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Cited by 31Neuroinflammation and Neurodegeneration MechanismsAlzheimer's disease research and treatmentsNeurological Disease Mechanisms and Treatments

A novel molecular class that recruits HDAC/MECP2 complexes to PU.1 motifs reduces neuroinflammation

Open access
Ralvenius, William T., Mungenast, Alison E., Woolf, Hannah, Huston, Margaret M., Gillingham, Tyler Z., Godin, Stephen K., Penney, Jay, Cam, Hugh P. +13 more·2023·Journal of Experimental Medicine
DOI 10.1084/jem.20222105PMC PMC10465325
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Abstract

Pervasive neuroinflammation occurs in many neurodegenerative diseases, including Alzheimer’s disease (AD). SPI1/PU.1 is a transcription factor located at a genome-wide significant AD-risk locus and its reduced expression is associated with delayed onset of AD. We analyzed single-cell transcriptomic datasets from microglia of human AD patients and found an enrichment of PU.1-binding motifs in the differentially expressed genes. In hippocampal tissues from transgenic mice with neurodegeneration, we found vastly increased genomic PU.1 binding. We then screened for PU.1 inhibitors using a PU.1 reporter cell line and discovered A11, a molecule with anti-inflammatory efficacy and nanomolar potency. A11 regulated genes putatively by recruiting a repressive complex containing MECP2, HDAC1, SIN3A, and DNMT3A to PU.1 motifs, thus representing a novel mechanism and class of molecules. In mouse models of AD, A11 ameliorated neuroinflammation, loss of neuronal integrity, AD pathology, and improved cognitive performance. This study uncovers a novel class of anti-inflammatory molecules with therapeutic potential for neurodegenerative disorders.

AI snapshot

Written from the paper’s full text.

TL;DR

The paper reports discovery of A11, a small molecule that inhibits PU.1 activity by recruiting MECP2/HDAC1-containing co-repressor complexes to PU.1 motifs, reducing neuroinflammation. In mouse models of AD-like pathology, A11 reduced microgliosis, astrogliosis, tau and amyloid pathology, and improved cognitive performance without observed hematopoietic side effects.

Key findings

  • From a screen of 58,100 small molecules, A11 was the most potent hit with an EC50 of 2.5 nM in the BV2 PU.1-Luciferase reporter assay, >16-fold more potent than any other hit; the final validated hit rate was 0.07% (43 hits).
  • In iMGLs, A11 (20 nM, 2 d) lowered IL1β mRNA (P ≤ 0.0478) and protein (P ≤ 0.0406), and reversed the direction of expression of ∼66% of 1,246 AD-associated immune genes in activated iMGLs. Mechanistically, A11 increased pulldown of MECP2 by ∼150% (P = 0.0050), HDAC1 by ∼266% (P = 0.0012), SIN3A by ∼267% (P = 0.0171), and DNMT3A by ∼124% (P = 0.0358) at PU.1 motifs, without changing PU.1 levels.
  • In CK-p25 mice, A11 reduced PU.1-positive nuclei by ∼47% (P = 0.0152) and IBA1 area by ∼54% (P = 0.0290) at 0.1 mg/kg; at 0.3 mg/kg it reduced C1q by ∼26% (P = 0.0152), GFAP by ∼46% (P = 0.0400), increased intact GFP-positive neurons by ∼29% (P = 0.0140), and increased tubulin β3 by ∼2.2-fold (P = 0.0038). After 6 wk, A11 improved Y-maze novel arm preference from −4.1 ± 9.4% to 18.7 ± 11.7% (P = 0.0007) and Morris water maze improvement from 3.5 ± 18.3% to 67.4 ± 30.1% (P = 0.0002).
  • In P301STau mice, A11 reduced phosphorylated tau by ∼56% (AT8, P = 0.0045) and ∼37% (Ser404, P = 0.0138), and improved Y-maze preference from 7.3 ± 8.0% to 25.7 ± 8.9% (P = 0.0008). In 5XFAD mice, A11 reduced hippocampal β-amyloid by ∼37% (P = 0.0356) and amyloid–microglia colocalization by ∼61% (P = 0.0424).

Methods

  • A BV2 PU.1-Luciferase reporter line with five tandem λB PU.1 motifs was used to screen 58,100 small molecules twice at 3.3 µM, followed by four validation filters; hits were tested in human iPSC- and ES-derived iMGLs using uptake assays, RT-qPCR, ELISA, bulk RNA-seq, ChIP-qPCR, EMSA, streptavidin pulldown, and Western blotting.
  • In vivo studies used CK-p25, P301STau, and 5XFAD mice; A11 was injected intraperitoneally at 0.1–1 mg/kg or 0.3 mg/kg daily for 2 or 6 wk, followed by immunohistology, FACS of microglia, bulk RNA-seq, behavioral testing (Y maze, Morris water maze), and pharmacokinetic analysis in C57BL/6J mice.
  • Transcriptomic analyses used DESeq2/apeglm for differential expression, HOMER for motif enrichment, and gene ontology tools; statistical comparisons used t tests or ANOVA with post hoc tests as reported.

Limitations

  • The paper states that the mouse models are imperfect: no single strain exhibits all AD hallmarks, transgenic mice lack human genetic diversity and cannot model decades of human aging, and younger mice will be needed to test preventative use. It also states that long-term studies in higher organisms (e.g., rats, dogs, primates) and clinical studies in AD patients are required before hematological or peripheral side effects can be fully excluded.
  • Evident from the text: several in vivo effects are trends or borderline (e.g., P301STau PU.1-positive nuclei P = 0.0874; 6-wk CK-p25 IBA1 P = 0.0503 and C1q P = 0.0534; HDAC1 ChIP at Il1b P ≤ 0.08 and Ccl2 P < 0.2), and some experiments used only one sex (e.g., 2-wk CK-p25 immunohistology in males, transcriptomics in females; 6-wk CK-p25 in females; P301STau and 5XFAD in males), limiting generalizability.

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Integrity signals

Registry

Crossref · 10.1084/jem.20222105

Cites

79 of 79 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

47 of 57 cited sources checked · 19 supported · 1 flagged · accuracy not yet measured

  • Premisecitation

    Alzheimer's disease risk alleles are enriched in PU.1-binding motifs.

    “Notably, AD risk alleles were recently shown to be enriched in PU.1-binding motifs (Novikova et al., 2021).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Novikova et al., 2021 source
  • Premisecitation

    Total inhibition of PU.1 activity could adversely interfere with myeloid differentiation.

    “While total inhibition of PU.1 activity could adversely interfere with myeloid differentiation (Iwasaki et al., 2005)”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites Iwasaki et al., 2005 source
  • Interpretationown data

    A11 has favorable structural properties for CNS drug development.

