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Cited by 227Neurogenesis and neuroplasticity mechanismsExtracellular vesicles in diseaseNeuroinflammation and Neurodegeneration Mechanisms

Microglia secrete miR-146a-5p-containing exosomes to regulate neurogenesis in depression

Open access
Fan, Cuiqin, Li, Ye, Lan, Tian, Wang, Wenjing, Long, Yifei, Yu, Shu Yan·2022·Molecular Therapy
DOI 10.1016/j.ymthe.2021.11.006PMC PMC8899528
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Abstract

Enhancing neurogenesis within the hippocampal dentate gyrus (DG) is critical for maintaining brain development and function in many neurological diseases. However, the neural mechanisms underlying neurogenesis in depression remain unclear. Here, we show that microglia transfer a microglia-enriched microRNA, miR-146a-5p, via secreting exosomes to inhibit neurogenesis in depression. Overexpression of miR-146a-5p in hippocampal DG suppresses neurogenesis and spontaneous discharge of excitatory neurons by directly targeting Krüppel-like factor 4 (KLF4). Downregulation of miR-146a-5p expression ameliorates adult neurogenesis deficits in DG regions and depression-like behaviors in rats. Intriguingly, circular RNA ANKS1B acts as a miRNA sequester for miR-146a-5p to mediate post-transcriptional regulation of KLF4 expression. Collectively, these results indicate that miR-146a-5p can function as a critical factor regulating neurogenesis under conditions of pathological processes resulting from depression and suggest that microglial exosomes generate new crosstalk channels between glial cells and neurons.

AI snapshot

Written from the paper’s full text.

TL;DR

Microglia-derived exosomes carrying miR-146a-5p transfer to hippocampal dentate gyrus neurons, where miR-146a-5p targets KLF4 to suppress neurogenesis and contribute to depression-like behaviors in CUMS rats. circANKS1B acts as a miR-146a-5p sequester, and manipulating the miR-146a-5p/KLF4/CDKL5 pathway alters neurogenesis and depression-like phenotypes.

Key findings

  • CUMS rats showed reduced sucrose preference and increased forced-swim immobility (n = 23 per group, p < 0.01), fewer newly developed DG cells (n = 6 per group, p < 0.01), and decreased sEPSC amplitude/frequency in DG neurons (n = 18 neurons from 7 control rats; n = 16 neurons from 6 CUMS rats; p < 0.001).
  • Serum-derived exosomes from CUMS rats had a double-membrane structure, median diameter 91.28 nm, and exosomal markers CD63/CD81/Alix; miR-146a-5p was upregulated in CUMS exosomes and was most closely related to signaling pathways regulating pluripotency of stem cells.
  • miR-146a-5p directly repressed wild-type KLF4 3′ UTR luciferase reporter activity (n = 6 per group; p < 0.0001). DG overexpression of miR-146a-5p suppressed neurogenesis and sEPSC activity, decreased KLF4/CDKL5, and increased p-STAT3; miR-146a-5p knockdown in CUMS rats increased neurogenesis and reversed depression-like behaviors.
  • circANKS1B was decreased in CUMS DG, bound miR-146a-5p, and regulated KLF4/CDKL5; circANKS1B knockdown reduced neurogenesis, while circANKS1B overexpression in CUMS rats increased newly developed cells and ameliorated depression-like behaviors.

Methods

  • Male Wistar rats (120–140 g) underwent 5 weeks of chronic unpredictable mild stress (CUMS); behavioral testing used sucrose preference and forced swim tests, with stereotaxic AAV injections into hippocampal DG and whole-cell patch-clamp recordings of DG granule cell sEPSCs.
  • Exosomes were isolated from rat serum by ultracentrifugation and precipitation; in vitro work used BV-2 microglia, LPS, GW4869, PLX3397, and cultured neural stem cells, with miRNA/circRNA high-throughput sequencing, qPCR, western blot, immunofluorescence, flow cytometry, dual-luciferase assays, and exosome labeling.
  • Data are means ± SEM, analyzed with GraphPad Prism 8.0; Student’s t tests for two groups, one-way ANOVA with Tukey’s post hoc for >2 groups, normality testing, and p < 0.05 significance. RNA-seq data: GEO GSE185777.

