CoChat
Sign Up
Cited by 69Extracellular vesicles in diseaseNeuroinflammation and Neurodegeneration MechanismsNanoparticle-Based Drug Delivery

Engineered Extracellular Vesicles Loaded With miR-124 Attenuate Cocaine-Mediated Activation of Microglia

Open access
Chivero, Ernest T., Liao, Ke, Niu, Fang, Tripathi, Ashutosh, Tian, Changhai, Buch, Shilpa, Hu, Guoku·2020·Frontiers in Cell and Developmental Biology
DOI 10.3389/fcell.2020.00573PMC PMC7409518
Open this paper in CoChat — free accountOpen PDF

Abstract

MicroRNA-124 (miR-124), a brain-enriched microRNA, is known to regulate microglial quiescence. Psychostimulants such as cocaine have been shown to activate microglia by downregulating miR-124, leading, in turn, to neuroinflammation. We thus rationalized that restoring the levels of miR-124 could function as a potential therapeutic approach for cocaine-mediated neuroinflammation. Delivering miRNA based drugs in the brain that are effective and less invasive, however, remains a major challenge in the field. Herein we engineered extracellular vesicles (EVs) and loaded them with miR-124 for delivery in the brain. Approach involved co-transfection of mouse dendritic cells with Dicer siRNA and RVG-Lamp2b plasmid to deplete endogenous miRNAs and for targeting the CNS, respectively. Mouse primary microglia (mPm) were treated with purified engineered EVs loaded with either Cy5-miR-124 or Cy5-scrambled miRNA oligos in the presence or absence of cocaine followed by assessing EV uptake and microglial activation. In vivo studies involved pretreating mice intranasally with engineered EVs followed by cocaine injection (20 mg/kg, i.p.). mPm exposed to EV-miR-124 exhibited reduced expression of miR-124 targets - TLR4 & STAT3 as well as ERK-1/2 and Iba1. In cocaine administered mice, EV-Cy5-miR-124 delivered intranasally were detected in the CNS and significantly reduced the expression of inflammatory markers TLR4, MYD88, STAT3 and NF-kB p65 while also downregulating the microglial activation marker, Iba1. Collectively, these findings suggest that engineered EVs can deliver miR-124 into the CNS, thereby alleviating cocaine-mediated microglial activation. Manipulating EV miRNAs can thus be envisioned as an efficient means for delivery of RNA-based therapeutics to target organs.

AI snapshot

Written from the paper’s full text.

TL;DR

Engineered dendritic cell-derived extracellular vesicles loaded with miR-124 were taken up by microglia in vitro and reached the brain after intranasal delivery in cocaine-treated mice, where they attenuated cocaine-mediated microglial activation. The findings support EV-based intranasal delivery of miR-124 as a potential approach for cocaine-associated neuroinflammation.

Key findings

  • In mouse primary microglia, EV-miR-124 reduced cocaine (10 μM)-induced upregulation of TLR4, STAT3, ERK1/2, and Iba1 compared with scrambled miRNA controls.
  • In C57BL/6N mice given cocaine (20 mg/kg, i.p. for 7 consecutive days), intranasal EV-Cy5-miR-124 (20 μg/100 μL) was detected in the CNS and reduced cocaine-induced increases in TLR4, MYD88, STAT3, NF-kB p65, and Iba1 in cortex/striatum (n = 4/group).
  • Cocaine reduced cortical miR-124 levels; engineered EVs ranged from 40 to 150 nm, expressed CD63 and Lamp2, and Dicer knockdown reduced EV miR-138 and miR-223 without significantly affecting EV production.

Methods

  • DC2.4 mouse dendritic cells were co-transfected with Dicer siRNA and RVG-Lamp2b plasmid; EVs were isolated by differential ultracentrifugation, characterized by nanoparticle tracking analysis and western blot, and transfected with Cy5-miR-124 or Cy5-scrambled oligos.
  • Mouse primary microglia were exposed to EVs with or without cocaine (10 μM); C57BL/6N male mice received cocaine or saline (20 mg/kg, i.p.) for 7 days and intranasal EVs on days 5–7, followed by brain tissue analysis via western blot, Iba1 immunofluorescence, and IVIS biodistribution.
  • Statistical comparisons used Student’s t-test and one-way ANOVA, with p < 0.05 considered significant.

