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Cited by 4Alzheimer's disease research and treatmentsHistone Deacetylase Inhibitors ResearchWnt/β-catenin signaling in development and cancer

CI-994 is a dual modulator of class I HDACs and Wnt/β-catenin signaling for the treatment of Alzheimer's disease

Open access
Lu, Wenyan, Kawatani, Keiji, Ren, Yingxue, Nambara, Toshihiko, Jia, Lin, Jeevaratnam, Suren, Lee, Eunmi, Martinez, Paula Rodriguez +7 more·2026·Alzheimer's Research & Therapy
DOI 10.1186/s13195-026-01982-0PMC PMC12977471
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Abstract

BACKGROUND: Growing evidence supports that epigenetic dysregulation through histone deacetylases (HDACs) plays a critical role in synaptic dysfunction and memory loss in Alzheimer’s disease (AD), and that HDACs have been highlighted as an attractive class of targets for AD therapy. Moreover, restoring Wnt/β-catenin signaling, which is greatly suppressed in AD brains, is a promising therapeutic strategy. CI-994 is an orally active class I HDAC inhibitor that has undergone several phase II/III clinical trials on cancer treatment. Importantly, CI-994 can cross the blood–brain barrier and is a cognitive enhancer. METHODS: Wnt activity was initially examined by Wnt reporter activity assay in Wnt3A-expression HEK293 cells, and profiling HDAC inhibition was performed against 10 individual HDACs. Activities of CI-994 on class I HDACs and Wnt/β-catenin signaling were further tested in HEK293 cells, LRP6-expressing HT1080 cells and neuronal SH-SY5Y cells. The therapeutic effects of CI-994 were examined in patient-specific iPSC-derived neurons and cerebral organoids carrying APOE ε4/ε4 genotype or MAPT p.P301L mutation. RESULTS: We herein report that CI-994 is not only a potent class I HDAC inhibitor but also an activator of Wnt/β-catenin signaling. Mechanistically, activation of Wnt/β-catenin signaling by CI-994 is associated with stabilizing Wnt co-receptor LRP6 protein and modulating HDAC activity. Importantly, CI-994 significantly increases histone acetylation, activates Wnt/β-catenin signaling, and decreases tau phosphorylation in patient-specific iPSC-derived cerebral organoids carrying APOE ε4/ε4 genotype or MAPT p.P301L mutation. Moreover, studies with the specific Wnt inhibitor LGK974 demonstrate that activation of Wnt/β-catenin signaling contributes to CI-994-induced the inhibition of tau phosphorylation in the iPSC-derived cerebral organoids. Additionally, CI-994 increases synaptic protein levels, enhances spontaneous synaptic firing and network formation and decreases tau phosphorylation in iPSC-derived neurons. Finally, RNA sequencing, combined with RT-qPCR validation, of the iPSC-derived cerebral organoids reveals that CI-994 significantly regulates genes associated with synapse plasticity and cognitive function including NEUROD1, CACNA1G, NRGN, NRTN, SLC7A10 and OMG. CONCLUSION: Our findings suggest that CI-994 can be repurposed as a novel therapeutic agent for AD therapy.

AI snapshot

Written from the paper’s full text.

TL;DR

CI-994 is reported as a dual modulator that inhibits class I HDACs and activates Wnt/β-catenin signaling, partly by stabilizing LRP6. In patient-specific iPSC-derived neurons and cerebral organoids, it increases histone acetylation and synaptic markers, reduces tau phosphorylation, and modulates synapse/cognition-related genes.

Key findings

  • CI-994 activated Wnt reporter activity in Wnt3A-expressing HEK293 cells with an EC50 of ~5.8 µM and inhibited HDAC1, HDAC2, and HDAC3 with IC50 values of 617 nM, 593 nM, and 472 nM, respectively; at 10 µM it did not substantially inhibit other tested HDACs.
  • In iPSC-derived cerebral organoids carrying APOE ε4/ε4 or MAPT p.P301L, CI-994 at 4 and 8 µM increased histone H3 acetylation and β-catenin and decreased tau phosphorylation detected by AT8 and AT180.
  • The Wnt inhibitor LGK974 reduced CI-994-induced tau phosphorylation inhibition: CI-994 alone produced an 80% reduction, while CI-994 plus LGK974 produced a 67% reduction versus vehicle in APOE ε4/ε4 organoids.
  • RNA-seq of CI-994-treated APOE ε4/ε4 organoids identified 835 differentially expressed genes, including 492 upregulated; NRGN, NRTN, and SLC7A10 were among the top 10 upregulated genes, and OMG was among the top 10 downregulated genes. CI-994 also increased spontaneous spike frequency and burst firing and synaptic proteins in iPSC-derived neurons.