Limitations

  • The paper states it lacks definitive proof of glia-to-neuron shuttling of bioactive miR-146a-5p via exosomes; whether decreased KLF4 in the depression model is mediated by microglial exosomal miR-146a-5p needs further confirmation.
  • The paper notes that LPS-induced in vitro inflammation may differ from chronic stress-induced inflammation in vivo, and whether LPS stimulation functionally regulates neurogenesis via the HPA axis remains unclear; other conserved miR-146a-5p targets such as IRAK1 and TRAF6 were not investigated.
  • Evident from the text: all in vivo experiments used male Wistar rats, and no human cohort data are reported despite references to major depressive disorder.

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

Integrity signals

Registry

Crossref · 10.1016/j.ymthe.2021.11.006

Cites

61 of 61 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

7 of 37 cited sources checked · 4 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    miRNAs are important participants in the pathogenesis of depression.

    “recent evidence has emerged indicating that miRNAs are important participants in the pathogenesis of depression.25”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [25] Xiao, 2012, Prioritizing cancer-related key miRNA-target interactions by integrative genomics, Nucleic Acids Res. source

      The Introduction links miRNAs to the pathogenesis of cancer, not depression. Searches for depression and related psychiatric terms found no passage in our copy addressing a role for miRNAs in depression. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
  • Findingown data

    The frequency of sEPSC is markedly decreased in DG neurons of hippocampal slices in depressed versus control rats.

    “the amplitude and frequency of spontaneous excitatory post-synaptic current (sEPSC) were markedly decreased in DG neurons of hippocampal slices in depressed versus control rats (Figures 1E–1G).”
    Quote in the paperNot a substantial claim
  • Findingown data

    Overexpression of miR-146a-5p in hippocampal DG reduces sEPSC amplitude in DG neurons.

    “as were sEPSC frequencies and amplitudes (Figures 4G–4I).”
    Quote in the paperNot a substantial claim
  • Findingown data

    miR-146a-5p in CSF showed increased levels of expression in CUMS rats compared with control.

    “the presence of miR-146a-5p in CSF showed increased levels of expression in CUMS rats as compared with that in the control group (Figure S4C).”
    Quote in the paperNot a substantial claim
  • Findingown data

    Various miRNAs were significantly dysregulated in exosomes from CUMS versus control rats.

    “A heatmap showed that various miRNAs were significantly dysregulated in samples from CUMS versus control rats (Figures 2D and S3; and Table S1).”
    Quote in the paperSubstantial claim
  • Classified as probably not a substantial claim. 24
  • May combine two claims. 27
  • The quote only partly states this claim. 1

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

    Premisecitation

    Various miRNA alterations are present in cerebrospinal fluid, serum, and the ventral prefrontal cortex in patients with major depression disorder.

    “various miRNA alterations are present in cerebrospinal fluid, serum, and the ventral prefrontal cortex in patients with major depression disorder (MDD).26”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [26] Lopez, 2017, MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes, Nat. Commun. source

      The Results sections on vPFC dysregulation and blood-sample composition report altered miRNAs in the ventrolateral prefrontal cortex of depressed individuals and in blood and cell-free plasma after antidepressant treatment. They do not establish alterations in cerebrospinal fluid or serum; plasma is not serum. The blood findings are also specifically associated with treatment response, rather than MDD generally. No page numbers are available in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

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

    Premisecitation

    The decrease in hippocampal volume is positively correlated with duration of depression.

