Plasma Exosomes Research Articles

Autologous exosome facilitates load and target delivery of bioactive peptides to repair spinal cord injury.

Spinal cord injury (SCI) causes motor, sensory and automatic impairment due to rarely axon regeneration. Developing effective treatment for SCI in the clinic is extremely challenging because of the restrictive axonal regenerative ability and disconnection of neural elements after injury, as well as the limited systemic drug delivery efficiency caused by blood spinal cord barrier. To develop an effective non-invasive treatment strategy for SCI in clinic, we generated an autologous plasma exosome (AP-EXO) based biological scaffold where AP-EXO was loaded with neuron targeting peptide (RVG) and growth-facilitating peptides (ILP and ISP). This scaffold can be targeted delivered to neurons in the injured area and elicit robust axon regrowth across the lesion core to the levels over 30-fold greater than naïve treatment, thus reestablish the intraspinal circuits and promote motor functional recovery after spinal cord injury in mice. More importantly, in ex vivo, human plasma exosomes (HP-EXO) loaded with combinatory peptides of RVG, ILP and ISP showed safety and no liver and kidney toxicity in the application to nude SCI mice. Combining the efficacy and safety, the AP-EXO-based personalized treatment confers functional recovery after SCI and showed immense promising in biomedical applications in treating SCI. It is helpful to expand the application of combinatory peptides and human plasma derived autologous exosomes in promoting regeneration and recovery upon SCI treatment.

PROGNOSTIC VALUE OF LIQUID BIOPSY IN CRC: A LITERATURE REVIEW

Relevance: Liquid biopsy is a modern, quite appropriate, and promising method for diagnosing malignant neoplasms for oncology. The method allows us to determine the level of freely circulating tumor cells – micrometastases, tumor DNA, microRNA, and exosomes in blood plasma- and detect various genetic changes. A literature review of current scientific publications on liquid biopsy techniques, indexed in Medline, PubMed, and Medscape, was carried out as part of the work.
 The study aimed to review is to assess the prognostic significance of liquid biopsy, to determine the place of the method in current recommendations, and its expediency from the point of view of the practice.
 Methods: The information search was conducted in the Medline, PubMed, and Medscape databases, with a search depth of 8 years. Data from randomized controlled trials, clinical trials, reviews, systematic reviews, and meta-analyses were analyzed. The review includes both full-fledged articles in the public domain and abstracts to obtain complete information on the problem.
 Results: Liquid biopsy surpasses tissue biopsy in simplicity and speed of research, easy repeatability, and minimal invasiveness, as well as the possibility of dynamic monitoring of progression – the overall clonal transformation of the tumor and the emergence of resistance to treatment.
 The disadvantages of this method are low sensitivity, difficulty in correctly interpreting biomarkers and determining their specificity, and high risk of false positive and false negative results due to dormant tumor cells.
 Conclusion: At present, the Liquid biopsy method is relevant and in demand, but it needs to be tested on a validated sample of the main population, and in order to achieve effective clinical use, important work needs to be done to standardize both preanalytical and analytical procedures and generalize them for all components of liquid biopsy.

PROGNOSTIC VALUE OF LIQUID BIOPSY IN CRC: A LITERATURE REVIEW

Relevance: Liquid biopsy is a modern, quite appropriate, and promising method for diagnosing malignant neoplasms for oncology. The method allows us to determine the level of freely circulating tumor cells – micrometastases, tumor DNA, microRNA, and exosomes in blood plasma- and detect various genetic changes. A literature review of current scientific publications on liquid biopsy techniques, indexed in Medline, PubMed, and Medscape, was carried out as part of the work.
 The study aimed to review is to assess the prognostic significance of liquid biopsy, to determine the place of the method in current recommendations, and its expediency from the point of view of the practice.
 Methods: The information search was conducted in the Medline, PubMed, and Medscape databases, with a search depth of 8 years. Data from randomized controlled trials, clinical trials, reviews, systematic reviews, and meta-analyses were analyzed. The review includes both full-fledged articles in the public domain and abstracts to obtain complete information on the problem.
 Results: Liquid biopsy surpasses tissue biopsy in simplicity and speed of research, easy repeatability, and minimal invasiveness, as well as the possibility of dynamic monitoring of progression – the overall clonal transformation of the tumor and the emergence of resistance to treatment.
 The disadvantages of this method are low sensitivity, difficulty in correctly interpreting biomarkers and determining their specificity, and high risk of false positive and false negative results due to dormant tumor cells.
 Conclusion: At present, the Liquid biopsy method is relevant and in demand, but it needs to be tested on a validated sample of the main population, and in order to achieve effective clinical use, important work needs to be done to standardize both preanalytical and analytical procedures and generalize them for all components of liquid biopsy.