    “Additionally, A11 scores 4.8 out of 6 on the central nervous system multiparameter optimization (CNS MPO; Wager et al., 2016) desirability index, indicating favorable structural properties for CNS drug development (Fig. S4 d).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Wager et al., 2016 source
  • Premisecitation

    The proteins MECP2, HDAC1, SIN3A, and DNMT3A that were increased after A11 treatment belong to transcriptional co-repressor complexes known to interact with PU.1.

    “The proteins MECP2, HDAC1, SIN3A, and DNMT3A that were increased after A11 treatment belong to transcriptional co-repressor complexes known to interact with PU.1 (Gregoricchio et al., 2022; Hallier et al., 1996).”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites Gregoricchio et al., 2022 source

      The Results section (page unavailable) reports that SPI1/PU.1, HDAC1 and SIN3A appear to belong to the same complex, supporting the claim for HDAC1 and SIN3A. Our copy does not establish that MECP2 or DNMT3A belong to PU.1-interacting co-repressor complexes; it mentions DNMT3A only in the Introduction as a gene commonly mutated in AML. The increase after A11 treatment is the citing authors’ observation, not a finding attributed to this source. (LLM, conf=0.90)

      Experimental — accuracy not yet measured
    • cites Hallier et al., 1996 source

      The Results and Discussion report that PU.1 associates with the RNA-binding protein p54nrb, not with a transcriptional co-repressor complex. Searches for MECP2, HDAC1, SIN3A, DNMT3A and co-repressor terms found no passage in our copy linking those proteins or complexes to PU.1. The reported increase after A11 treatment is not established by this source either. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
  • Interpretationreasoning

    A11 treatment could affect inflammation-induced changes in microglial adhesion and motility.

    “suggesting that A11 treatment could affect inflammation-induced changes in microglial adhesion and motility (Socodato et al., 2020).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Socodato et al., 2020 source

      The Introduction (p. 2) links proinflammatory microglial activation to cytoskeletal changes, and “Rhoa Is Required for Microglial Survival and Activation” (p. 4) reports that inhibiting Rhoa slows microglial protrusion extension. The paper does not examine A11 treatment or establish an effect on microglial adhesion, so it supports only the broader connection between inflammation, cytoskeletal dynamics, and movement-related behavior. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
  • Classified as probably not a substantial claim. 32
  • May combine two claims. 48
  • The quote only partly states this claim. 5

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

    Premisecitation

    Association between PU.1 and this repressive complex has previously been suggested to represent an endogenous defense mechanism in erythroid cells that restricts leukemia-associated gene expression and survival.

    “Association between PU.1 and this repressive complex has previously been suggested to represent an endogenous defense mechanism in erythroid cells that restricts leukemia-associated gene expression and survival (Gregoricchio et al., 2022; Suzuki et al., 2003).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites Gregoricchio et al., 2022 source

      The Introduction says elevated PU.1 expression favors erythroid-progenitor survival and initiates erythroleukaemia. The Results describe HDAC1 and PRC2 as mediating PU.1 repression that enables leukaemic-cell survival; inhibiting the complexes instead reduces survival. This is contrary to the claimed defensive role. (LLM, conf=0.91)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites Suzuki et al., 2003 source

  • May combine two claims.

    Premisecitation

    Increased PU.1 expression levels are associated with Alzheimer's disease and Huntington's disease.

    “Increased PU.1 expression levels are associated with AD (Gjoneska et al., 2015; Huang et al., 2017; Rustenhoven et al., 2018) and Huntington’s disease (Crotti et al., 2014)”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites Gjoneska et al., 2015 source

      The paper reports increased PU.1 (SPI1) expression in a CK-p25 mouse model of AD-like neurodegeneration (passage [1], section labeled Abstract). That supports the AD association, but does not establish increased PU.1 expression in people with AD. The Huntington’s disease portion of the citing sentence is attributed to Crotti et al., not to this paper. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites Huang et al., 2017 source
    • cites Rustenhoven et al., 2018 source

  • May combine two claims.

    Premisecitation

    Polymorphisms in the PU.1 gene SPI1 that reduce PU.1 expression levels lower the risk of Alzheimer's disease and delay Alzheimer's disease onset.

    “while polymorphisms in the PU.1 gene SPI1 that reduce PU.1 expression levels lower the risk of AD (Cao et al., 2022) and delay AD onset (Huang et al., 2017).”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites Cao et al., 2022 source
    • cites Huang et al., 2017 source

  • May combine two claims.

    Premisecitation

    A majority of Alzheimer's disease risk genes identified through genome-wide association studies are highly expressed in microglia, suggesting a key role in microglial dysfunction.

    “A majority of AD risk genes identified through genome-wide association studies are highly expressed in microglia, the brain’s innate immune cells, suggesting a key role in microglial dysfunction (Kummer et al., 2021; Penney et al., 2020; Teipel et al., 2022).”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites Kummer et al., 2021 source

      The Results section ('PD-1 regulates microglial Aβ uptake and inflammation'; page unavailable) links altered microglial gene expression to inflammatory pathways, supporting a role for microglial dysfunction in AD pathology. It does not say that a majority of GWAS-identified AD risk genes are highly expressed in microglia; searches for 'genome-wide association studies' and 'GWAS risk genes' found no such passage. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites Penney et al., 2020 source

      The Microglia section (page unavailable) says GWAS identified a surprising number of microglial genes as late-onset AD risk factors and links those findings to microglial dysfunction. It does not establish that a majority of GWAS-identified AD risk genes are highly expressed in microglia; the Alzheimer’s disease section instead says many are expressed primarily in non-neuronal brain cells. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites Teipel et al., 2022 source

      The Introduction discusses microglia-derived inflammatory factors in Alzheimer’s disease, and the Discussion addresses markers of microglial activation, but neither addresses expression of AD risk genes in microglia. Searches for genome-wide association studies and microglial expression of genetic risk genes found no passage supporting the claimed majority. (LLM, conf=0.75)

      Experimental — accuracy not yet measured

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

    Premisecitation

    Microglial activation can help clear pathogenic molecules from the brain but can also lead to chronic or excessive inflammation that exacerbates neurodegeneration.

    “While microglial activation can help clear pathogenic molecules from the brain, it can also lead to chronic or excessive inflammation that exacerbates neurodegeneration (Mathys et al., 2017).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites Mathys et al., 2017 source

      The Introduction says microglia clear apoptotic cells, but it does not say they clear pathogenic molecules. The Discussion reports increased pro-inflammatory cytokines in responding microglia and suggests they may contribute to a cascade leading to neuronal death; it does not establish that chronic or excessive inflammation exacerbates neurodegeneration. (LLM, conf=0.85)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Interpretationcitation

    The enrichment of ETS, IRF, bHLH, and bZIP motifs suggests a central role for PU.1, a member of the ETS family and known binding partner of other ETS proteins, and IRF, bHLH, and bZIP transcription factors.