    “an effect often positively correlated with the duration of depression.6, 7, 8, 9”
    Quote in the paperNot a substantial claim2 of 4 sources checked
    • cites [6] Kempton, 2011, Structural neuroimaging studies in major depressive disorder meta-analysis and comparison with bipolar disorder, Arch. Gen. Psychiatry source
    • cites [7] Boldrini, 2018, Human hippocampal neurogenesis persists throughout aging, Cell Stem Cell source

      The Discussion reports stable dentate gyrus volume across ages in subjects without neuropsychiatric disease, not a relationship between hippocampal volume and depression duration. Searches for “duration of depression hippocampal volume” and “depression hippocampal volume” found no passage addressing the claimed correlation. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [8] Cole, 2011, Hippocampal atrophy in first episode depression: a meta-analysis of magnetic resonance imaging studies, J. Affect Disord. source
    • cites [9] Boldrini, 2013, Hippocampal granule neuron number and dentate gyrus volume in antidepressant-treated and untreated major depression, Neuropsychopharmacology source

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

    Premisecitation

    KLF4 is expressed in neuronal stem cells and regulates neuronal differentiation and migration in the brain.

    “One eukaryotic transcription factor, which is expressed in neuronal stem cells (NSCs) and regulates neuronal differentiation and migration in the brain, is that of KLF4.28”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [28] Cui, 2017, KLF4 knockdown attenuates TBI-induced neuronal damage through p53 and JAK-STAT3 signaling, CNS Neurosci. Ther. source

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

    Premisecitation

    Microglial degeneration is observed in depressed patients.

    “microglial degeneration is also observed in depressed patients.33”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] Brites, 2015, Neuroinflammation and depression: microglia activation, extracellular microvesicles and microRNA dysregulation, Front Cell Neurosci. source

  • May combine two claims.

    Premisecitation

    CDKL5 is highly expressed in the brain and deficiencies in CDKL5 inhibit the formation of new granular neurons.

    “cyclin-dependent kinase-like 5 (CDKL5) is highly expressed in the brain and deficiencies in CDKL5 inhibit the formation of new granular neurons.31”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [31] Fuchs, 2014, Loss of CDKL5 impairs survival and dendritic growth of newborn neurons by altering AKT/GSK-3 beta signaling, Neurobiol. Dis. source

  • May combine two claims.

    Findingown data

    Exosomes isolated from rat serum possessed a double membrane structure and a median diameter of 91.28 nm.

    “These vesicles possessed a double membrane structure (Figure 2A) and a median diameter of 91.28 nm (Figure 2B), characteristics which were suggestive of exosomes.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S4 (based). The quote only partly states this claim.

    Findingnone given

    Similar effects (increased exosomal miR-146a-5p) are observed in patients with major depressive disorder.

    “Similar effects are observed in patients with MDD.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In CUMS rats, behavioral responses are accompanied by alterations in neurogenesis and reductions in neuronal activity within the hippocampal DG area.

    “these behavioral responses were accompanied with alterations in neurogenesis and reductions in neuronal activity within the hippocampal DG area.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    miR-146a-4p is enriched and significantly increased in the exosomes of CUMS rats.

    “it was also enriched and significantly increased in the exosomes of CUMS rats.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    miR-146a-5p is present in cultured microglia and exosomes thereof, more abundantly under stimulation of LPS.

    “PCR results confirmed that miR-146a-5p is present in cultured microglia and exosomes thereof, more abundantly under stimulation of LPS.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingcitation

    KLF4 silencing induces depression-like behaviors and deficiencies in the development of new neurons.

    “KLF4 silencing induces depression-like behaviors and, consistent with previous studies, induces deficiencies in the development of new neurons.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Overexpression of circANKS1B enhances DG neurogenesis and ameliorates depression-like behaviors in CUMS rats.

    “an overexpression of circANKS1B enhanced DG neurogenesis and ameliorated depression-like behaviors in CUMS rats.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Silencing circANKS1B significantly attenuates neurogenesis and induces depression-like symptoms in normal rats.

    “silencing circANKS1B significantly attenuated neurogenesis and induced depression-like symptoms in normal rats”
    Quote in the paperSubstantial claim

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

    Premisecitation

    The size and density of neurons and glial cells in the dorsolateral prefrontal cortex are decreased in depression.