Methods

  • In vitro assays included Wnt reporter activity in Wnt3A-expressing HEK293 cells, HDAC inhibition profiling against 10 HDACs, IC50 determination for HDAC1–3, and Western blotting in HEK293, LRP6-expressing HT1080, and SH-SY5Y cells.
  • Patient-specific iPSC lines were from an APOE ε4/ε4 AD patient and a MAPT p.P301L carrier; iPSC-derived cerebral organoids were treated with CI-994 at 0, 4, and 8 µM for 3 days, with three organoids pooled per sample and three samples per group.
  • iPSC-derived neurons were analyzed by microelectrode array for spontaneous spikes and burst firing over 8 weeks with CI-994 at 4 µM, and cerebral organoids were analyzed by RNA-seq, RT-qPCR, and Western blotting.

Limitations

  • The paper states that the mechanism by which CI-994 stabilizes LRP6 is unclear and requires further investigation.
  • The paper states that it is unclear whether HDAC inhibition and Wnt/β-catenin activation contribute equally to in vivo therapeutic effects; LGK974 studies suggest tau phosphorylation inhibition is mainly associated with class I HDAC inhibition, and HDAC IC50 values are much lower than the Wnt activation EC50.
  • Evident from the text: the therapeutic evidence is from cell and patient-specific iPSC-derived models, not in vivo AD efficacy studies; some organoid comparisons, such as β-catenin with LGK974, lacked statistical significance due to sample variance.

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

Integrity signals

Registry

Crossref · 10.1186/s13195-026-01982-0

Cites

82 of 82 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

89 of 125 cited sources checked · 42 supported · 2 flagged · accuracy not yet measured

  • Premisecitation

    Inhibition of class I HDACs is associated with activation of Wnt/β-catenin signaling.

    “It has been demonstrated that inhibition of class I HDACs is associated with activation of Wnt/β-catenin signaling [42–47].”
    Quote in the paperSubstantial claim4 of 6 sources checked
    • cites [42] Ye F, Chen Y, Hoang T, Montgomery RL, Zhao XH, Bu H, et al. HDAC1 and HDAC2 regulate oligodendrocyte differentiation by disrupting the beta-catenin-TCF interaction. Nat Neurosci. 2009;12(7):829–38. source
    • cites [43] Wu Z, Wei D, Gao W, Xu Y, Hu Z, Ma Z, et al. TPO-induced metabolic reprogramming drives liver metastasis of colorectal cancer CD110+ tumor-initiating cells. Cell Stem Cell. 2015;17(1):47–59. source
    • cites [44] Masoumi KC, Daams R, Sime W, Siino V, Ke H, Levander F, et al. NLK-mediated phosphorylation of HDAC1 negatively regulates Wnt signaling. Mol Biol Cell. 2017;28(2):346–55. source

      The Results section reports that active HDAC1 represses a Wnt/β-catenin-responsive Tcf/Lef reporter, while a catalytically inactive HDAC1 mutant does not. The Discussion identifies HDAC1 as a class I HDAC. The paper therefore supports a role for HDAC1 activity in limiting this signaling, but does not demonstrate that inhibition of class I HDACs as a group activates it. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [45] You H, Li Q, Kong D, Liu X, Kong F, Zheng K, et al. The interaction of canonical Wnt/beta-catenin signaling with protein lysine acetylation. Cell Mol Biol Lett. 2022;27(1):7. source

      The section “Targeting protein acetylation to suppress Wnt/β-catenin signaling” (page unavailable) says that mocetinostat, an inhibitor of class I and IV HDACs, suppresses Wnt/β-catenin signaling. Although the paper also describes HDAC1 and HDAC2 as repressors of Wnt-dependent transcription, its stated effect of an HDAC inhibitor is contrary to the claimed activation. (LLM, conf=0.86)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [46] Liu C, Zhang Y, Li J, Wang Y, Ren F, Zhou Y, et al. p15RS/RPRD1A (p15INK4b-related sequence/regulation of nuclear pre-mRNA domain-containing protein 1A) interacts with HDAC2 in inhibition of the Wnt/beta-catenin signaling pathway. J Biol Chem. 2015;290(15):9701–13. source
    • cites [47] Billin AN, Thirlwell H, Ayer DE. Beta-catenin-histone deacetylase interactions regulate the transition of LEF1 from a transcriptional repressor to an activator. Mol Cell Biol. 2000;20(18):6882–90. source