CrebH protects against liver injury associated with colonic inflammation via modulation of exosomal miRNA

BackgroundHepatic liver disease, including primary sclerosing cholangitis (PSC), is a serious extraintestinal manifestations of colonic inflammation. Cyclic adenosine monophosphate (cAMP)-responsive element-binding protein H (CrebH) is a transcription factor expressed mostly in the liver and small intestine. However, CrebH’s roles in the gut–liver axis remain unknown.MethodsInflammatory bowel disease (IBD) and PSC disease models were established in wild-type and CrebH−/− mice treated with dextran sulfate sodium, dinitrobenzene sulfonic acid, and diethoxycarbonyl dihydrocollidine diet, respectively. RNA sequencing were conducted to investigate differential gene expression. Exosomes were isolated from plasma and culture media. miRNA expression profiling was performed using the NanoString nCounter Mouse miRNA Panel. Effects of miR-29a-3p on adhesion molecule expression were investigated in bEnd.3 brain endothelial cells.ResultsCrebH−/− mice exhibited accelerated liver injury without substantial differences in the gut after administration of dextran sulfate sodium (DSS), and had similar features to PSC, including enlarged bile ducts, enhanced inflammation, and aberrant MAdCAM-1 expression. Furthermore, RNA-sequencing analysis showed that differentially expressed genes in the liver of CrebH−/− mice after DSS overlapped significantly with genes changed in PSC-liver. Analysis of plasma exosome miRNA isolated from WT and CrebH−/− mice indicates that CrebH can contribute to the exosomal miRNA profile. We also identified miR-29a-3p as an effective mediator for MAdCAM-1 expression. Administration of plasma exosome from CrebH−/− mice led to prominent inflammatory signals in the liver of WT mice with inflammatory bowel disease (IBD).ConclusionsCrebH deficiency led to increased susceptibility to IBD-induced liver diseases via enhanced expression of adhesion molecules and concomitant infiltration of T lymphocytes. Exosomes can contribute to the progression of IBD-induced liver injury in CrebH−/− mice. These study provide novel insights into the role of CrebH in IBD-induced liver injury.

Long non-coding RNA and circular RNA and coding RNA profiling of plasma exosomes of osteosarcoma by RNA seq

Osteosarcoma (OS) is a primary bone tumor with high malignancy and the mechanism of hematogenous metastasis in OS is still not clear. The plasma exosomes derived from osteosarcoma play a key role in the process of tumor metastasis. Here, we established RNA-seq dataset for lncRNAs, circRNAs and mRNAs in plasma exosomes from 10 OS patients and 5 healthy donors. A total of 329.52 Gb of clean data was obtained. Besides, 1754 lincRNAs, 7096 known and 1935 new circRNA was identified. Finally, gene expression profiles and differentially expressed genes (DEGs) were analyzed among these 15 samples. There were 331 DEGs of mRNA, 132 of lincRNA and 489 of circRNA was obtained, respectively. This data set provides a significant resource for relevant researchers to excavate potential dysregulated lncRNAs, circRNAs and mRNAs of plasma exosomes in OS versus normal conditions.

Plasma exosomes and contained MiRNAs affect the reproductive phenotype in polycystic ovary syndrome.