    “the size and density of neurons and glial cells in the dorsolateral prefrontal cortex were also decreased in depression.10”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [10] Fossati, 2004, Relevance of neuroplasticity and cellular resilience in bipolar disorder and depression: evidence from neuroimaging and cognitive studies, Eur. Neuropsychopharm. source

  • Classified as probably not a substantial claim.

    Premisecitation

    Microglia can secrete exosomes carrying miRNA capable of exerting acute impact upon adult neuroplasticity.

    “it has been reported that microglia can secrete exosomes carrying miRNA that are capable of exerting an acute impact upon adult neuroplasticity.17,18”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [17] McNeill, 2012, MicroRNAs shape the neuronal landscape, Neuron source

      Not checked in the free run.

    • cites [18] Prada, 2018, Glia-to-neuron transfer of miRNAs via extracellular vesicles: a new mechanism underlying inflammation-induced synaptic alterations, Acta Neuropathol. source

      Not checked in the free run.

  • May combine two claims.

    Study design and conductown data

    High expressions of exosomal marker proteins CD63, CD81, and Alix were present in serum-derived vesicles, while poor levels of GM130 and albumin were detected.

    “high expressions of the exosomal marker proteins, CD63, CD81, and Alix, were present in these vesicles. Meanwhile, poor levels of GM130 and albumin, markers of Golgi apparatus and lipoproteins, respectively, were detected by western blots”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Microglia participate in stem cell proliferation and survival through release of trophic factors and anti-inflammatory cytokines.

    “A previous study has reported that microglia participate in stem cell proliferation and survival through their release of important trophic factors and anti-inflammatory cytokines.16”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Cope, 2019, Adult neurogenesis, glia, and the extracellular matrix, Cell Stem Cell source

      Not checked in the free run.

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

    Premisecitation

    Exosomes can transport across the blood-brain barrier and play a key role in cellular communication, neurogenesis, and synaptic plasticity in the central nervous system.

    “The capacity for transport of exosomes across the blood-brain barrier enables them to play a key role in cellular communication, neurogenesis, and synaptic plasticity in the central nervous system.13,14”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [13] Fruhbeis, 2012, Emerging roles of exosomes in neuron-glia communication, Front Physiol. source

      Not checked in the free run.

    • cites [14] Xu, 2017, Neurons secrete miR-132-containing exosomes to regulate brain vascular integrity, Cell Res. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    Microglia perform this function at least by secreting exosomes to promote inter-neuronal communication.

    “these microglia appear to perform this function at least by secreting exosomes to promote inter-neuronal communication.20”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [20] Budnik, 2016, Extracellular vesicles round off communication in the nervous system, Nat. Rev. Neurosci. source

      Checked before — its earlier verdict is held back until a new check runs.

  • Classified as probably not a substantial claim.

    Premisecitation

    Exosomes can carry specific microRNAs, DNA, mRNA, and proteins to recipient cells.

    “Exosomes, which are a type of vesicle with a double membrane structure, can carry a variety of molecules, such as specific microRNAs, DNA, mRNA, and proteins, to recipient cells.12”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [12] Saadatpour, 2016, Glioblastoma: exosome and microRNA as novel diagnosis biomarkers, Cancer Gene Ther. source

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

    Premisecitation

    miRNAs are abundantly expressed in the brain, and miRNA dysfunction contributes to neurological diseases.

    “miRNAs have been shown to be abundantly expressed in the brain, and miRNA dysfunction contributes to neurological diseases.22,23”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [22] Long, 2012, MicroRNA-153 physiologically inhibits expression of amyloid-beta precursor protein in cultured human fetal brain cells and is dysregulated in a subset of Alzheimer disease patients, J. Biol. Chem. source
    • cites [23] Long, 2014, MicroRNA-339-5p down-regulates protein expression of beta-site amyloid precursor protein-cleaving enzyme 1 (BACE1) in human primary brain cultures and is reduced in brain tissue specimens of Alzheimer disease subjects, J. Biol. Chem. source

  • Classified as probably not a substantial claim.

    Premisecitation

    Microglia can secrete exosomes.