    “This suggested a central role for PU.1, a member of the ETS family and known binding partner of other ETS proteins, and IRF, bHLH, and bZIP transcription factors (Gupta et al., 2009).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites Gupta et al., 2009 source

      The ‘Transcription factor PU.1: expression distribution and function’ section identifies PU.1 as an ETS-family member, and ‘Functional domains of PU.1 protein’ reports interactions with IRFs and c-Jun; the ‘NF-IL6β (C/EBP-δ)’ section reports interaction with a leucine-zipper factor. The passages do not establish PU.1 as a binding partner of other ETS or bHLH factors. The inference from motif enrichment is the citing authors’ own, rather than a finding attributed to this review. Page numbers were not supplied in our copy. (LLM, conf=0.84)

      Experimental — accuracy not yet measured

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

    Premisecitation

    In brain tissue from human Alzheimer's disease patients, microglia display the morphology and gene expression profile characteristic of an activated cell state and cluster around β-amyloid plaques.

    “In brain tissue from human AD patients, microglia display the morphology (Heppner et al., 2015) and gene expression profile (Gjoneska et al., 2015; Mathys et al., 2019) characteristic of an activated cell state and cluster around β-amyloid plaques.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites Heppner et al., 2015 source
    • cites Gjoneska et al., 2015 source

      The paper reports higher immune-gene expression in hippocampal tissue from human AD patients and an activated microglial gene-expression signature in its mouse model (main text, page unavailable). It does not establish an activated gene-expression profile specifically in microglia isolated from human patients. The sentence cites Heppner et al. for morphology, not this paper; this paper also does not establish microglial clustering around plaques. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites Mathys et al., 2019 source

  • May combine two claims.

    Premisecitation

    PU.1 is a lineage-specifying transcription factor most highly expressed in peripheral myeloid cells and constitutively in microglia, where it can drive inflammatory gene expression together with IRFs and NF-κB.

    “The transcription factor PU.1 is a lineage-specifying transcription factor that is most highly expressed in peripheral myeloid cells and constitutively in microglia in the brain, where it can drive inflammatory gene expression together with other transcription factors such as the IFN regulatory factors (IRFs) and NF-κB (Gupta et al., 2009).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites Gupta et al., 2009 source

      The ‘expression distribution and function’ and ‘PU.1 gene regulation’ sections describe PU.1 as a regulator of myeloid lineage fate, with high expression in monocytic and granulocytic lineages. The ‘Functional domains’ and ‘PU.1-interacting proteins’ sections describe interactions with IRFs and cooperation with other factors in inflammatory-gene transcription. Our copy does not establish that expression is highest specifically in peripheral myeloid cells, that PU.1 is constitutively expressed in brain microglia, or that it drives inflammatory genes together with NF-κB (no page numbers supplied). (LLM, conf=0.88)

      Experimental — accuracy not yet measured

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

    Premisecitation

    Our results are in line with studies showing that reduced microglial colocalization with β-amyloid is associated with a reduced β-amyloid burden.

    “Our results are in line with studies showing that reduced microglial colocalization with β-amyloid is associated with a reduced β-amyloid burden (D’Errico et al., 2022; Spangenberg et al., 2019).”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites D’Errico et al., 2022 source

      The Main section (page not provided) reports less microglial clustering around Aβ plaques and fewer amyloid deposits in wild-type grafts when microglial migration into those grafts was reduced. But the mice had similar numbers of Aβ plaques in the surrounding host tissue, so the finding supports reduced graft deposition, not a general reduction in Aβ burden. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites Spangenberg et al., 2019 source

      The Results section (“Microglial elimination impairs plaque formation”; page unavailable) reports fewer parenchymal Aβ plaques in microglia-depleted 5xFAD mice, supporting the association if ‘burden’ means plaque burden. The same section says soluble and insoluble Aβ levels did not significantly change: depletion shifted where Aβ accumulated rather than reducing its net amount. The source therefore does not support a reduction in overall Aβ burden. (LLM, conf=0.87)

      Experimental — accuracy not yet measured

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

    Premisecitation

    Microglial depletion has been shown to reduce plaque burden but also alter the morphology of plaques.

    “With regards to amyloid-β, microglial depletion has been shown to reduce plaque burden but also alter the morphology of plaques (Casali et al., 2020).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Casali et al., 2020 source

  • May combine two claims.

    Premisecitation

    In the same study, an increase in plaque-associated tau in dystrophic neurites was found, indicating that microglia play a complex role in both the spread of tau and clearance of neuronal debris associated with pathological tau.

    “However, in the same study, an increase in plaque-associated tau in dystrophic neurites was found, indicating that microglia play a complex role in both the spread of tau and clearance of neuronal debris associated with pathological tau.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Clayton et al., 2021 source

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

    Premisecitation

    AD risk variants are enriched in genes that are highly expressed in microglia.

    “Population genetics has revealed that AD risk variants are enriched in genes that are highly expressed in microglia (Penney et al., 2020).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Penney et al., 2020 source

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

    Premisecitation

    AD is characterized by neuroinflammation driven by activated microglia, a phenotype shared by many other forms of neurodegeneration.

    “In addition to β-amyloid and hyperphosphorylated tau accumulation in the brain, a major hallmark of AD is that of neuroinflammation driven by activated microglia, a phenotype shared by many other forms of neurodegeneration (Gjoneska et al., 2015).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Gjoneska et al., 2015 source

      The paper’s main text (indexed under Abstract; page unavailable) reports increased immune-gene expression in human AD samples and a significant contribution from microglial activation in an AD-like mouse model. It does not establish that activated microglia drive neuroinflammation in human AD or that this phenotype is shared by many other forms of neurodegeneration. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Interpretationcitation

    The beneficial effects of A11 might stem from a reduction in PU.1 activity below a threshold that otherwise causes microglia to acquire a detrimental and hyperinflammatory phenotype or that helps undo unresolved microglial activation.

    “The beneficial effects of A11 might stem from a reduction in PU.1 activity below a threshold that otherwise causes the microglia to acquire a detrimental and hyperinflammatory phenotype (Crotti et al., 2014; Gjoneska et al., 2015) or that helps the undoing of unresolved microglial activation (Leng and Edison, 2021).”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites Crotti et al., 2014 source

      The Results report that knocking down PU.1 greatly reduced IL-6 and TNF-α expression in mutant-huntingtin microglia, and the Discussion links increased PU.1 to pro-inflammatory and neurotoxic activity. Crotti et al. do not establish a PU.1 activity threshold or show that crossing one causes microglia to acquire that phenotype. The proposed benefit of A11 is the citing authors’ inference, while the unresolved-activation clause is attributed to a different source. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites Gjoneska et al., 2015 source

      The paper’s Abstract reports increased PU.1 expression in the neurodegeneration model, links PU.1 to microglial activation, and proposes it as a therapeutic target. It does not establish a PU.1 activity threshold or show that crossing one causes microglia to acquire a detrimental, hyperinflammatory phenotype. The A11 effect and reversal of unresolved activation are not attributed to Gjoneska et al. in the citing sentence. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites Leng and Edison, 2021 source

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

    Premisecitation

    Microglia have been suggested to deposit plaques in non-burdened brain tissue.