      The Results report that the HDAC inhibitor TSA stimulated a LEF/TCF-dependent reporter and permitted β-catenin-dependent activation. But TSA alone did not activate the siamois promoter; the paper distinguishes relief of HDAC1-mediated repression from activation, which also required LEF1 and β-catenin. Thus the claim overgeneralizes a conditional reporter finding to activation of Wnt/β-catenin signaling. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
  • Premisecitation

    Epigenetic dysregulation through HDACs plays a critical role in synaptic dysfunction and memory loss in Alzheimer's disease.

    “Growing evidence supports that epigenetic dysregulation through histone deacetylases (HDACs) plays a critical role in synaptic dysfunction and memory loss in Alzheimer’s disease”
    • Also stated in: § Introduction · plain
      “Histone deacetylases (HDACs) play important roles in synaptic plasticity and cognitive function through the epigenetic regulation of gene expression in the brain [4–8].”
    Quote in the paperSubstantial claim4 of 5 sources checked
    • cites [4] Falkenberg KJ, Johnstone RW. Histone deacetylases and their inhibitors in cancer, neurological diseases and immune disorders. Nat Rev Drug Discov. 2014;13(9):673–91. source
    • cites [5] Burns AM, Graff J. Cognitive epigenetic priming: leveraging histone acetylation for memory amelioration. Curr Opin Neurobiol. 2021;67:75–84. source

      The sections “HDAC inhibition increases histone acetylation” and “HDAC inhibition enhances transcription” report memory impairment, altered histone acetylation, and misregulated synaptic-plasticity genes in preclinical Alzheimer’s disease models; HDAC inhibitors reversed some of these changes. The paper therefore supports a role for HDAC-related dysregulation, but the claim’s unqualified “critical role” in Alzheimer’s disease extends beyond the model-based evidence presented. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [6] Pao PC, Tsai LH. Histone deacetylases 1 and 2 in memory function. ACS Chem Neurosci. 2022;13(7):848–58. source
    • cites [7] Kumar V, Kundu S, Singh A, Singh S. Understanding the role of histone deacetylase and their inhibitors in neurodegenerative disorders: current targets and future perspective. Curr Neuropharmacol. 2022;20(1):158–78. source
    • cites [8] Kabir F, Atkinson R, Cook AL, Phipps AJ, King AE. The role of altered protein acetylation in neurodegenerative disease. Front Aging Neurosci. 2022;14:1025473. source

      The HDAC enzymes section (page unavailable) says HDAC2 regulates memory formation and synaptic plasticity, and its table lists several HDACs as involved in Alzheimer’s disease. The altered microtubule acetylation section (page unavailable) also links HDAC6 activity to impaired cognition in AD mouse models. These passages do not establish the more specific claim that epigenetic dysregulation through HDACs plays a critical role in both synaptic dysfunction and memory loss in Alzheimer’s disease. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
  • Premisecitation

    CI-994 can cross the blood-brain barrier.

    “CI-994 can cross the blood–brain barrier and is a cognitive enhancer.”
    • Also stated in: § Introduction · plain
      “which can across the blood–brain barrier [16, 17].”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [16] Riva L, Blaney SM, Dauser R, Nuchtern JG, Durfee J, McGuffey L, et al. Pharmacokinetics and cerebrospinal fluid penetration of CI-994 (N-acetyldinaline) in the nonhuman primate. Clin Cancer Res. 2000;6(3):994–7.

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

    • cites [17] Graff J, Joseph NF, Horn ME, Samiei A, Meng J, Seo J, et al. Epigenetic priming of memory updating during reconsolidation to attenuate remote fear memories. Cell. 2014;156(1–2):261–76. source
  • Premisecitation

    Wnt/β-catenin signaling is greatly suppressed in AD brains.