    “A diverse array of cells, including microglia, can secrete exosomes.15”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [15] Valadi, 2007, Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells, Nat. Cell Biol. source

  • May combine two claims.

    Interpretationown data

    Exosomes in serum of CUMS rats are mainly derived from microglia, which may enrich miR-146a-5p.

    “confirming that exosomes in serum of CUMS rats were mainly derived from microglia, which may enrich miR-146a-5p (Figure S8).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Study design and conductown data

    Markedly increased levels of circANKS1B in rats receiving AAV-circANKS1B and decreased levels in those receiving AAV-circANKS1B-shRNA verified high transfection efficiency in DG.

    “The markedly increased levels of circANKS1B in rats receiving AAV-circANKS1B and decreased levels in those receiving AAV-circANKS1B-shRNA verified that a high transfection efficiency was present in the DG (Figures S14D and S14E).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Exosomes can carry specific miRNAs to cell receptors and play a key role in the pathogenesis of central nervous system diseases.

    “Exosomes, which are considered as delivery vectors, can carry specific miRNAs to cell receptors, and play a key role in the pathogenesis of central nervous system diseases.36”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [36] Tsilioni, 2014, Exosomes in neurologic and psychiatric disorders, Clin. Ther. source

  • May combine two claims.

    Premisecitation

    miR-146a-5p is a microglial-enriched miRNA that affects excitatory synapses.

    “miR-146a-5p has been shown to be a microglial-enriched miRNA that affects excitatory synapses.17,18”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [17] McNeill, 2012, MicroRNAs shape the neuronal landscape, Neuron source

      Not checked in the free run.

    • cites [18] Prada, 2018, Glia-to-neuron transfer of miRNAs via extracellular vesicles: a new mechanism underlying inflammation-induced synaptic alterations, Acta Neuropathol. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Chronic unpredictable stress induces depression-like symptoms, which are accompanied with an inhibition of hippocampal DG neurogenesis in rats.

    “chronic unpredictable stress induces depression-like symptoms, which are accompanied with an inhibition of hippocampal DG neurogenesis in rats”
    Quote in the paperSubstantial claim
    • Rests on an assumption: CUMS-induced depression-like behavior in rats is a valid model of human depression.

  • Classified as probably not a substantial claim.

    Interpretationreasoning

    Microglia, which secrete exosomes containing miR-146a-5p, play an important role in the neurogenesis that is coupled with depression via the miR-146a-5p/KLF4 pathway.

    “Collectively, these data offer the novel hypothesis that microglia, which secrete exosomes containing miR-146a-5p, play an important role in the neurogenesis that is coupled with depression via the miR-146a-5p/KLF4 pathway (Figure 7).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: CUMS-induced depression-like behavior in rats is a valid model of human depression.

  • Classified as probably not a substantial claim.

    Premisecitation

    Glial-enriched miR-146a-5p can inhibit neuronal differentiation.

    “It has been reported that glial-enriched miR-146a-5p can inhibit neuronal differentiation.29”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [29] Jovicic, 2013, Comprehensive expression analyses of neural cell-type-specific miRNAs identify new determinants of the specification and maintenance of neuronal phenotypes, J. Neurosci. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    The dentate gyrus of the adult brain represents a key region for neurogenesis.

    “As the DG of the adult brain represents a key region for neurogenesis,37”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [37] Cameron, 2001, Adult neurogenesis produces a large pool of new granule cells in the dentate gyrus, J. Comp. Neurol. source

  • May combine two claims.

    Interpretationown data

    circANKS1B can function as a sequester for miR-146a-5p to regulate the translational expression of KLF4, thus participating in the neurogenesis associated with depression.

    “These results suggest that circANKS1B can function as a sequester for miR-146a-5p to regulate the translational expression of KLF4, thus participating in the neurogenesis associated with depression.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    miRNAs play important roles in neurogenesis and participate in the transition of neural stem cells from proliferation to differentiation stages.