Anovulation is the main feature of infertile women with polycystic ovary syndrome (PCOS), and there is very limited understanding of the role of plasma exosomes and miRNAs in it. To identify the effect of PCOS patients' plasma exosomes and exosomal miRNAs, we isolated plasma exosomes of PCOS patients and normal women and injected into 8-week-old ICR female mice via tail vein. The changes in estrus cycle, serum hormone levels, and ovarian morphology were observed. KGN cells were cultured and transfected with mimics and inhibitors of differentially expressed exosomal miRNAs (miR-18a-3p, miR-20b-5p, miR-106a-5p, miR-126-3p, and miR-146a-5p) and then tested for steroid hormone synthesis, proliferation, and apoptosis. The results showed that female ICR mice injected with plasma exosomes from PCOS patients presented ovarian oligo-cyclicity. Hormone synthesis and proliferation of granulosa cells were affected by differentially expressed PCOS plasma-derived exosomal miRNAs, of which miR-126-3p having the most evident effect. MiR-126-3p affected the proliferation of granulosa cells by inhibiting PDGFRβ and its downstream PI3K-AKT pathway. Our results demonstrated plasma exosomes and contained miRNAs in PCOS patients affect the estrus cycle of mice, hormone secretion, and proliferation of granulosa cells. This study provides a novel understanding about the function of plasma exosomes and exosomal miRNAs in PCOS.

Plasma exosomes reflect myocardial injury detected by cardiac magnetic resonance in STEMI patients

Abstract Funding Acknowledgements Type of funding sources: None. Background Cardiac magnetic resonance (CMR) is the gold standard technique to detect microvascular obstruction (MVO) and myocardial salvage index (MSI) in patients with ST-segment elevation myocardial infarction (STEMI) treated by primary percutaneous coronary intervention (pPCI). Exosomes are lipid vesicles released by all cells that reflect the metabolic status of the releasing cells, thus representing potential biomarkers of disease. The aim of this study is to evaluate the correlation between exosome profile and myocardial infarction characteristics as detected by CMR in patients after STEMI. Methods Exosomes were isolated from plasma of 42 patients by different commercial kits, their concentration and size distribution determined by Nanoparticle Tracking Analysis, exosomal surface epitopes by the MACSPlex Exosome Kit and GPIIb-IIIa expression by ELISA kit. Data obtained were correlated with parameters of myocardial damage (STEMI location, culprit lesion, time to revascularization, MVO and MSI). Results Levels of circulating plasma exosomes were higher in patients with anterior STEMI (p = 0.0002) and when the culprit lesion was located in LAD (p = 0.045). In addition, a high number of exosomes was measured in the subset of patients who underwent a late revascularization (> 3 h from onset of symptoms; p = 0.038). Patients with MVO and low MSI (< 0.5) have smaller exosome (p = 0.0015 and p = 0.014, respectively). By analysing the expression of 37 surface protein markers, the strongest signals are plateletspecific markers (CD41b and CD42a) followed by endothelial and immune cells markers. Interestingly, a lower expression of platelet marker GPIIb-IIIa was detected in both patients with MVO and anterior STEMI (p = 0.039 and p = 0.0038, respectively). Univariate analysis demonstrated that exosome size and GPIIb-IIIa were independent predictors of MVO/MSI and ROC analysis demonstrated that dimension and GPIIb-IIIa can identify patients with MVO/MSI (AUC 0.75 and 0.71, respectively). Conclusion The plasma exosome profile well reflects CMR-assessed myocardial injury after STEMI. In particular, the exosome dimension and the expression of platelet marker GPIIb-IIIa are independently associated with MVO. Future studies with larger populations are required to confirm the role of platelet-derived exosomesin risk stratification after STEMI.

A group of three miRNAs can act as candidate circulating biomarkers in liquid biopsies from melanoma patients.

Staging of melanoma and follow up after melanoma diagnosis aims at predicting risk and detecting progression or recurrence at early stage, respectively in order to timely start and/or change treatment. Tumor thickness according to Breslow, status of the sentinel node and value of the lactate dehydrogenase (LDH) are well-established prognostic markers for metastatic risk, but reliable biomarkers identifying early recurrence or candidates who may benefit best from medical treatment are still warranted. Liquid biopsy has emerged to be a suitable method for identifying biomarkers for early cancer diagnosis, prognosis, therapeutic response prediction, and patient follow-up. Liquid biopsy is a blood-based non-invasive procedure that allows analyzing circulating analytes, including extracellular vesicles. In this study we have explored the use of 7 miRNAs, namely hsa-miR-149-3p, hsa-miR-150-5p, hsa-miR-21-5p, hsa-miR-200c-3p, hsa-miR-134-5p, hsa-miR-144-3p and hsa-miR-221-3p in plasma exosomes to discriminate melanoma patients from controls without melanoma in a cohort of 92 individuals. Our results showed that three out seven miRNAs, namely hsa-miR-200c-3p, hsa-miR-144-3p and hsa-miR-221-3p were differentially expressed in plasma-derived exosomes from melanoma patients and controls. Furthermore, the expression of the three miRNAs may be a promising ancillary tool as a melanoma biomarker, even for discriminating between nevi and melanoma.