    “restoring Wnt/β-catenin signaling, which is greatly suppressed in AD brains, is a promising therapeutic strategy”
    • Also stated in: § Introduction · hedged
      “Increasing evidence indicates that Wnt/β-catenin signaling is diminished by multiple pathogenic pathways in AD brain, and that restoring Wnt/β-catenin signaling represents a unique opportunity for the rational design of novel AD therapies [14].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [14] Jia L, Pina-Crespo J, Li Y. Restoring Wnt/beta-catenin signaling is a promising therapeutic strategy for Alzheimer’s disease. Mol Brain. 2019;12(1):104. source
  • Premisecitation

    ITSA-1 counteracts the activity of TSA, a potent inhibitor of class I and II HDACs, on histone acetylation and transcriptional activation.

    “ITSA-1 is an activator of HDACs and counteracts the activity of TSA, a potent inhibitor of class I and II HDACs, on histone acetylation and transcriptional activation [48–50].”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [48] Koeller KM, Haggarty SJ, Perkins BD, Leykin I, Wong JC, Kao MC, et al. Chemical genetic modifier screens: small molecule trichostatin suppressors as probes of intracellular histone and tubulin acetylation. Chem Biol. 2003;10(5):397–410. source
    • cites [49] Su Q, Li T, He PF, Lu XC, Yu Q, Gao QC, et al. Trichostatin A ameliorates Alzheimer’s disease-related pathology and cognitive deficits by increasing albumin expression and Abeta clearance in APP/PS1 mice. Alzheimers Res Ther. 2021;13(1):7. source

      The Results report that ITSA-1 suppressed TSA-induced histone H4 acetylation in BV2 cells and reversed TSA-induced albumin expression in endothelial cells. The Background identifies TSA as an HDAC inhibitor. Our copy does not establish that ITSA-1 activates HDACs, that TSA specifically inhibits class I and II HDACs, or that ITSA-1 counteracts transcriptional activation generally. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [50] Behera J, Kelly KE, Voor MJ, Metreveli N, Tyagi SC, Tyagi N. Hydrogen sulfide promotes bone homeostasis by balancing inflammatory cytokine signaling in CBS-deficient mice through an epigenetic mechanism. Sci Rep. 2018;8(1):15226. source

      The Results section (page unavailable) calls ITSA-1 “Inhibitor-1 of Trichostatin A” and reports that it reversed changes in HDAC activity and histone acetylation in CBS-deficient mice. The Discussion (page unavailable) calls it an HDAC activator and reports suppressed inflammatory cytokine expression. Our copy does not show ITSA-1 directly counteracting TSA’s effects on histone acetylation or transcriptional activation, or describe TSA as a potent inhibitor of class I and II HDACs. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
  • May combine two claims. 24
  • Classified as probably not a substantial claim. 24
  • The quote only partly states this claim. 3

  • May combine two claims.

    Premisecitation

    CI-994 is a molecular memory aid and promotes memory formation and cognitive function in vivo.

    “CI-994 is a molecular memory aid and promotes memory formation and cognitive function in vivo [9, 17, 21–24].”
    Quote in the paperNot a substantial claim6 of 6 sources checked
    • cites [9] Marinho D, Ferreira IL, Lorenzoni R, Cardoso SM, Santana I, Rego AC. Reduction of class I histone deacetylases ameliorates ER-mitochondria cross-talk in Alzheimer’s disease. Aging Cell. 2023;22(8):e13895. source

      The Introduction (page unavailable) identifies tacedinaline as CI-994 and reports improved cognitive and memory dysfunction in two mouse models, supporting an in vivo memory and cognition benefit. It does not establish that CI-994 promotes memory formation specifically; the paper’s own in vivo results measure hippocampal pathology and molecular changes, not memory formation. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [17] Graff J, Joseph NF, Horn ME, Samiei A, Meng J, Seo J, et al. Epigenetic priming of memory updating during reconsolidation to attenuate remote fear memories. Cell. 2014;156(1–2):261–76. source

      The Results section, “By Using HDAC2-Targeting HDAC Inhibitors, Remote Fear Memories Can Be Attenuated” (page unavailable), reports that CI-994 combined with recall and extinction training produced persistent extinction of remote fear memories in mice. This supports a role in memory updating in vivo, but not the broader claim that CI-994 promotes memory formation and cognitive function generally; the Results also state that the inhibitor alone had no behavioral effect. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [21] Zhang S, Fujita Y, Matsuzaki R, Yamashita T. Class I histone deacetylase (HDAC) inhibitor CI-994 promotes functional recovery following spinal cord injury. Cell Death Dis. 2018;9(5):460. source