    “miRNAs play important roles in neurogenesis and participate in the transition of NSCs from proliferation to differentiation stages.34”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [34] Marangon, 2019, MicroRNAs change the games in central nervous system pharmacology, Biochem. Pharmacol. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    miR-146a-5p suppresses neurogenesis in the DG region of the hippocampus, thereby enhancing depression-like behaviors.

    “miR-146a-5p suppresses neurogenesis in the DG region of the hippocampus, thereby enhancing depression-like behaviors.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Study design and conductown data

    miR-146a-5p expression at DG was significantly upregulated in normal rats or downregulated in CUMS rats after AAV-miR-146a-5p virus injection.

    “miR-146a-5p expression at this site was significantly upregulated in normal rats or downregulated in CUMS rats (Figures S12A and S12B).”
    Quote in the paperSubstantial claim

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

    Premisecitation

    The expression of KLF4 in neural stem cells can regulate the differentiation and migration of neurons in the developing cerebral cortex.

    “It has been reported that the expression of KLF4 in neural stem cells can regulate the differentiation and migration of neurons in the developing cerebral cortex.28”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [28] Cui, 2017, KLF4 knockdown attenuates TBI-induced neuronal damage through p53 and JAK-STAT3 signaling, CNS Neurosci. Ther. source

      Not checked in the free run.

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

    Study design and conductnone given

    All statistical tests were two-tailed and a p < 0.05 was required for results to be considered as statistically significant.

    “All statistical tests were two-tailed and a p < 0.05 was required for results to be considered as statistically significant.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    Injection sites were verified after behavioral tests and only data from rats with correct injection site placements were used for subsequent experiments.

    “Injection sites were verified after behavioral tests and only data from rats with correct injection site placements were used for subsequent experiments.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    miRNAs play a key role in post-transcriptional regulation and circRNAs function as miRNA sequesters to modulate the pathogenesis and progression of various diseases.

    “It is well known that miRNAs play a key role in post-transcriptional regulation and circRNAs function as miRNA sequesters to modulate the pathogenesis and progression of various diseases.32,52”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [32] Piwecka, 2017, Loss of a mammalian circular RNA locus causes miRNA deregulation and affects brain function, Science source
    • cites [52] Cheng, 2018, Circular RNA VMA21 protects against intervertebral disc degeneration through targeting miR-200c and X linked inhibitor-of-apoptosis protein, Ann. Rheum. Dis. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    circANKS1B mediates miR-146a-5p regulation of KLF4 mRNA expression and its functional involvement in the neurogenesis of depression.

    “we demonstrate that circANKS1B mediates miR-146a-5p regulation of KLF4 mRNA expression and its functional involvement in the neurogenesis of depression.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    KLF4 can specifically bind to the phosphorylated forms of STAT3 (p-STAT3) and inhibit its expression.

    “KLF4 can specifically bind to the phosphorylated forms of STAT3 (p-STAT3) and inhibit its expression.30”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [30] Gu, 2015, Mitochondrial E3 ligase March5 maintains stemness of mouse ES cells via suppression of ERK signalling, Nat. Commun. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    miR-146a-5p, enriched in microglia-derived exosomes, is shuttled to DG neurons, leading to inhibition of neurogenesis by affecting the target gene KLF4.

    “miR-146a-5p, which is enriched in microglia-derived exosomes, is shuttled to DG neurons, leading to an inhibition of neurogenesis by affecting the target gene, KLF4.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Exosomes can cross the blood-brain barrier.

  • May combine two claims.

    Interpretationreasoning

    miR-146a-5p binds with KLF4 to inhibit the expression of CDKL5 and increase STAT3 phosphorylation levels, thereby reducing neurogenesis and inducing depression-like behaviors.

    “In this study, miR-146a-5p appears to bind with KLF4 to inhibit the expression of CDKL5 and increase STAT3 phosphorylation levels, thereby reducing neurogenesis and inducing depression-like behaviors.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Abnormal expressions of circRNAs are present in the blood of patients with major depressive disorder.