Characteristics of plasma exosomes in drug-resistant tuberculosis patients

BackgroundIncreasing prevalence of drug-resistant tuberculosis (DR-TB) poses a major challenge to the early detection and effective control of tuberculosis (TB). Exosomes carrying proteins and nucleic acid mediate intercellular communication between host and pathogen including Mycobacterium tuberculosis. However, molecular events of exosomes indicating the status and development of DR-TB remain unknown. This study determined the proteomics of exosome in DR-TB and explored the potential pathogenesis of DR-TB. MethodsPlasma samples were collected from 17 DR-TB patients and 33 non-drug-resistant tuberculosis (NDR-TB) patients using grouped case-control study design. After exosomes of plasma were isolated and confirmed by compositional and morphological measurement for exosomal characteristics, a label-free quantitative proteomics of exosomes was performed and differential protein components were determined via bioinformatics analysis. ResultsCompared with the NDR-TB group, we identified 16 up-regulated proteins and 10 down-regulated proteins in the DR-TB group. The down-regulated proteins were mainly apolipoproteins and mainly enriched in cholesterol metabolism-related pathways. Apolipoproteins family including APOA1, APOB, APOC1 were key proteins in protein-protein interaction network. ConclusionDifferentially expressed proteins in the exosomes may indicate the status of DR-TB from NDR-TB. Apolipoproteins family including APOA1, APOB, APOC1 may be involved in the pathogenesis of DR-TB by regulating cholesterol metabolism via exosomes.

CircEGFR reduces the sensitivity of pirarubicin and regulates the malignant progression of triple-negative breast cancer via the miR-1299/EGFR axis

Circular RNAs (circRNAs) have been found to be involved in cancer progression and chemotherapy sensitivity. However, the biological function of circRNAs in triple-negative breast cancer (TNBC) and its effect on the sensitivity to pirarubicin (THP) chemotherapy are still unclear. CircEGFR (hsa_circ_0080220) was screened and verified by bioinformatics analysis, proving it was highly expressed in TNBC cell lines, patient tissues, and plasma exosomes, and was associated with poor prognosis of patients. The expression level of circEGFR in patient tissue has potential diagnostic value to distinguish TNBC tissue from normal breast tissue. In vitro studies confirmed that overexpression of circEGFR promoted the proliferation, migration, invasion, and EMT of TNBC cells and decreased the sensitivity of THP treatment while silencing circEGFR showed the opposite effect. The circEGFR/miR-1299/EGFR pathway was cascaded and verified. CircEGFR regulated malignant progression of TNBC by regulating EGFR via sponging miR-1299. THP can inhibit the malignant phenotype of MDA-MB-231 cells by downregulating the expression of circEGFR. In vivo studies confirmed that overexpression of circEGFR can promote tumor growth and EMT and reduce tumor sensitivity to THP treatment. Silencing circEGFR inhibited the malignant progression of the tumor. These results revealed circEGFR is a promising biomarker for TNBC diagnosis, therapeutic and prognosis.

MicroRNA-23a-3p Is Upregulated in Plasma Exosomes of Bulbar-onset ALS Patients and Targets ERBB4

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease related to the progressive death of motor neurons. Understanding the pathogenesis of ALS continues to provide considerable challenges. Bulbar-onset ALS involves faster functional loss and shorter survival time than spinal cord-onset ALS. However, debate is ongoing regarding typical plasma miRNA changes in ALS patients with bulbar onset. Exosomal miRNAs have not yet been described as a tool for bulbar-onset ALS diagnosis or prognosis prediction. In this study, candidate exosomal miRNAs were identified by small RNA sequencing using samples from patients with bulbar-onset ALS and healthy controls. Potential pathogenic mechanisms were identified through enrichment analysis of target genes for differential miRNAs. Expression of miR-16-5p, miR-23a-3p, miR-22-3p, and miR-93-5p was significantly up-regulated in plasma exosomes from bulbar-onset ALS patients compared with healthy control subjects. Among them, miR-16-5p and miR-23a-3p were significantly lower in spinal-onset ALS patients than those with bulbar-onset. Furthermore, up-regulation of miR-23a-3p in motor neuron-like NSC-34 cells promoted apoptosis and inhibited cell viability. This miRNA was found to directly target ERBB4 and regulate the AKT/GSK3β pathway. Collectively, the above miRNAs and their targets are related to the development of bulbar-onset ALS. Our research indicates that miR-23a-3p might have an effect on motor neuron loss observed in bulbar-onset ALS and may be a novel target for the therapy of ALS in the future.

