroRNAs/genética , Células-Tronco Mesenquimais/metabolismo , Biomarcadores/sangue , Masculino , MicroRNA Circulante/sangue , MicroRNA Circulante/genética , Feminino , Consolidação da Fratura/genética , Adulto , Fraturas Ósseas/sangue , Fraturas Ósseas/genética , Pessoa de Meia-Idade , Células Cultivadas , Proliferação de Células
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