Limitations

  • The authors state that EVs also contain proteins and lipids beyond miRNAs that could lead to off-target effects and need further characterization; they also note that other brain cell types take up EVs and that side effects of miR-124 in those cells require further study.
  • Evident from the text: in vivo experiments used n = 4/group and only male C57BL/6N mice, and the study reports molecular/microglial markers rather than behavioral or functional neuroinflammatory outcomes.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.3389/fcell.2020.00573

Cites

55 of 55 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 50 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    NF-kB p65 was downregulated in mice administered cocaine and EV-Cy5-miR-124 pre-treatment compared with mice administered cocaine or saline and control EVs.

    “The results demonstrated that NF-kB p65 and STAT3 were downregulated in mice administrated with cocaine and EV-Cy5-miR-124 pre-treatment compared with mice administered with cocaine or saline and control EVs, respectively (Figures 4C,D).”
    • Also stated in: § Discussion · plain
      “Increased levels of miR-124 were found to attenuate cocaine-mediated upregulation of TLR4, MYD88, NF-kB p65 and STAT3 – mediators critical for cocaine-mediated neuroinflammation.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: miR-124 regulates NF-kB p65 and STAT3.
  • Findingown data

    EV lysates contained exosomal markers CD63 and Lamp2b.

    “Immunoblotting of the EV lysates revealed the presence of exosomal markers CD63 and Lamp2b (Figure 1F).”
    Quote in the paperSubstantial claim
  • Findingown data

    In cocaine-administered mice, EV-Cy5-miR-124 pretreatment prevented cocaine-induced upregulation of Iba1 in the cortex and striatum.

    “As shown in Figure 3C, there was downregulation of Iba1 in the cortex of mice pretreated with EV-Cy5-miR-124 in the presence of cocaine compared with levels of Iba1 in control mice administered cocaine and control EVs.”
    • Also stated in: § Intranasal Delivery of Dendritic Cell-Derived EVs Loaded With Cy5-miR-124 Inhibits Cocaine-Mediated Activation of Microglia in vivo · plain
      “In mice pretreated with EVs-Cy5-miR-124 intranasally, however, cocaine failed to upregulate the expression of Iba1 (Figure 3D).”
    • Also stated in: § Intranasal Delivery of Dendritic Cell-Derived EVs Loaded With Cy5-miR-124 Inhibits Cocaine-Mediated Activation of Microglia in vivo · plain
      “We also analyzed striatal tissue sections and found similar findings as that shown for cortex, specifically that Cy5-miR-124 EVs abrogated cocaine-induced upregulation of Iba1 as evidenced in Figure 3F and quantified in Figure”
    Quote in the paperSubstantial claim
  • Findingown data

    Mouse primary microglia exposed to Cy5-miR-124 EVs for 1 h showed cytoplasmic and perinuclear Cy5 fluorescence, indicating EV uptake.

    “As shown in Figure 2A, mouse primary microglia exposed to EVs transfected with Cy5-miR-124 (1 h) demonstrated a cytoplasmic and perinuclear Cy5 fluorescence suggesting thereby that EVs were taken up by microglial cells (Figure 2A).”
    Quote in the paperSubstantial claim
  • Findingown data

    Cy5-miR-124 EVs abrogated cocaine-induced upregulation of Iba1 in the cortex.

    “shown for cortex, specifically that Cy5-miR-124 EVs abrogated cocaine-induced upregulation of Iba1 as evidenced in Figure 3F and quantified in Figure 3G.”
    • Also stated in: § Discussion · plain
      “Additionally, there was also reduced expression of the microglial marker Iba1 in the presence of EV-miR-124.”
    Quote in the paperSubstantial claim
  • Classified as probably not a substantial claim. 18
  • May combine two claims. 19

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    Normalizing the surface area of Iba1+ cells to the total number of Iba1+ cells yielded similar results.

    “In addition, we also normalized the surface area of Iba1+ cells to the total number of Iba1+ cells and obtained similar results as shown in Supplementary Figures 3C,D.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Co-transfection of mouse dendritic cells with Dicer siRNA and RVG-Lamp2b plasmid efficiently knocked down dicer and over-expressed Lamp2b.