Plasma Exosome Analysis for Protein Mutation Identification Using a Combination of Raman Spectroscopy and Deep Learning.

Protein mutation detection using liquid biopsy can be simply performed periodically, making it easy to detect the occurrence of newly emerging mutations rapidly. However, it has low diagnostic accuracy since there are more normal proteins than mutated proteins in body fluids. To increase the diagnostic accuracy, we analyzed plasma exosomes using nanoplasmonic spectra and deep learning. Exosomes, a promising biomarker, are abundant in plasma and stably carry intact proteins originating from mother cells. However, the mutated exosomal proteins cannot be detected sensitively because of the subtle changes in their structure. Therefore, we obtained Raman spectra that provide molecular information about structural changes in mutated proteins. To extract the unique features of the protein from complex Raman spectra, we developed a deep-learning classification algorithm with two deep-learning models. Consequently, controls with wild-type proteins and patients with mutated proteins were classified with high accuracy. As a proof of concept, we discriminated the lung cancer patients with mutations in the epidermal growth factor receptor (EGFR), L858R, E19del, L858R + T790M, and E19del + T790M, from controls with an accuracy of 0.93. Moreover, the protein mutation status of the patients with primary (E19del, L858R) and secondary (+T790M) mutations was clearly monitored. Overall, our technique is expected to be applied as a novel method for companion diagnostic and treatment monitoring.

Effect of circulating exosomes in patients with sepsis on T cell function

To investigate the effect of circulating exosomes (EXO) on T cell function in patients with sepsis. Plasma EXO were obtained by ultracentrifugation from 10 patients with sepsis admitted to the emergency intensive care unit of Guangdong Provincial People's Hospital Affiliated to Southern Medical University. Transmission electron microscopy observation, nanoparticle tracking analysis (NTA), and Western blotting were used to detect EXO markers to identify their characteristics. Furthermore, peripheral blood mononuclear cells (PBMC) were isolated from the peripheral blood of 5 healthy volunteers, primary T cells were sorted by magnetic beads and expanded in vitro. After 24 hours of intervention with different doses (0, 1, 2.5, 5, 10 mg/L) of circulating EXO in patients with sepsis, T-cell activity was assessed using a cell counting kit-8 (CCK-8). The expression of T cell activation indicators CD69 and CD25 were observed using flow cytometry. Additional evaluations were performed on immunosuppressive indicators including the expression of programmed cell death 1 (PD-1) in CD4+ T cells and the proportion of regulatory T cell (Treg). The identification results confirmed that the successful isolation of EXO from the plasma of sepsis patients. The expression level of circulating EXO in sepsis patients was higher than that in healthy control group (mg/L: 48.78±5.14 vs. 22.18±2.25, P < 0.01). After 24 hours of intervention with 5 mg/L of plasma EXO from sepsis patients, T cells activity began to show suppression [(85.84±0.56)% vs. (100.00±0.00)%, P < 0.05]. As the dosage increased, after 24 hours of intervention with 10 mg/L of EXO, T cells activity was significantly suppressed [(72.44±2.36)% vs. (100.00±0.00)%, P < 0.01]. Compared with the healthy control group, after T cells intervention with plasma EXO from sepsis patients, the expression of early activation marker CD69 was significantly reduced [(52.87±1.29)% vs. (67.13±3.56)%, P < 0.05]. Meanwhile, there was an upregulation of PD-1 expression in T cells [(57.73±3.06)% vs. (32.07±0.22)%, P < 0.01] and an increase in the proportion of Treg [(54.67±1.19)% vs. (24.60±3.51)%, P < 0.01]. However, the expression of the late activation marker CD25 remained stable [(84.77±3.44)% vs. (85.93±2.32)%, P > 0.05]. Circulating EXO in sepsis patients induce T cell dysfunction, which may be a novel mechanism lead to immunosuppression in sepsis.