      The Introduction says previous evidence suggests CI-994 attenuates fear memories, rather than promoting memory formation. The Discussion reports improved motor recovery after spinal cord injury, not improved cognitive function. (LLM, conf=0.76)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [22] Cooper A, Butto T, Hammer N, Jagannath S, Fend-Guella DL, Akhtar J, et al. Inhibition of histone deacetylation rescues phenotype in a mouse model of Birk-Barel intellectual disability syndrome. Nat Commun. 2020;11(1):480. source

      The Results section, “CI-994 rescues the behavioral phenotype of Kcnk9KOmat animals” (page not provided), reports improved Y-maze performance, a measure of working memory, in a mouse model of Birk-Barel syndrome. It reports no significant working-memory improvement in wild-type mice. The study therefore supports an in vivo rescue of impaired working memory, but not the broader claim that CI-994 generally promotes memory formation or cognitive function. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [23] McClarty B, Rodriguez G, Dong H. Dose effects of histone deacetylase inhibitor tacedinaline (CI-994) on antipsychotic haloperidol-induced motor and memory side effects in aged mice. Front Neurosci. 2021;15:674745. source

      The Results report that CI-994 improved recognition memory in haloperidol-treated aged mice, supporting an in vivo memory benefit. But the study did not test memory formation or broad cognitive function, and Figure 2 reports no significant memory difference between CI-994 alone and vehicle in aged mice. The claim generalizes beyond the effect this study demonstrated. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [24] Burns AM, Farinelli-Scharly M, Hugues-Ascery S, Sanchez-Mut JV, Santoni G, Graff J. The HDAC inhibitor CI-994 acts as a molecular memory aid by facilitating synaptic and intracellular communication after learning. Proc Natl Acad Sci U S A. 2022;119(22):e2116797119. source

  • May combine two claims.

    Premisecitation

    CI-994 is an orally active class I HDAC inhibitor.

    “CI-994 is an orally active class I HDAC inhibitor that has undergone several phase II/III clinical trials on cancer treatment.”
    • Also stated in: § Introduction · plain
      “The benzamide CI-994 is an orally bioavailable class I HDAC inhibitor which can across the blood–brain barrier [16, 17].”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [16] Riva L, Blaney SM, Dauser R, Nuchtern JG, Durfee J, McGuffey L, et al. Pharmacokinetics and cerebrospinal fluid penetration of CI-994 (N-acetyldinaline) in the nonhuman primate. Clin Cancer Res. 2000;6(3):994–7.
    • cites [17] Graff J, Joseph NF, Horn ME, Samiei A, Meng J, Seo J, et al. Epigenetic priming of memory updating during reconsolidation to attenuate remote fear memories. Cell. 2014;156(1–2):261–76. source

      The Results section, “By Using HDAC2-Targeting HDAC Inhibitors,” identifies CI-994 as a selective class I HDAC inhibitor. Our copy does not establish that it is orally active; the Results and Experimental Procedures describe intraperitoneal administration instead. (LLM, conf=0.87)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    CI-994 is a cognitive enhancer / improves cognitive function.

    “CI-994 can cross the blood–brain barrier and is a cognitive enhancer.”
    • Also stated in: § Introduction · hedged
      “Mounting evidence indicates that CI-994 acts as a molecular memory aid and improves cognitive function in several mouse models of neurodegenerative disorders [9, 17, 21–24].”
    Quote in the paperSubstantial claim6 of 6 sources checked
    • cites [9] Marinho D, Ferreira IL, Lorenzoni R, Cardoso SM, Santana I, Rego AC. Reduction of class I histone deacetylases ameliorates ER-mitochondria cross-talk in Alzheimer’s disease. Aging Cell. 2023;22(8):e13895. source
    • cites [17] Graff J, Joseph NF, Horn ME, Samiei A, Meng J, Seo J, et al. Epigenetic priming of memory updating during reconsolidation to attenuate remote fear memories. Cell. 2014;156(1–2):261–76. source

      The Results section ('By Using HDAC2-Targeting HDAC Inhibitors, Remote Fear Memories Can Be Attenuated'; page unavailable) reports sustained CI-994 levels in the brain after systemic administration, supporting brain penetration. It also reports improved extinction of remote fear memories when CI-994 was paired with memory recall and extinction training, but does not establish that CI-994 generally improves cognitive function or acts as a cognitive enhancer on its own. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [21] Zhang S, Fujita Y, Matsuzaki R, Yamashita T. Class I histone deacetylase (HDAC) inhibitor CI-994 promotes functional recovery following spinal cord injury. Cell Death Dis. 2018;9(5):460. source