    “Microglia have as well been suggested to deposit plaques in non-burdened brain tissue (D’Errico et al., 2022).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites D’Errico et al., 2022 source

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

    Premisecitation

    Amyloid-β-associated microglia secrete extracellular vesicles that contain phosphorylated tau, and their depletion abrogated tau propagation.

    “Amyloid-β–associated microglia were shown to secrete extracellular vesicles that contain phosphorylated tau and their depletion abrogated tau propagation (Clayton et al., 2021).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Clayton et al., 2021 source

      The Discussion (‘Microglia depletion and tauopathy’; page unavailable) reports that plaque-associated microglia secrete extracellular vesicles containing phosphorylated tau. The Results (‘Enhanced tau propagation and its reduction by CSF1R inhibition in AppNL-G-F mice’; page unavailable) say microglial depletion effectively suppressed tau propagation in the mouse model, but do not establish that it abrogated propagation entirely. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    Previous studies have shown that microglial depletion can have beneficial effects in transgenic mice that model AD hallmarks, including reduction in brain accumulation of both amyloid-β and tau.

    “In support of this interpretation, previous studies have shown that microglial depletion can have beneficial effects in transgenic mice that model AD hallmarks, including reduction in brain accumulation of both amyloid-β and tau (Sosna et al., 2018; Spangenberg et al., 2019).”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites Sosna et al., 2018 source

      The Results section reports that early, long-term PLX3397 treatment depleted microglia and reduced intraneuronal amyloid and amyloid plaques in 5XFAD mice. The cited paper does not report a reduction in tau accumulation, so it supports the amyloid-β portion of the combined claim but not the tau portion. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites Spangenberg et al., 2019 source

      The Results section ('Microglial elimination impairs plaque formation'; page unavailable) reports fewer cortical amyloid-β plaques in microglia-depleted 5xFAD mice, supporting the amyloid-plaque part of the claim. It also says total amyloid-β amounts were unchanged and accumulation shifted toward blood vessels, so it does not support reduced overall brain accumulation. The passages found do not report reduced tau accumulation. (LLM, conf=0.89)

      Experimental — accuracy not yet measured

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

    Premisecitation

    Studies using transgenic P301STau mice report that microglial depletion reduced neurodegeneration and the accumulation of phosphorylated tau.

    “In line with our findings, studies using transgenic P301STau mice report that microglial depletion reduced neurodegeneration and the accumulation of phosphorylated tau (Johnson et al., 2023; Mancuso et al., 2019; Shi et al., 2019).”
    Quote in the paperNot a substantial claim3 of 3 sources checked
    • cites Johnson et al., 2023 source

      The Results section on pathogenic tau reports that CSF1R-inhibitor treatment reduced phosphorylated tau in P301S Tg2541 mice. But the Results section on CNS injury reports reduced plasma NfL, a marker of neuronal injury, in treated females and increased NfL in treated males under the continuous-treatment regimen. Thus, the unqualified claim that microglial depletion reduced neurodegeneration omits an important sex-dependent limitation. No page numbers are available in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites Mancuso et al., 2019 source
    • cites Shi et al., 2019 source

  • May combine two claims.

    Findingown data

    A11 had an EC50 of 2.5 nM in the BV2 reporter assay and was more than 16-fold more potent than any other hit.

    “With an EC50 of 2.5 nM, A11 (Fig. 1, g and h) was >16-fold more potent in the BV2 reporter assay than any other hit.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Of the 2,060 upregulated PU.1 target genes in CK-p25 mice, A11 reduced the expression of 8.5% and further upregulated only 1.5%.

    “Of the 2,060 upregulated PU.1 target genes, A11 reduced the expression of 8.5% of the genes. In comparison, only 1.5% were further upregulated, suggesting that A11 predominantly ameliorates the activation of PU.1 target genes in CK-p25 mice (Fig. 3 i).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In female CK-p25 mice injected with A11 for 6 wk, there was a reduction in PU.1 positive nuclei by ∼25% (P = 0.0449), IBA1 immunoreactivity by ∼28% (P = 0.0503), C1q-positive area by ∼15% (P = 0.0534), and GFAP-positive area by ∼25% (P = 0.0213).

    “When simultaneously injecting female CK-p25 mice with A11 for 6wk, we again saw a reduction in the number of PU.1 positive nuclei by ∼25% (P = 0.0449), in IBA1 immunoreactivity by ∼28% (P = 0.0503), in C1q-positive area by ∼15% (P = 0.0534), and in GFAP-positive area by ∼25% (P = 0.0213).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    ChIP-qPCR on whole brain homogenate from A11-injected CK-p25 mice found strong trends or significant increases in HDAC1 signal for Il1b (P ≤ 0.08), Tnf (P ≤ 0.04), and Ccl2 (P < 0.2).

    “We found strong trends or significant increases in HDAC1 signal for the PU.1 target genes Il1b (P ≤ 0.08), Tnf (P ≤ 0.04), and Ccl2 (P < 0.2; Fig. 3 k).”
    Quote in the paperSubstantial claim

  • May combine two claims. The quote only partly states this claim.

    Findingown data

    In female CK-p25 mice injected with A11 for 6 wk, neuronal structures were preserved, seen as ∼1.9-fold increase of pyramidal layer tubulin β3 staining (P = 0.0440) and ∼1.4-fold increase in intact GFP-positive cells in the granule layer (P = 0.0288).

    “Neuronal structures were also preserved, as seen by an ∼1.9-fold increase of pyramidal layer tubulin β3 staining (P = 0.0440) and a ∼1.4-fold increase in intact GFP-positive cells in the granule layer (P = 0.0288; Fig. 4 e; for effect in CK mice, see Table S12).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In 1-yr-old male 5XFAD mice, A11 treatment caused a reduction in hippocampal β-amyloid accumulation by ∼37% (P = 0.0356) and colocalization of amyloid with microglia by ∼61% (P = 0.0424).

    “A11 treatment caused a reduction in both hippocampal β-amyloid accumulation by ∼37% (P = 0.0356) and colocalization of amyloid with microglia by ∼61% (P = 0.0424; Fig. 4 h).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    A11 treatment improved performance of CK-p25 mice in the Y maze, increasing novel arm preference above chance from −4.1 ± 9.4% in vehicle-injected mice to 18.7 ± 11.7% in A11-injected mice (P = 0.0007).

    “A11 treatment improved performance of CK-p25 mice in the Y maze (Cruz et al., 2003; Fischer et al., 2005), increasing the novel arm preference above chance from −4.1 ± 9.4% in vehicle-injected mice to 18.7 ± 11.7% to in A11-injected mice (P = 0.0007; left panel of Fig. 4 i).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Cruz et al., 2003 source
    • cites Fischer et al., 2005 source

  • May combine two claims.