Expression and Clinical Significance of Exosome Derived MiR-181b-5p in Children with Acute Lymphoblastic Leukemia

To explore the expression level of exosome derived miR-181b-5p in different disease stages of children with acute lymphoblastic leukemia and its relationship with clinical characteristics. Bone marrow plasma samples of 86 children with ALL were collected. Exosomes were extracted by exosome extraction kit, and RNA in exosomes was extracted by TRIzol method. The levels of miR-181b-5p in the blood plasma exosomes of the patients in the newly diagnosed group, relapse group, remission group and control group were detected by qRT- PCR. The difference of miR-181b-5p expression level in each group was compared and analyzed, and the relationship between miR-181b-5p expression level and clinical characteristics was analyzed. The expression level of exosomal miR-181b-5p in the newly diagnosed group and the relapsed group was significantly lower than that in the remission group and the control group (P< 0.05). The expression level of exosomal miR-181b-5p in T-ALL children was higher than that in B-ALL children (P<0.05). The expression level of plasma exosomal miR-181b-5p in male children was higher than that in female children (P<0.01). Exosome derived miR-181b-5p changes dynamically in the course of ALL children, and can be used as a marker miRNA to monitor disease status. Exosomes can transmit information in the tumor microenvironment and serve as a potential carrier for biomolecular targeted therapy.

BIOFLUID SPECIFICITY OF LONG NON-CODING RNA PROFILE IN HYPERTENSION: RELEVANCE OF EXOSOMAL FRACTION

Objective: Non-coding RNA (ncRNA)-mediated targeting of various genes regulates the molecular mechanisms of the pathogenesis of hypertension (HTN). However, very few circulating long ncRNAs (lncRNAs) have been reported to be altered in essential HTN. The aim of this study was to identify simultaneously the lncRNA profiles in circulating plasma and packaged them into exosomes associated with UAE in HTN using deep sequencing technology. We also assessed the effect of the biofluid origin on lncRNA signature and regulated pathways. Design and method: In a cohort of hypertensive patients with (n = 22) or without urinary albumin excretion (UAE) (n = 26), we analyzed by next generation sequencing the lncRNA profile associated to UAE. Then, long Non-Coding RNA target predictions and molecular pathways analyses were performed through GO terms and KEGG pathways. Results: Plasma exosomes showed higher diversity and fold change of lncRNAs than plasma, and low transcript overlapping was found between the two biofluids. The majority of unique differentially expressed lncRNA in the exosome fraction were downregulated (71%) in albuminuric patients and 61% in plasma samples. In addition, in plasma samples only 40% of statistically significant transcripts had a log2 fod change higher or equal to 2 or lower or equal to -2, compared with 100% top lncRNAs in exosomal fraction. Enrichment analysis identified different biological pathways regulated in plasma or exosome fraction, which were implicated in fatty acid metabolism, extracellular matrix, and mechanisms of sorting ncRNAs into exosomes, while plasma pathways were implicated in genome reorganization, interference with RNA polymerase, and as scaffolds for assembling transcriptional regulators. Conclusions: Our study found a biofluid specific lncRNA profile associated with albuminuria, with higher diversity in exosomal fraction, which identifies several potential targets that may be utilized to study mechanisms of albuminuria and cardiovascular damage.

BMD‐001, a nanoparticle containing miR‐485‐3p antisense oligonucleotide, blocks Alzheimer's disease progression