      The Introduction says CI-994 has been reported to attenuate fear memories, while the Results and Discussion report motor recovery after spinal cord injury—not improved cognitive function. A search for cognition and cognitive enhancement found no passage in our copy establishing that CI-994 is a cognitive enhancer. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [22] Cooper A, Butto T, Hammer N, Jagannath S, Fend-Guella DL, Akhtar J, et al. Inhibition of histone deacetylation rescues phenotype in a mouse model of Birk-Barel intellectual disability syndrome. Nat Commun. 2020;11(1):480. source

      The Results section ('CI-994 rescues the behavioral phenotype of Kcnk9KOmat animals'; page unavailable) reports improved Y-maze working-memory performance in a mouse model after CI-994 treatment. The same section reports no significant Y-maze improvement in wild-type mice, so the unqualified description of CI-994 as a cognitive enhancer is broader than the evidence. The paper also reports that administered CI-994 reaches the brain, consistent with blood–brain barrier passage. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [23] McClarty B, Rodriguez G, Dong H. Dose effects of histone deacetylase inhibitor tacedinaline (CI-994) on antipsychotic haloperidol-induced motor and memory side effects in aged mice. Front Neurosci. 2021;15:674745. source

      The Results (Dose Dependent Effects of CI-994 on Haloperidol Induced Memory Impairment; page unavailable) show that CI-994 improved recognition memory in aged mice whose memory was impaired by haloperidol. The same section reports no significant memory difference between CI-994 alone and vehicle, so the unqualified description of CI-994 as a cognitive enhancer overstates this study’s finding. Our copy also does not establish the citing sentence’s blood–brain-barrier claim (searched: blood–brain barrier, cross). (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [24] Burns AM, Farinelli-Scharly M, Hugues-Ascery S, Sanchez-Mut JV, Santoni G, Graff J. The HDAC inhibitor CI-994 acts as a molecular memory aid by facilitating synaptic and intracellular communication after learning. Proc Natl Acad Sci U S A. 2022;119(22):e2116797119. source

      The Results section reports that CI-994 improved memory retention in mice when paired with contextual fear conditioning, supporting its description as a cognitive enhancer in that setting. The Discussion reports brain-region-specific effects after systemic administration, but our copy does not directly establish that CI-994 crosses the blood–brain barrier or that it improves cognitive function more generally. (LLM, conf=0.82)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Wnt/β-catenin signaling is crucial for neuronal survival and neurogenesis and plays important roles in regulating synaptic plasticity and blood-brain barrier integrity and function.

    “Wnt/β-catenin signaling is not only crucial for neuronal survival and neurogenesis but also plays important roles in regulating synaptic plasticity and blood–brain barrier integrity and function [14].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [14] Jia L, Pina-Crespo J, Li Y. Restoring Wnt/beta-catenin signaling is a promising therapeutic strategy for Alzheimer’s disease. Mol Brain. 2019;12(1):104. source

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

    Premisecitation

    CI-994 has undergone phase II clinical trials for advanced pancreatic cancer, nonsmall cell lung cancer and multiple myeloma, and phase III clinical trials in combination with gemcitabine for advanced nonsmall-cell lung and pancreatic cancers (discontinued).

    “CI-994 has undergone phase II clinical trials for advanced pancreatic cancer, nonsmall cell lung cancer and multiple myeloma [18–20], and has been in phase III clinical trials in combination with gemcitabine for advanced nonsmall-cell lung and pancreatic cancers, though this trial was discontinued [20].”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [18] Richards DA, Boehm KA, Waterhouse DM, Wagener DJ, Krishnamurthi SS, Rosemurgy A, et al. Gemcitabine plus CI-994 offers no advantage over gemcitabine alone in the treatment of patients with advanced pancreatic cancer: results of a phase II randomized, double-blind, placebo-controlled, multicenter study. Ann Oncol. 2006;17(7):1096–102. source

      Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured
    • cites [19] Gridelli C, Rossi A, Maione P. The potential role of histone deacetylase inhibitors in the treatment of non-small-cell lung cancer. Crit Rev Oncol Hematol. 2008;68(1):29–36. source
    • cites [20] Avendano C, Menende TC. Chapter 8 - Epigenetic therapy of cancer. Medicinal Chemistry of Anticancer Drugs (Third Edition). 2023. source

  • Classified as probably not a substantial claim.