    Findingown data

    PU.1 binding was strongly increased in the hippocampus of male 6-wk induced CK-p25 mice, with more than 40% of all PU.1 ChIP peaks showing increased occupancy and only 0.1% showing decreased occupancy.

    “The ChIP-Seq revealed that PU.1 binding was strongly increased in the hippocampus of male 6-wk induced CK-p25 mice, with more than 40% of all PU.1 ChIP peaks showing increased occupancy and only 0.1% showing decreased occupancy (Fig. 1 b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    A11 prevented the appearance of a mobility shift for the PU.1 motif probes in a concentration-dependent manner, without affecting the EBNA probe which lacks the PU.1 motif.

    “Adding A11 to the binding reactions prevented the appearance of a mobility shift for the PU.1 motif probes in a concentration-dependent manner, without affecting the EBNA probe which lacks the PU.1 motif (lanes 5–14; Fig. 3, a and b)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    After a single systemic injection of A11 (0.3 mg/kg, i.p.) in wild-type mice, A11 was barely detected in plasma, had similar peak concentrations in peripheral fat and brain (cmax = 131 and 103 nM), and was eliminated three times faster from peripheral fat than brain (t1/2 = 1.1 and 3.7 h).

    “Strikingly, A11 was barely detected in plasma, and while present at similar peak concentrations in peripheral fat and brain (cmax = 131 and 103 nM, respectively), A11 was eliminated three times faster from peripheral fat compared with the brain (t1/2 = 1.1 and 3.7 h, respectively; Fig. 4 a).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In 1-yr-old male P301STau mice, daily A11 treatment (0.3 mg/kg, i.p.) for 6 wk reduced PU.1-positive nuclei by ∼61% (P = 0.0874), IBA1-positive area by ∼58% (P = 0.0132), C1q-positive area by ∼70% (P = 0.0038), and GFAP-positive area by ∼73% (P = 0.0104).

    “Daily A11 treatment (0.3 mg/kg, intraperitoneally) for 6 wk of 1-yr-old male P301STau mice had similar anti-inflammatory effects—reduction in the number of PU.1-positive nuclei by ∼61% (P = 0.0874), in IBA1-positive area by ∼58% (P = 0.0132), in C1q-positive area by ∼70% (P = 0.0038), in GFAP-positive area by ∼73% (P = 0.0104)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    A11 reduced microglial activation in the dentate gyrus of CK-p25 mice at 0.1 mg/kg, seen as ∼47% reduction in PU.1-positive nuclei (P = 0.0152) and ∼54% reduction in IBA1-positive surface area (P = 0.0290).

    “A11 reduced microglial activation in the dentate gyrus of CK-p25 mice at 0.1 mg/kg, seen as a ∼47% reduction in the number of PU.1-positive nuclei (P = 0.0152) and a ∼54% reduction in IBA1-positive surface area (P = 0.0290).”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    The primary screen of 58,100 small molecules yielded 264 hits.

    “The primary screen yielded 264 hits that were subject to four additional filters for inhibition of PU.1 activity.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In P301STau mice, A11 treatment reduced phosphorylated tau by ∼56% (AT8, P = 0.0045) and ∼37% (Ser404, P = 0.0138).

    “The P301STau mice also exhibited a reduction in phosphorylated tau of ∼56% (P = 0.0045) as measured with the AT8 antibody and ∼37% (P = 0.0138) as measured with the Ser404 antibody after A11 treatment (Fig. 4 g; for effect in littermates, see Table S13).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingcitation

    Recombinant PU.1 alone did not bind the λB probe, which requires posttranslational modifications of PU.1.

    “Recombinant PU.1 alone did not bind the λB probe (Fig. S3 c and SourceData FS3 e), which requires posttranslational modifications of PU.1 (Eisenbeis et al., 1993).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Eisenbeis et al., 1993 source

  • May combine two claims.

    Findingown data

    Compared with vehicle, there was ∼29% more intact GFP-positive p25-expressing neurons remaining in the granule layer after 0.3 mg/kg A11 (P = 0.0140) and ∼2.2-fold more tubulin β3 immunoreactivity in the pyramidal layer (P = 0.0038).

    “Compared with vehicle-treated mice, there was ∼29% more intact GFP-positive p25-expressing neurons remaining in the granule layer after the 0.3 mg/kg A11-treatment (P = 0.0140) as well as ∼2.2-fold more tubulin β3 immunoreactivity in the pyramidal layer (P = 0.0038; Fig. 4 d).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Hematopoietic stem cell expression of CD43 (P > 0.1141), CD11b (P > 0.1868), and proliferation (P > 0.4836) were unaffected by up to 5 d of A11 exposure.

    “Flow cytometry experiments showed that hematopoietic stem cell expression of CD43 (P > 0.1141), CD11b (P > 0.1868), and proliferation (P > 0.4836) were unaffected at various points throughout maturation by up to 5 d of A11 exposure (Fig. S5, a–c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    A11 did not affect peripheral hematopoiesis or cause other side effects.

    “After several rounds of validation, we discovered the novel small molecule A11, a promising therapeutic that reduced inflammation, mitigated hallmarks of AD, and improved cognitive function across multiple mouse models, without affecting peripheral hematopoiesis or causing other side effects.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Mice injected with A11 daily for 6 wk did not show any difference in circulating peripheral CD45+, CD4+, and CD8+ cells, nor any liver or spleen anomalies.

    “Mice injected with A11 daily for 6 wk also did not show any difference in circulating peripheral CD45+, CD4+, and CD8+ cells, nor any liver or spleen anomalies.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Targeting PU.1 therapeutically carries potential for alterations in hematopoiesis.

  • May combine two claims.

    Findingown data

    Chronic A11 treatment reduced accumulation of hyperphosphorylated tau in P301STau mice and β-amyloid in 5XFAD mice.

    “Chronic A11 treatment also reduced accumulation of hyperphosphorylated tau in P301STau mice and accumulation of β-amyloid in 5XFAD mice.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    A11-treated P301STau mice showed a significant improvement in spatial memory retention by 58.9 ± 31.0% compared with the first trial day, while vehicle-treated mice only improved by 20.6 ± 25.0% (P = 0.0168).

    “A11-treated P301STau mice also showed a significant improvement in spatial memory retention by 58.9 ± 31.0% compared with the first trial day, while vehicle-treated mice only improved by 20.6 ± 25.0% (P = 0.0168; middle and right panel of Fig. 4 j).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    On the final trial day, A11-treated CK-p25 mice showed a 67.4 ± 30.1% reduction in time required to find the submerged platform compared with a 3.5 ± 18.3% reduction in vehicle-treated CK-p25 mice (P = 0.0002), indicating improved spatial memory retention.