    “As shown in Figures 1A,B, there was efficient knockdown of dicer and over-expression of Lamp2b in these cells following dicer siRNA and RVG-Lamp2b co-transfection.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Dendritic-cell derived EVs were taken up by mouse primary microglia in vitro and into the brains of mice following intranasal administration.

    “Dendritic-cell derived EVs were taken up by mouse primary microglia in vitro and as well as into the brains of mice following intranasal administration.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    miR-124 is a brain-enriched microRNA that is highly expressed in microglia and is critical for maintaining microglia in a quiescent state.

    “MicroRNA-124 (miR-124), a brain-enriched microRNA, is known to regulate microglial quiescence.”
    • Also stated in: § Introduction · plain
      “Additionally, miR-124 is highly expressed in microglia and is critical for maintaining microglia in a quiescent state (Ponomarev et al., 2011; Veremeyko et al., 2013; Svahn et al., 2016).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites Ponomarev et al., 2011 source

      Not checked in the free run.

    • cites Veremeyko et al., 2013 source
    • cites Svahn et al., 2016 source

  • May combine two claims.

    Premisecitation

    Cocaine activates microglia by downregulating miR-124 and upregulating its targets, leading to neuroinflammation.

    “Psychostimulants such as cocaine have been shown to activate microglia by downregulating miR-124, leading, in turn, to neuroinflammation.”
    • Also stated in: § Introduction · plain
      “In particular, cocaine has been shown to activate microglia both in in vitro and in vivo model systems by mechanisms involving dysregulation of microRNAs (miRNAs) and their target genes (Guo et al., 2016; Periyasamy et al., 2017).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Guo et al., 2016 source
    • cites Periyasamy et al., 2017 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Altered expression of miR-124 is associated with multiple neuroinflammatory disorders such as Parkinson's, Alzheimer's, and Huntington's disease.

    “Notably, altered expression of miR-124 has been associated with multiple neuroinflammatory disorders such as Parkinson’s (Kanagaraj et al., 2014; Wang et al., 2016; Dong et al., 2018; Yao et al., 2018, 2019), Alzheimer’s Diseases (Lukiw, 2007; Smith et al., 2011; Fang et al., 2012; Zhou et al., 2018), and Huntington’s disease (Johnson and Buckley, 2009).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Kanagaraj et al., 2014 source
    • cites Wang et al., 2016 source
    • cites Dong et al., 2018 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    EVs have been successfully utilized for in vivo delivery of siRNAs to specific cell types and tissues in mice.

    “EVs have also been successfully utilized for in vivo delivery of siRNAs to specific cell types and tissues in mice (Van Den Boorn et al., 2011; Hu et al., 2018; Liao et al., 2019; Zhao et al., 2020).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Van Den Boorn et al., 2011 source
    • cites Hu et al., 2018

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Liao et al., 2019 source

  • May combine two claims.

    Premisecitation

    Cocaine is associated with increased immune reactivity and inflammation in humans and neuroinflammation in animal models.

    “Cocaine is a highly potent and addictive brain stimulant that is associated with increased immune reactivity and inflammation in humans (Fox et al., 2012; Ersche et al., 2014; Levandowski et al., 2016; Moreira et al., 2016; Pianca et al., 2017) and neuroinflammation in animal models (Guo et al., 2015; Periyasamy et al., 2017).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites Fox et al., 2012 source
    • cites Ersche et al., 2014 source
    • cites Levandowski et al., 2016 source

  • May combine two claims.

    Premisecitation

    Exposure of microglia to cocaine downregulates miR-124 and upregulates TLR4, MYD88, and STAT3.

    “A recent study has shown that exposure of microglia to cocaine downregulated the expression of miR-124 with concomitant upregulation of its targets such as TLR4, MYD88 and STAT3 (Periyasamy et al., 2017).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Periyasamy et al., 2017 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Dendritic cells communicate with neighboring cells through soluble mediators, cell-to-cell-contact and vesicle exchange.

    “Dendritic cells are sentinel antigen-presenting cells of the immune system that are known to communicate with neighboring cells through soluble mediators, cell-to-cell-contact and vesicle exchange (Montecalvo et al., 2012) and via release of a vast number of EVs.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Montecalvo et al., 2012 source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Dendritic cells and their EVs possess MHC-I and MHC-II molecules as well as costimulatory molecules, which interact with immune cells.