    “abnormal expressions of circRNAs are present in the blood of MDD patients.51”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [51] Jiang, 2017, Relationships of circular RNA with diabetes and depression, Sci. Rep. source

      Not checked in the free run.

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

    Study design and conductnone given

    Normality between group samples was assessed using the D'Agostino and Pearson omnibus normality test and Brown-Forsythe tests.

    “Normality between group samples were assessed using the D'Agostino and Pearson omnibus normality test and Brown-Forsythe tests.”
    Quote in the paperNot a substantial claim
44 of 44

  • The paper relies on this without support.

    CUMS-induced depression-like behavior in rats is a valid model of human depression.

    “With use of the chronic unpredictable mild stress (CUMS)-induced rat model of depression”

  • The paper relies on this without support.

    Exosomes can cross the blood-brain barrier.

    “As exosomes can cross the blood-brain barrier, they offer a means for achieving a precise delivery of miRNA to its intended central nervous system target.”
2 of 2

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?

  • The link between miRNAs and the pathophysiology of depression may be implicated by the fact that miRNAs regulate neuronal regulation, neurotransmitter regulation, and microglia activation.

    “The link between miRNAs and the pathophysiology of depression248–252 may be implicated by the fact that miRNAs regulate neuronal regulation, neurotransmitter regulation, and microglia activation.253,254”

    From Epigenetic regulation in major depression and other stress-related disorders: molecular mechanisms, clinical relevance and therapeutic potential (2023) · cites this paper as [254]

    The Discussion (page unavailable) links microglia-derived miR-146a-5p to altered neurogenesis and neuronal function in depression. The Introduction mentions neurotransmitter changes as part of depression’s pathophysiology, but does not say miRNAs regulate neurotransmitters; the paper also does not establish that miRNAs regulate microglial activation. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • miR-146a-5p is transferred by microglia to the hippocampus DG, where it targets KLF4 and inhibits neurogenesis and spontaneous discharge of excitatory neurons.

    “miR-146a-5p is transferred by microglia to the hippocampus DG, where it targets Krüppel-like factor 4 (KLF4) and inhibits neurogenesis and the spontaneous discharge of excitatory neurons.254”

    From Epigenetic regulation in major depression and other stress-related disorders: molecular mechanisms, clinical relevance and therapeutic potential (2023) · cites this paper as [254]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Discussion identifies a different miR-146a isoform

    Minor

    The discussion calls the enriched, stem-cell-pathway-associated exosomal miRNA miR-146a-4p, whereas the corresponding results identify it as miR-146a-5p.

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Authors
Fan, C., Li, Y., Lan, T., Wang, W., Long, Y., & Yu, S. Y.
Affiliations
Shandong University (CN)
Corresponding author
Yu, Shu Yan
ORCID
Venue
Molecular Therapy
Publisher
Elsevier BV
Published
2022-03
Volume
30
Issue
3
Pages
1300-1314
ISSN
1525-0016 (ISSN-L)
DOI
10.1016/j.ymthe.2021.11.006
PMC
PMC8899528
OpenAlex
W3212530036
Semantic Scholar
6861e6855aaca44197cb811ed5090831b96e0fc6
Cited by
227
References
61
Citations by year
2026: 31 · 2025: 73 · 2024: 66 · 2023: 40 · 2022: 17
FWCI
11.20
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Therapy (publishedVersion, cc-by-nc-nd)
  • SSRN Electronic Journal (acceptedVersion)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • National Natural Science Foundation of China (NSFC81873796, 82071513)
  • Natural Science Foundation of Shandong Province
  • National Natural Science Foundation of China-Shandong Joint Fund for Marine Science Research Centers (ZR2020ZD25)
  • Fundamental Research Fund of Shandong University (2018JC008)
Topics
  • Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
Keywords
depression, microglia, exosomes, miR-146a, adult neurogenesis, dentate gyrus, KLF4, microRNA sequestration, post-transcriptional regulation
MeSH terms
Animals, Depression, Microglia, MicroRNAs, Rats, Exosomes, Neurogenesis

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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 2026-10-09.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-09.

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