    “Performance on the final trial day, quantified as percent improvement versus the first trial day, showed a 67.4 ± 30.1% reduction in A11-treated CK-p25 mice compared with a 3.5 ± 18.3% reduction in vehicle-treated CK-p25 mice in time required to find the submerged platform, indicating that A11 treatment significantly improved spatial memory retention (P = 0.0002; right panel of Fig. 4 i).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In the Morris Water Maze, A11-treated CK-p25 mice performed similarly to CK littermates, requiring fewer seconds to locate the submerged platform on each consecutive trial day, while vehicle-treated CK-p25 mice showed impairment.

    “During the initial trial days, A11-treated CK-p25 mice appeared to perform similarly to the CK littermates, requiring fewer seconds to locate the submerged platform on each consecutive trial day, while vehicle-treated CK-p25 mice showed impairment (middle panel of Fig. 4 i).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In wild-type male mice injected with A11 for 6 wk, there was no change in the ratio of circulating CD45 (P = 0.6203), CD4 (P = 0.7111), or CD8-positive cells (P = 0.3722) and no abnormalities in liver or spleen histology.

    “We also investigated blood cells from wild-type male mice injected with A11 for 6 wk (as in Fig. 4 e), finding no change in the ratio of circulating CD45 (P = 0.6203), CD4 (P = 0.7111), or CD8-positive cells (P = 0.3722) and no abnormalities in liver or spleen histology (Fig. S5, d and e).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Up to 1 μM A11 had no effect on the proliferation or viability of human iPSC-derived hematopoietic stem cells or human erythroid or bone marrow-derived progenitors (EC50 > 1 μM).

    “Additionally, up to 1 µM A11 (equivalent to 500 times the cellular potency in the BV2 PU.1 luciferase assay) had no effect on the proliferation or viability of human iPSC–derived hematopoietic stem cells or in human erythroid or bone marrow-derived progenitors (EC50 > 1 µM; Fig. S5, f and g).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In P301STau mice, A11 treatment increased novel arm preference in the Y maze from 7.3 ± 8.0% (vehicle) to 25.7 ± 8.9% (P = 0.0008), without effect in wild-type littermates (vehicle: 30.2 ± 13.6% and A11: 30.7 ± 8.3%, P = 0.9).

    “Similarly, vehicle-treated P301STau mice exhibited a 7.3 ± 8.0% novel arm preference while A11 treatment resulted in a 25.7 ± 8.9% preference (P = 0.0008), without effect in wild-type littermates (vehicle: 30.2 ± 13.6% and A11: 30.7 ± 8.3% above chance, P = 0.9; left panel of Fig. 4 j).”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Neuroinflammation is a common feature of Alzheimer's disease and may be a potential treatment target.

    “Widespread neuroinflammation is a common feature of AD and may present a potential treatment target (Koutsodendris et al., 2021).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Koutsodendris et al., 2021 source

  • May combine two claims.

    Interpretationown data

    These data suggest that A11 should have promising distribution in the brain while being cleared rapidly from plasma.

    “These data suggest that A11 should have promising distribution in the brain while being cleared rapidly from plasma.”
    Quote in the paperSubstantial claim

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

    Study design and conductown data

    The non-specific luciferase quencher 119113 reduced luminescence by ∼90% in both BV2 PU.1-Luciferase and HEK CMV-Luciferase cells.

    “The non-specific luciferase quencher 119113 reduced luminescence by ∼90% in both BV2 PU.1-Luciferase and HEK CMV-Luciferase cells (P ≤ 0.0001; Table S5).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    The mobility shift only occurred with nuclear lysates from iMGLs and not from other cell types, suggesting the involvement of PU.1 binding to the λB probe.

    “The shift only occurred with nuclear lysates from iMGLs and not from other cell types, suggesting the involvement of PU.1 binding to the λB probe (Fig. S3 a and SourceData FS3 b)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    A11 readily crosses the blood-brain barrier and is rapidly eliminated from blood and peripheral fat, concentrating its exposure to brain-resident cells.

    “Thus, A11 readily crosses the blood–brain barrier and is rapidly eliminated from blood and peripheral fat, concentrating its exposure to brain-resident cells.”
    Quote in the paperSubstantial claim

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

    Interpretationnone given

    For immunohistochemical experiments with 2-wk induced CK-p25 mice and CK littermates, only males were used.

    “For immunohistochemical experiments with 2-wk induced CK-p25 mice and CK littermates, only males were used.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Transgenic mice lack the genetic diversity of human AD patients and cannot model the many decades that span a human lifetime.

    “We also note that transgenic mice lack the genetic diversity of human AD patients and simply cannot model the many decades that span a human lifetime.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). The quote only partly states this claim.

    Study design and conductown data

    Results in Figure 4 panels were repeated in two separate batches of experiments.

    “Results in this panel were repeated in two separate batches of experiments.”
    Quote in the paperNot a substantial claim

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

    Interpretationnone given

    For transcriptomic experiments with 2-wk induced CK-p25 mice and CK littermates, only females were used.

    “For transcriptomic experiments with 2-wk induced CK-p25 mice and CK littermates, only females were used.”
    Quote in the paperNot a substantial claim

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

    Study design and conductreasoning

    Effective AUC calculation measures luminescence inhibition without possible confounding loss of luminescence caused by cell death.

    “Calculations of the effective AUC were performed by taking the area under the luminescence inhibition curve minus the area under the Hoechst signal curve, which gives a measure of luminescence inhibition without the possible confounding loss of luminescence caused by cell death.”
    Quote in the paperNot a substantial claim

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

    Premisereasoning

    p25 expression is suppressed by a constant 1 g/kg doxycycline diet and induced when mice are put on a regular diet.

    “Therefore, p25 expression is suppressed by a constant 1 g/kg doxycycline diet (Bio-Rad, custom-made) and induced when mice are put on a regular diet (Teklad RMH 3000; Envigo).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Cruz et al., 2003 source

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

    Interpretationnone given

    All CK-p25:CX3CR1CreERT2/+ mice that were analyzed in Fig. 1 c were males.

    “All CK-p25:CX3CR1CreERT2/+ mice that were analyzed in Fig. 1 c were males.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    EMSA was repeated in four separate batches of experiments.

    “EMSA was repeated in four separate batches of experiments.”
    Quote in the paperNot a substantial claim

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

    Interpretationreasoning

    These discrepancies might be explained by the tau pathology model used.

    “These discrepancies might be explained by the tau pathology model used.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    In CK-p25 mice, p25 expression is driven by the CaMKII-tTA (CK) promoter crossed to tetO-p25 mice in the absence of doxycycline, with CK mice as littermate controls.

    “In CK-p25 mice, p25 expression is driven by the CaMKII-tTA (CK) promoter crossed to tetO-p25 mice (005706; JAX) in the absence of doxycycline, with CK mice as littermate controls.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Cruz et al., 2003 source

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

    Interpretationreasoning

    The lack of peripheral effects could potentially be explained by A11's preferential brain accumulation, absence from the blood, and rapid overall clearance from the body (half-life <4 h).