    “In addition, dendritic cells and the EVs they produce possess MHC-I and MHC-II molecules as well as costimulatory molecules, which interact with immune cells (Pitt et al., 2016).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Pitt et al., 2016 source

  • May combine two claims.

    Study design and conductcitation

    Purity of isolated microglia was confirmed by immunohistochemical staining for Iba1 and was routinely >95%.

    “The purity of isolated microglia was confirmed by immunohistochemical staining for Iba1 and was routinely found to be >95% pure (Guo et al., 2015; Chivero et al., 2017).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Guo et al., 2015 source
    • cites Chivero et al., 2017 source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Dendritic cells and dendritic cell-derived EVs possess MHC-I and MHC-II molecules and costimulatory molecules, facilitating interaction with immune cells.

    “Both dendritic cells as well as EVs derived from dendritic cells are known to possess MHC-I and MHC-II molecules as well as costimulatory molecules, which, in turn, facilitates interaction with immune cells (Pitt et al., 2016).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Pitt et al., 2016 source

  • Classified as probably not a substantial claim.

    Premisecitation

    There is a close link between drug abuse and neuroinflammation.

    “Accumulating evidence suggests a close link between drug abuse and neuroinflammation (Crews et al., 2011; Clark et al., 2013).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Crews et al., 2011 source

      Not checked in the free run.

    • cites Clark et al., 2013 source

  • May combine two claims.

    Interpretationown data

    Engineered EVs can deliver miR-124 into the CNS, thereby alleviating cocaine-mediated microglial activation.

    “Collectively, these findings suggest that engineered EVs can deliver miR-124 into the CNS, thereby alleviating cocaine-mediated microglial activation.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    EVs can deliver miRNAs for treatment of various diseases.

    “Several studies have reported and reviewed the potential of EVs in the delivery of miRNAs for the treatment of various diseases (Zhuang et al., 2011; Vader et al., 2016; Rufino-Ramos et al., 2017; Osorio-Querejeta et al., 2018, 2020; Long et al., 2020).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites Zhuang et al., 2011 source
    • cites Vader et al., 2016 source
    • cites Rufino-Ramos et al., 2017 source

  • Classified as probably not a substantial claim.

    Premisecitation

    Finding an effective and less invasive method for delivery of microRNA-based drugs into the brain remains a major challenge.

    “Finding an effective and less invasive method for the delivery of microRNA-based drugs into the brain remains a major challenge in the field (Zhuang et al., 2011).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Zhuang et al., 2011 source

  • May combine two claims.

    Interpretationown data

    Dendritic cell-derived EVs can deliver miR-124 to the CNS, thereby alleviating cocaine-mediated microglial activation.

    “Our results thus suggest that dendritic cell-derived EVs can deliver miR-124 to the CNS, thereby alleviating cocaine-mediated microglial activation.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Besides miRs, EVs are endowed with cargo containing proteins and lipids that could lead to off-target effects.

  • May combine two claims.

    Interpretationown data

    Microglia can take up these EVs both in vitro and in vivo.

    “Our findings suggest that microglia can take up these EVs both in vitro and in vivo.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Intravenously injected RVG EVs delivered GAPDH siRNA specifically to neurons, microglia and oligodendrocytes in the brain, resulting in tissue-specific gene knockdown.

    “It was reported that intravenously injected RVG EVs delivered GAPDH siRNA specifically to the neurons, microglia and oligodendrocytes in the brain, resulting in tissue-specific gene knockdown (Alvarez-Erviti et al., 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Alvarez-Erviti et al., 2011 source

  • Classified as probably not a substantial claim.

    Premisereasoning

    Dendritic cell-derived EVs can communicate with several cell types.

    “Since dendritic cell-derived EVs can communicate with several cell types, herein, we isolated EVs from dendritic cells and loaded them with Cy5-miR-124 for uptake by microglia both in vitro and in vivo.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    Alvarez-Erviti et al. successfully expressed Lamp2b fused to RVG peptide in dendritic cells, followed by loading EVs with exogenous siRNA of GAPDH.