AbstractBackgroundAlzheimer’s disease (AD) is a form of dementia characterized by progressive memory decline and cognitive dysfunction, which affects more than 44 million people worldwide. Currently, there is no effective therapy for AD despite its increasing global incidence; thus, effective treatment strategies for AD are urgently needed.MethodIn a previous report, we confirmed that expression of miR‐485‐3p was increased in the brain tissues, CSF, and plasma exosomes of AD patients than in healthy controls. In addition, the therapeutic potential of downregulation of miR‐485‐3p was also shown in 5XFAD AD model mice. Based on this target validation, we developed a nanoparticle containing antisense oligonucleotide (ASO) targeting miR‐485‐3p, named BMD‐001, which has a high CNS‐targeting ability to cross the blood‐brain barrier (BBB). BMD‐001 was injected into 10‐month‐old 5XFAD mice by intravenous injection, once weekly for four weeks. Behavioral tests were performed at 11 months and their brain pathology was examined after 8‐week‐washout at 13 months.ResultTo evaluate in vivo pharmacokinetics of BMD‐001, the plasma levels of Cy5‐labeled ASO encapsulated in the nanoparticle were measured after intravenous injection along with a naked ASO without formulation of nanoparticle. The half‐life of the ASO in the nanoparticle was 93 min while that of the naked ASO was only 43 sec. Furthermore, we also observed that the brain accumulation of the fluorescent ASO in the nanoparticle was 64‐fold higher than the naked ASO and was maintained up to 16 hours, as monitored by in vivo imaging. After BMD‐001 injection, miR‐485‐3p expression was decreased in the brain tissue. We found that BMD‐001 enhanced Aβ clearance via phagocytosis of Aβ in vitro and in vivo. Furthermore, BMD‐001 reduced Aβ plaques, tau pathology, inflammasome, neuroinflammation, and cognitive decline in the 5XFAD mice.ConclusionCollectively, our findings suggest that BMD‐001 is delivered to the brain through the efficient BBB penetration. Furthermore, BMD‐001 can be a promising candidate for treatment of AD pathology and cognitive decline, establishing a new paradigm in the AD field.

Comparative proteomic profiling of plasma exosomes in patients with lung cancer with brain metastasis and liver metastasis.

e21123 Background: Brain or liver metastases are the main causes of lung cancer-related death. Predicting liver or brain metastases before having evidence from imaging of the tumors is challenging, but important for early patient intervention. Accumulating evidence has suggested that circulating exosomes in the blood could mediate cancer metastasis by transferring specific proteins to the target cells. Methods: Using liquid chromatography-MS/MS, we analyzed the proteomic profiles of plasma exosomes derived from 42 metastatic lung cancer patients (including 26 solitary brain metastasis (BM) and 16 solitary liver metastasis (LM)), 25 local advanced (LA) lung cancer patients without metastasis, and 5 healthy controls (HC), and explored organ-specific proteomic biomarkers. Results: 120 and 143 differentially expressed exosomal proteins were found to be the dysregulated candidates in BM and LM of lung cancer (BM-DEEPs, LM-DEEPs), as compared to the LA lung cancer samples, respectively. The bioinformatics analyses indicated the homogeneity and heterogeneity in BM-DEEPs and LM-DEEPs. In terms of homogeneity, they were mainly involved in the protein activation cascade, collagen-containing extracellular matrix, and ECM-receptor interaction. In terms of heterogeneity, the BM-DEEPs were significantly enriched in S100 protein and calcium-dependent protein, while the LM-DEEPs were mainly enriched in integrin, lipoprotein, heat shock protein and proteoglycans. Moreover, our study also revealed the heterogeneity in plasma-derived exosome proteomics in patients with non-small cell lung carcinoma (NSCLC) and small cell lung carcinoma (SCLC), which partly explained the different metastatic characteristics of NSCLC and SCLC. Additionally, we found that MUC5B and SELL could be used as a reliable diagnostic marker of BM, while APOH, CD81 and CCT5 could help to diagnose LM in lung cancer patients. Conclusions: For the first time, we revealed the comprehensive and comparative proteomic profiles of plasma-derived exosomes from lung cancer patients with solitary brain and liver metastasis. We further proposed some potential biomarkers and pathological mechanisms, which could serve as excellent resource for further studies investigating lung cancer metastasis.

Plasma versican and plasma exosomal versican as potential diagnostic markers for non-small cell lung cancer