    “This could potentially be explained by its preferential brain accumulation, absence from the blood, and rapid overall clearance from the body (half-life of <4 h).”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    All replicate ChIP-Seq data sets passed QC based on ENCODE ChIP-Seq data standards.

    “All replicate data sets passed QC based on ENCODE ChIP-Seq data standards based on read quality, read mapping statistics, library complexity, and strand cross-correlation analysis to measure signal-to-noise ratios.”
    Quote in the paperNot a substantial claim

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

    Interpretationnone given

    For experiments with 6-wk induced CK-p25 mice CK littermates, only females were used.

    “For experiments with 6-wk induced CK-p25 mice CK littermates, only females were used.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    A11 is a first-in-class molecule that converts PU.1 from a transcriptional activator to a transcriptional repressor, resulting in a controlled state of microglial inflammation.

    “A11 represents a first-in-class molecule of drugs that converts PU.1 from a transcriptional activator to a transcriptional repressor, resulting in a controlled state of microglial inflammation.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). The quote only partly states this claim.

    Study design and conductown data

    Results in Figure S5 panels were repeated in two separate batches of experiments.

    “Results in this panel were repeated in two separate batches of experiments.”
    Quote in the paperNot a substantial claim

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

    Interpretationreasoning

    The result of microglial response to neurodegeneration may be beneficial, neutral, or detrimental depending on the type of disease and its point of progression.

    “The complex nature of microglial activation suggests that the result of microglial response to neurodegeneration may be beneficial, neutral, or detrimental depending on the type of disease and its point of progression.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    The mouse models used are imperfect models of AD-related neurodegeneration.

    “While these results suggest considerable translational potential for A11, we acknowledge that these are imperfect models of AD-related neurodegeneration.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: Currently, no mouse strain exhibits all AD hallmarks at once.
    • Rests on an assumption: The CK-p25 mouse model exhibits rapid and inducible neurodegeneration and neuroinflammation, whereas P301STau and 5XFAD mice reproduce tau and amyloid-β accumulation, respectively.

  • May combine two claims.

    Interpretationown data

    Taken together, these findings show that A11 preferentially distributes to the brain following systemic injection and that its administration reduces neuropathology, including tau pathology and β-amyloid accumulation, and increases cognitive performance in several different mouse models of AD without observable side effects.

    “Taken together, these findings show that A11 preferentially distributes to the brain following systemic injection and that its administration reduces neuropathology, including tau pathology and β-amyloid accumulation, and increases cognitive performance in several different mouse models of AD without observable side effects.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    A11 treatment can alleviate working memory deficits in AD mouse models without observable side effects.

    “This suggested that A11 treatment can alleviate working memory deficits in AD mouse models without observable side effects.”
    Quote in the paperSubstantial claim

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

    Study design and conductown data

    Results in Figure 3 panel e were repeated in three separate batches of experiments.

    “Results in this panel were repeated in three separate batches of experiments.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based). The quote only partly states this claim.

    Study design and conductnone given

    Results in Figure S4 panels were repeated in two separate batches of experiments.

    “Results in this panel were repeated in two separate batches of experiments.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Images were adjusted for brightness and contrast equally for the whole image for both control and experimental samples.

    “Images were adjusted for brightness and contrast equally for the whole image for both the control and experimental samples.”
    Quote in the paperNot a substantial claim
71 of 71

  • The paper relies on this without support.

    iMGLs derived from iPSCs or embryonic stem cells are a valid model of human microglia for assessing PU.1 inhibitor effects.

    “We tested the inhibitory effects of the top six hits on phagocytic activity and inflammatory response in human microglia-like cells derived from either iPSCs or embryonic stem cells (ES iMGLs; Fig. S2, d and e).”

  • The paper relies on this without support.

    Preclinical efficacy of A11 in mouse models and iMGLs predicts clinical application for Alzheimer's disease.

    “We discovered a new class of molecules with promising potential for clinical application that reduce hallmarks of neuroinflammation by inhibiting the PU.1 transcription factor, a master regulator of inflammation.”

  • The paper relies on this without support.

    Currently, no mouse strain exhibits all AD hallmarks at once.

    “Currently, strains that exhibit all of these hallmarks at once do not exist.”

  • The paper relies on this without support.

    The CK-p25 mouse model exhibits rapid and inducible neurodegeneration and neuroinflammation, whereas P301STau and 5XFAD mice reproduce tau and amyloid-β accumulation, respectively.

    “The CK-p25 mouse model exhibits rapid and inducible neurodegeneration and neuroinflammation, whereas the P301STau and 5XFAD mice reproduce tau and amyloid-β accumulation, respectively.”

  • The paper relies on this without support.

    Targeting PU.1 therapeutically carries potential for alterations in hematopoiesis.

    “An important consideration of any therapeutic targeting PU.1 is the potential for alterations in hematopoiesis.”
5 of 5

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?

  • Conditional inactivation or pharmacological inhibition of PU.1 in an Alzheimer's disease mouse model ameliorated neuroinflammation, prevented microgliosis, and improved cognitive performance.

    “A recent study showed that conditional inactivation or pharmacological inhibition of PU.1 in AD mouse model ameliorated neuroinflammation, prevented microgliosis and improved cognitive performance (Ralvenius, Mungenast et al., 2023).”

    From Pu.1/Spi1 dosage controls the turnover and maintenance of microglia in zebrafish and mammals (2023) · cites this paper as Ralvenius, Mungenast et al., 2023

    The Results (‘Validating PU.1 as an AD drug target’; page not supplied) report that conditional PU.1 ablation reduced microgliosis by 33%, but do not report a cognitive benefit from ablation. The Discussion reports that pharmacological inhibition with A11 reduced microgliosis and mitigated memory deficits. The paper supports reduced—not prevented—microgliosis, and attributes improved cognitive performance to A11 treatment, not to both interventions. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • Conditional inactivation or pharmacological inhibition of PU.1 in the AD mouse model ameliorated neuroinflammation, prevented microgliosis, and improved cognitive performance.

    “conditional inactivation or pharmacological inhibition of PU.1 in the AD mouse model ameliorated neuroinflammation, prevented microgliosis, and improved cognitive performance (Ralvenius et al., 2023).”

    From Pu.1/Spi1 dosage controls the turnover and maintenance of microglia in zebrafish and mammals (2025) · cites this paper as Ralvenius et al., 2023

    The Results section, “Validating PU.1 as an AD drug target” (page unavailable), reports that conditional PU.1 ablation reduced microgliosis in CK-p25 mice, but does not report a cognitive test for those mice. The Results section on A11 and the Discussion (pages unavailable) report reduced microgliosis and improved memory-task performance after pharmacological inhibition. Thus, improved cognitive performance is supported for A11 treatment, not for conditional inactivation; the paper also reports reduced, rather than wholly prevented, microgliosis. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • Conditional inactivation of PU.1 by feeding tamoxifen-containing diet in Alzheimer's disease model CK-p25 mice led to a 33% reduction in microglia number at their young ages.

