Project description:Total RNA from 59 human plasma samples was used to generate signatures of extracellular vesicles used as basis for evaluation of potential disease states
Project description:Transcriptome profiling has emerged as a powerful tool to investigate the biomarker potential of human biofluids. However, despite an enormous interest in extracellular nucleic acids, total RNA sequencing methods to quantify RNA content outside cells are rare. Here, we evaluate the performance of the SMARTer Stranded Total RNA-Seq method in human platelet-rich plasma, platelet-poor plasma, urine, conditioned medium, and extracellular vesicles from these biofluids.
Project description:We have analyzed six different extracelular vesicles isolation protocols from plasma of patients coinfected with human immunodeficiency virus and hepatitis C virus (HIV/HCV). In addition, two different RNA isolation kit were used for each protocol, one phenol-based and another pehenol-free kit. Same library kit was used for all samples. Small RNA, including miRNAs, piwiRNA, and sncRNAs were detected.
Project description:Background: There is some evidence demonstrating the effect of psychological interventions in improvements in health biological parameters. To best of our knowledge, no study had addressed the impact of any psychological intervention on extracellular vesicles. In addition, Mindfulness-Based Cognitive Therapy (MBCT) and Emotion Focused Therapy for Cancer Recovery (EFT-CR) in the group have never been explored regarding extracellular vesicles and the effectiveness of these was not compared yet.
Objectives:
1. To explore and compare the effect of MBCT and EFT-CR on biological parameters and psychological variables in distressed people who have had breast, prostate and colorectal cancer;
2. In addition, we will explore the acceptability through recruitment and retention rates of MBCT and EFT-CR in group and evaluate whether these interventions are appropriate for a larger clinical trial.
Methods: The design of this study is a parallel randomized controlled trial. Participants will be randomized into MBCT, EFT-CR or usual care. Outcome measures will be assessed before, at the end of the intervention (8 weeks) and follow-ups (24 and 52 weeks from the baseline moment).
Hypotheses: The researchers expected that both interventions will have an effect on extracellular vesicles and other study biomarkers as well as improvements in psychological outcomes, compared to treatment as usual (TAU) group. Regarding the comparative effectiveness, we did not have evidence to hypothesize which one of the interventions will be superior in both biological (extracellular vesicles) and psychological outcomes.
Contribution for practice: The results of this preliminary study would permit to know if there are benefits of these psychological interventions on changes in extracellular vesicles and on psychological outcomes related to health. In addition, this study will permit to determine the acceptability of conducting a larger randomized controlled trial.
Project description:RNA profiling has emerged as a powerful tool to investigate the biomarker potential of human biofluids. However, despite enormous interest in extracellular nucleic acids, RNA sequencing methods to quantify the total RNA content outside cells are rare. Here, we evaluate the performance of the SMARTer Stranded Total RNA-Seq method in human platelet-rich plasma, platelet-free plasma, urine, conditioned medium, and extracellular vesicles (EVs) from these biofluids. We found the method to be accurate, precise, compatible with low-input volumes and able to quantify a few thousand genes. We picked up distinct classes of RNA molecules, including mRNA, lncRNA, circRNA, miscRNA and pseudogenes. Notably, the read distribution and gene content drastically differ among biofluids. In conclusion, we are the first to show that the SMARTer method can be used for unbiased unraveling of the complete transcriptome of a wide range of biofluids and their extracellular vesicles.
Project description:Previous studies have shown that the circulating miRNA signatures are altered in plasma-derived extracellular vesicles of individuals with type 1 diabetes. These alterations in the miRNA profile could serve as a potential biomarker applicable in clinical practice for monitoring disease status in lactating mothers with type 1 diabetes during the postpartum period. In the present study, we investigate the profiles of extracellular vesicle-derived miRNAs in circulation in a cohort of lactating mothers with and without type 1 diabetes.
Project description:The seminal plasma contains large quantities of extracellular vesicles (EVs). However, the role of these EVs and their interactions with sperms are not clear. To identify the important molecules affecting sperm motility in EVs, we sequenced the EVs in the seminal plasma of Yorkshire boars with different sperm motility using whole RNA sequence.
Project description:Cell-free messenger RNA (cf-mRNA) has shown promising diagnostic potentials. Unlike well-characterized extracellular miRNA, how cf-mRNA is protected and its potential biological significance remains unknown. To characterize the carriers of cf-mRNA in human plasma and their association with cancers, we performed transcriptome analysis of cf-mRNA in size-fractionated plasma from patients with lung cancer, liver cancer and multiple myeloma, and healthy donors. We found that the majority of cf-mRNA was enriched in extracellular vesicles (EVs) and was protected in EVs with remarkable stability in RNase-rich environments. We observed specific enrichment patterns of cf-mRNA in each vesicular and non-vesicular subpopulation in cancers. These EV-enriched differentiating genes are associated with specific biological pathways, such as immune systems, liver function, and oxygen transport in lung cancer, liver cancer, and multiple myeloma, respectively. Our results suggest that dissecting the complexity of EVs subpopulations illuminates their biological significance and becomes a promising approach for novel biomarker development.