Background and aimsThis study aimed to investigate the expression of plasma versican and plasma exosomal versican in non-small cell lung cancer (NSCLC) and its correlation with clinicopathological features, and to evaluate its diagnostic performance in NSCLC and its predictive function for NSCLC incidence and metastasis risk.Materials and methodsThere were 110 instances of NSCLC, 42 cases of benign lung disease, and 55 healthy controls from September 2018 to October 2020 at Tongji Hospital Affiliated to Tongji University. Blood was collected and plasma was separated before surgery, and plasma exosomes were extracted by ExoQuick kit. Morphological and molecular phenotype identification of exosomes was performed by transmission electron microscopy, Nanosight particle tracking analysis, and western blotting. Plasma versican and plasma exosomal versican were detected in all subjects to assess their expression levels and diagnostic value in NSCLC. Clinicopathological data were collected to explore correlations between abnormal plasma versican and plasma exosomal versican expression and clinicopathological parameters. Receiver operating characteristic (ROC) curve was used to judge its diagnostic performance in NSCLC, and binary logistic regression analysis was used to predict the risk of NSCLC incidence and metastasis.ResultsPlasma versican and plasma exosomal versican expression in NSCLC patients was significantly upregulated and was significantly higher in T3 + T4 patients compared with T1 + T2 patients (P < 0.05); the levels of plasma versican and plasma exosomal versican were positively correlated with lymph node metastasis, distant metastases (e.g., brain, bone), and mutation(e.g., EGFR,ALK)in NSCLC patients (all P < 0.05). Furthermore, ROC curve analysis showed that plasma versican and plasma exosomal versican had higher AUC values than NSE, CYFRA21-1, and SCC, and better diagnostic performance in NSCLC patients. However, the AUC and diagnostic performances of plasma versican and plasma exosomal versican in advanced-stage NSCLC patients were not shown to be significantly better than CEA. The results of binary logistic regression analysis showed that high levels of plasma exosomal versican had higher predictive value for lung cancer incidence, while high levels of plasma versican had higher predictive value for lung cancer metastasis.ConclusionOur findings showed that plasma versican and plasma exosomal versican might be potential diagnostic markers for NSCLC. High plasma exosomal versican expression can be used as a predictor of NSCLC risk and high plasma versican expression can be used as a predictor of NSCLC metastasis risk.

Transcriptional profiling of exosomes derived from plasma of canine with mammary tumor by RNA-seq analysis

Canine mammary tumor (CMT) are the second most common tumor in dogs. Exosomes can act as biomarkers for the early diagnosis of tumors, and also be involved in the pathogenesis and metastasis mechanism of tumors. The expression profile of exosomal RNA revealed that there were a total of 5547 differentially expressed mRNAs, and 196 differentially expressed lncRNAs. GO and KEGG enrichment analysis found that the differentially expressed mRNAs and lncRNA target genes were associated with metabolic processes, DNA replication, cell proliferation, cell junction, and cell adhesion. In conclusion, this study revealed lncRNA and mRNA expression profiles in exosomes derived from plasma of CMT and further annotated their potential functions. The data obtained in this study will also provide valuable resources for understanding lncRNA information in plasma exosomes of dogs with CMT, and contribute to the study of early diagnostic markers and pathogenesis of CMT.

Differential expression profiles of plasma exosomal microRNAs in dilated cardiomyopathy with chronic heart failure.

As one of the most prevalent heritable cardiovascular diseases, dilated cardiomyopathy (DCM) induces cardiac insufficiency and dysfunction. Although genetic mutation has been identified one of the causes of DCM, the usage of genetic biomarkers such as RNAs for DCM early diagnosis is still being overlooked. In addition, the alternation of RNAs could reflect the progression of the diseases, as an indicator for the prognosis of patients. Therefore, it is beneficial to develop genetic based diagnostic tool for DCM. RNAs are often unstable within circulatory system, leading to the infeasibility for clinical application. Recently discovered exosomal miRNAs have the stability that is then need for diagnostic purpose. Hence, fully understanding of the exosomal miRNA within DCM patients is vital for clinical translation. In this study, we employed the next generation sequencing based on the plasma exosomal miRNAs to comprehensively characterize the miRNAs expression in plasma exosomes from DCM patients exhibiting chronic heart failure (CHF) compared to healthy individuals. A complex landscape of differential miRNAs and target genes in DCM with CHF patients were identified. More importantly, we discovered that 92 differentially expressed miRNAs in DCM patients undergoing CHF were correlated with several enriched pathways, including oxytocin signalling pathway, circadian entrainment, hippo signalling pathway-multiple species, ras signalling pathway and morphine addiction. This study reveals the miRNA expression profiles in plasma exosomes in DCM patients with CHF, and further reveal their potential roles in the pathogenesis of it, presenting a new direction for clinical diagnosis and management of DCM patients with CHF.