    “A recent work found that conditional inactivation of PU.1 by feeding the animals with tamoxifen-containing diet in the Alzheimer’s disease model CK-p25 mice led to a 33% reduction in microglia number at their young ages.”

    From Pu.1/Spi1 dosage controls the turnover and maintenance of microglia in zebrafish and mammals (2025) · cites this paper as Ralvenius et al., 2023

    The Results section (“Validating PU.1 as an AD drug target”; page unavailable) reports that conditional PU.1 ablation reduced hippocampal microgliosis by 33% in male CK-p25 mice after six weeks of p25 induction. Materials and methods (“Mouse experiments”; page unavailable) confirms tamoxifen-containing chow, but the reported 33% outcome is microgliosis, not explicitly microglial cell number, and the paper does not describe those mice as young. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • Oral administration of a functional PU.1 inhibitor for six weeks reduced amyloid plaque deposition in a 1-year-old amyloid mouse model and decreased tau phosphorylation in a 1-year-old tauopathy mouse model.

    “In contrast, another recent study reported that oral administration of a functional PU.1 inhibitor for six weeks reduceed amyloid plaque deposition in a 1‐year‐old amyloid mouse model and decreases tau phosphorylation in a 1‐year‐old tauopathy mouse model [42].”

    From Targeting Microglial Transcriptional Reprogramming as a Therapy Strategy for Alzheimer's Disease (2026) · cites this paper as [42]

    The Results section reports that six weeks of A11 treatment reduced phosphorylated tau in 1-year-old P301STau mice and hippocampal β-amyloid accumulation in 1-year-old 5XFAD mice. However, A11 was administered by daily intraperitoneal injection, not orally; the claim's administration route is unsupported. (LLM, conf=0.94)

    Experimental — accuracy not yet measured

  • In later stages of Alzheimer's disease, reducing PU.1 activity while increasing MEF2C activity inhibits harmful inflammation and supports neuroprotective microglial functions.

    “in the later stages, it might be beneficial to reduce PU.1 activity while increasing MEF2C activity to inhibit harmful inflammation and support neuroprotective microglial functions”

    From Targeting Microglial Transcriptional Reprogramming as a Therapy Strategy for Alzheimer's Disease (2026) · cites this paper as [42]

    The Results section reports that reducing PU.1 activity decreased microgliosis and neurodegeneration in an AD-related mouse model, supporting the PU.1 part of the proposal. The paper does not establish that this approach is specifically beneficial in later-stage Alzheimer’s disease, and a search for MEF2C found no support for increasing its activity or for the proposed combined effect. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

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.

  • Conflicting units for apparent permeability

    Minor

    The Results give Papp in inverse-centimeters times inverse-seconds, while the permeability method defines it in centimeters per second.

  • Conflicting sample sizes for Figure 1c

    Minor

    The Figure 1c caption gives three mice for the PU.1WT/WT group, then says there were four for all conditions.

  • Conflicting blood collection methods

    Minor

    The Figure S5 caption says blood for panel d was obtained by decapitation, whereas the corresponding mouse-blood flow-cytometry method says it was collected by cardiac puncture.

3 of 3

Cited works that were retracted

Works this paper cites that were later retracted, from the retraction notices publishers deposit. Major when the notice came out before this paper was published.

  • Cites a retracted work: 10.1212/wnl.0000000000200176

    Info

    The cited work was retracted on 2025-04-14 (notice 10.1212/wnl.0000000000213572), not before this paper was published (2023-08-29).

    Cited work 10.1212/wnl.0000000000200176 · retracted 2025-04-14 · notice 10.1212/wnl.0000000000213572 · this paper published 2023-08-29

1 of 1

Cites

1 retracted

Flagged first, then by citations

4 of the 83 references couldn’t be matched to a known paper.

Research works printed but not linked: 1

  1. Alzheimer, 1907, Über eine eigenartige Erkrankung der Hirnrinde, Allg. Zeitschr. Psychiatr., 64, 146

Cited by

Sorted by citations

Related

Authors
Ralvenius, W. T., Mungenast, A. E., Woolf, H., Huston, M. M., Gillingham, T. Z., Godin, S. K., Penney, J., Cam, H. P., Gao, F., Fernandez, C. G., Czako, B., Lightfoot, Y., Ray, W. J., Beckmann, A., Goate, A. M., Marcora, E., Romero-Molina, C., Ayata, P., Schaefer, A., Gjoneska, E., & Tsai, L.-H.
Affiliations
  • Massachusetts Institute of Technology (US)
  • The University of Texas MD Anderson Cancer Center (US)
  • The University of Texas MD Anderson Cancer Center 3 The Neurodegeneration Consortium, Therapeutics Discovery Division, , Houston, TX, USA
  • Icahn School of Medicine at Mount Sinai (US)
  • Friedman Brain Institute, Icahn School of Medicine at Mount Sinai 4 Nash Family Department of Neuroscience, , New York, NY, USA
  • The Graduate Center, CUNY (US)
  • Max Planck Institute for Biology of Ageing (DE)
  • National Institutes of Health (US)
  • Broad Institute (US)
Corresponding author
Gjoneska, Elizabeta
ORCID
Venue
Journal of Experimental Medicine
Publisher
Rockefeller University Press
Published
2023-08-29
Volume
220
Issue
11
ISSN
  • 0022-1007 (ISSN-L)
  • 1540-9538
DOI
10.1084/jem.20222105
PMC
PMC10465325
OpenAlex
W4386246035
Semantic Scholar
9a1eb3e6eddd7081445ea4a32d655fbe41117814
Cited by
31
References
83
Citations by year
2026: 7 · 2025: 15 · 2024: 7 · 2023: 2
FWCI
2.61
Type
journal article
Language
en
Open access
Licence
Locations
  • The Journal of Experimental Medicine (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
  • IslandScholar (University of Prince Edward Island) (submittedVersion, cc-by-nc-sa)
  • PubMed Central (submittedVersion, cc-by-nc-sa)
  • DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc-sa)
Funders
  • National Institutes of Health (1U01AG066757-01)
  • JPB Foundation
  • Swiss National Science Foundation (177920, 171909)
  • Alzheimer’s Association (AARF-19-616816)
  • University of Texas MD Anderson Cancer Center
  • National Cancer Institute (CA016672)
  • Cancer Prevention and Research Institute of Texas (RP120348, RP170002)
  • Robert A. and Renee E. Belfer Family Foundation
  • Picower Institute for Learning and Memory
  • Halis Family Foundation
Topics
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
  • Life Sciences › Neuroscience › Neurology › Neurological Disease Mechanisms and Treatments
Keywords
neuroinflammation, Alzheimer's disease, PU.1, transcription factor binding motifs, microglia, HDAC1, MeCP2, SIN3A
MeSH terms
Neuroinflammatory Diseases, Alzheimer Disease

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 2026-10-07.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-07.

Cited sources are checked by gpt-6-sol.

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 ›

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