    “As a means to increase the specific delivery of EVs into the CNS, we followed the approach reported by Alvarez-Erviti et al. (2011) wherein the authors were able to successfully express Lamp2b, an EV membrane protein, fused to an rabies virus glycoprotein (RVG) peptide in dendritic cells, followed by loading the EVs isolated from these cells with exogenous siRNA of GAPDH.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Alvarez-Erviti et al. (2011) source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Dendritic cells and their EVs possess surface adhesion membrane proteins such as integrins, ICAM-1, and MFG-E8 which facilitate effective targeting and docking to recipient cells.

    “These cells and their EVs also possess a variety of surface adhesion membrane proteins such as integrins, immunoglobulin family member ICAM-1, and milk fat globule EGF factor 8 (MFG-E8) which facilitates effective targeting and docking to recipient cells (Pitt et al., 2016).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Pitt et al., 2016 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    The animal study was reviewed and approved by the UNMC Institutional Animal Care and Use Committee.

    “The animal study was reviewed and approved by the UNMC Institutional Animal Care and Use Committee.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    RVG peptide targets the alpha-7-subunit of the nicotinic acetylcholine receptor and facilitates transcytosis of EVs across the blood-brain barrier, with specific targeting of neural cells in the brain.

    “RVG peptide is known to target the alpha-7-subunit of the nicotinic acetylcholine receptor (Kim et al., 2010), and facilitates the transcytosis of the EVs across the blood-brain barrier, with specific targeting of neural cells in the brain (Loddo et al., 1966; Alvarez-Erviti et al., 2011).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites Kim et al., 2010 source
    • cites Loddo et al., 1966
    • cites Alvarez-Erviti et al., 2011 source

  • Classified as probably not a substantial claim.

    Study design and conductcitation

    In the current study, we attempted to limit off-target effects of miRNAs in EVs by knocking down dicer using siRNA approach.

    “In the current study, we attempted to limit the off-target effects of miRNAs in EVs by knocking down dicer using the siRNA approach.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    DC2.4 dendritic cell line exhibits features of dendritic cells including expression of dendritic cell-specific markers, cell morphology, and ability to phagocytose and present exogenous antigens on MHC class I and class II molecules.

    “Here we used EVs derived from the DC2.4 dendritic cell line that has been well characterized and exhibits features of dendritic cells including the expression of dendritic cell-specific markers, cell morphology and has the ability to phagocytose and present exogenous antigens on both MHC class I and class II molecules (Shen et al., 1997).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Shen et al., 1997 source

  • May combine two claims.

    Premisecitation

    miR-124 is highly expressed in microglia and is critical for maintaining microglia in a quiescent state.

    “It has been well-recognized that miR-124 is highly expressed in microglia and is critical for maintaining microglia in a quiescent state (Ponomarev et al., 2011; Veremeyko et al., 2013; Svahn et al., 2016).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites Ponomarev et al., 2011 source

      Not checked in the free run.

    • cites Veremeyko et al., 2013 source
    • cites Svahn et al., 2016 source

  • May combine two claims.

    Premisecitation

    Cocaine downregulates expression of miR-124 in microglia with concomitant upregulation of its targets KLF4 and TLR4.

    “Cocaine has been shown to downregulate expression of miR-124 in microglia with concomitant upregulation of its targets – KLF4 and TLR4 (Guo et al., 2016; Periyasamy et al., 2017).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Guo et al., 2016 source
    • cites Periyasamy et al., 2017 source

  • May combine two claims.

    Interpretationown data

    Intranasal delivery of EVs loaded with miR-124 could reach the brain, leading to downregulated expression of TLR4 and other miR-124 target genes, ultimately resulting in amelioration of microglial activation in cocaine-administered mice.

    “Our in vivo findings demonstrated that intranasal delivery of EVs loaded with miR-124 could reach the brain, leading, in turn, to downregulated expression of TLR4 and other miR-124 target genes, ultimately resulting in amelioration of microglial activation in cocaine-administered mice.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Besides miRs, EVs are endowed with cargo containing proteins and lipids that could lead to off-target effects.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Endogenous miRNAs in EVs affect cellular functions of recipient cells via various mechanisms.

    “Endogenous miRNAs in EVs have been shown to affect the cellular functions of recipient cells via various mechanisms (Bayraktar et al., 2017).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Bayraktar et al., 2017 source

  • Classified as probably not a substantial claim.

    Premisecitation

    Overexpression of lentivirus miR-124 by stereotactic injection inhibits microglial activation in cocaine-administered mice.

    “Interestingly, overexpression of lentivirus miR-124 by stereotactic injection was shown to inhibit microglial activation in cocaine-administered mice (Periyasamy et al., 2017).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Periyasamy et al., 2017 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Presence and enrichment of miRNAs in EVs have been well-documented.

    “Presence and enrichment of miRNAs in EVs have been well-documented (Penfornis et al., 2016; Bayraktar et al., 2017).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Penfornis et al., 2016; Bayraktar et al., 2017

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

33 of 33

  • The paper relies on this without support.

    Delivering miRNA-based drugs effectively and less invasively to the brain remains a major challenge.

    “Delivering miRNA based drugs in the brain that are effective and less invasive, however, remains a major challenge in the field.”

  • The paper relies on this without support.

    Cy5-scrambled RNA does not recognize any sequences in mouse transcriptomes.

    “Cy5-scrambled RNA -5′cy5-gaucgaaccuagacuaguggu-cy5-3′, that does not recognize any sequences in mouse transcriptomes.”

  • The paper relies on this without support.

    Iba1 is a marker of microglial activation.

    “downregulating the microglial activation marker, Iba1.”

  • The paper relies on this without support.

    ERK1/2 is a marker of microglial activation.

    “In the presence of miR-124-EVs, however, cocaine failed to upregulate the expression of both these markers.”

  • The paper relies on this without support.

    miR-124 regulates NF-kB p65 and STAT3.

    “Since miR-124 is also known to regulate NF-kB p65 and STAT 3, we also analyzed the expression of these mediators in these mice.”

  • The paper relies on this without support.

    Besides miRs, EVs are endowed with cargo containing proteins and lipids that could lead to off-target effects.

    “We do acknowledge that besides miRs, EVs are also endowed with cargo containing proteins and lipids that could lead to off-target effects that need to be yet characterized.”
6 of 6

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Since many disorders share the same proteins or micro RNAs as markers, it is impossible to have selective detection of diseases.

    “Also, since many disorders share the same proteins or micro RNAs as markers, it is impossible to have selective detection of diseases [313–317].”

    From Extracellular vesicles as nanotheranostic platforms for targeted neurological disorder interventions (2024) · cites this paper as [317]

    The Introduction and Discussion describe miRNAs as therapeutic cargo in extracellular vesicles, not as diagnostic markers. Searches for disease-marker overlap and selective detection found no passage addressing whether shared proteins or miRNAs make selective disease detection impossible; the References list studies of miR-124 in several conditions but do not establish that claim. (LLM, conf=0.77)

    Experimental — accuracy not yet measured

Cites

Flagged first, then by citations

1 of the 56 references couldn’t be matched to a known paper.

Cited by

Sorted by citations

Related

Authors
Chivero, E. T., Liao, K., Niu, F., Tripathi, A., Tian, C., Buch, S., & Hu, G.
Affiliations
University of Nebraska Medical Center (US)
Corresponding author
  • Buch, Shilpa
  • Hu, Guoku
ORCID
Venue
Frontiers in Cell and Developmental Biology
Publisher
Frontiers Media SA
Published
2020-07-30
Volume
8
Pages
573-573
ISSN
2296-634X (ISSN-L)
DOI
10.3389/fcell.2020.00573
PMC
PMC7409518
OpenAlex
W3046291254
Semantic Scholar
f5cf60a1e5ee0792d3b051a9d0acdee7a5b6394b
Cited by
69
References
56
Citations by year
2026: 7 · 2025: 9 · 2024: 11 · 2023: 13 · 2022: 17 · 2021: 11 · 2020: 1
FWCI
3.05
Type
journal article
Language
en
Open access
Licence
Locations
  • Frontiers in Cell and Developmental Biology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DigitalCommons@UNMC (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
National Institutes of Health (DA042704, DA046831, DA043138, DA050545)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
Keywords
engineered extracellular vesicles, miR-124, cocaine, microglial activation, neuroinflammation, microRNA delivery, intranasal drug delivery, dendritic cell transfection, TLR4, STAT3, MyD88, NF-kB, Iba1, RNA therapeutics

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-11-06.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-11-06.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review