    Premisecitation

    Hyperphosphorylated forms of tau in neurons is a major hallmark of AD.

    “Hyperphosphorylated forms of the microtubule-associated protein tau in neurons is a major hallmark of AD [2, 3].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [2] Busche MA, Hyman BT. Synergy between amyloid-β and tau in Alzheimer’s disease. Nat Neurosci. 2020;23(10):1183–93. source
    • cites [3] Yamazaki Y, Zhao N, Caulfield TR, Liu CC, Bu G. Apolipoprotein E and Alzheimer disease: pathobiology and targeting strategies. Nat Rev Neurol. 2019;15(9):501–18. source

  • May combine two claims.

    Premisecitation

    Cerebral organoids derived from AD patient-specific iPSCs can recapture patient-specific phenotypes and hence serve as excellent disease models for AD research and drug development.

    “Cerebral organoids derived from AD patient-specific iPSCs can recapture patient-specific phenotypes and hence serve as excellent disease models for AD research and drug development [51–53].”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [51] Sullivan SE, Young-Pearse TL. Induced pluripotent stem cells as a discovery tool for Alzheimer’s disease. Brain Res. 2017;1656:98–106. source

      Section 3.2 reports distinct cellular phenotypes in iPSC-derived neurons from different AD patients, and Section 6.2 discusses their potential for drug screening. But Section 6.3 describes cerebral organoids only as a developing approach that recapitulates aspects of cortical development; it does not establish that AD patient-derived organoids recapture patient-specific disease phenotypes or are already excellent AD drug-development models. Page numbers were not available in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [52] Khakipoor S, Crouch EE, Mayer S. Human organoids to model the developing human neocortex in health and disease. Brain Res. 2020;1742:146803. source

      The Future perspectives section reports that forebrain organoids modeling Alzheimer’s disease mimic amyloid and tau pathology and respond to drug treatment. It does not specify that those organoids came from AD patient-specific iPSCs or recaptured patient-specific phenotypes; it describes organoid-based personalized treatment testing as a future possibility, not a current capability. Page numbers are unavailable in our copy. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [53] Salick MR, Lubeck E, Riesselman A, Kaykas A. The future of cerebral organoids in drug discovery. Semin Cell Dev Biol. 2021;111:67–73. source

  • May combine two claims.

    Premisecitation

    HDAC inhibitors can enhance histone acetylation and ameliorate cognitive impairments in various AD models.

    “HDAC inhibitors can enhance histone acetylation and ameliorate cognitive impairments in various AD models, and HDAC inhibition has been recognized as an attractive therapeutic strategy for AD [4, 5, 7].”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [4] Falkenberg KJ, Johnstone RW. Histone deacetylases and their inhibitors in cancer, neurological diseases and immune disorders. Nat Rev Drug Discov. 2014;13(9):673–91. source
    • cites [5] Burns AM, Graff J. Cognitive epigenetic priming: leveraging histone acetylation for memory amelioration. Curr Opin Neurobiol. 2021;67:75–84. source
    • cites [7] Kumar V, Kundu S, Singh A, Singh S. Understanding the role of histone deacetylase and their inhibitors in neurodegenerative disorders: current targets and future perspective. Curr Neuropharmacol. 2022;20(1):158–78. source

      The Alzheimer’s Disease section (page unavailable) reports that mice treated with a selective HDAC3 inhibitor had increased histone acetylation and cognition; it also reports improved memory in an AD mouse after HDAC2 knockdown. It does not establish that HDAC inhibitors produced both effects across various AD models: the HDAC2 result involves knockdown, not inhibitor treatment. (LLM, conf=0.83)

      Experimental — accuracy not yet measured

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

    Premisecitation

    CI-994 is well tolerated and has undergone phase II/III clinical trials for several types of cancer.

    “Importantly, CI-994 is well tolerated and has undergone phase II/III clinical trials for several types of cancer [18–20].”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [18] Richards DA, Boehm KA, Waterhouse DM, Wagener DJ, Krishnamurthi SS, Rosemurgy A, et al. Gemcitabine plus CI-994 offers no advantage over gemcitabine alone in the treatment of patients with advanced pancreatic cancer: results of a phase II randomized, double-blind, placebo-controlled, multicenter study. Ann Oncol. 2006;17(7):1096–102. source

      The Introduction (page unavailable) says CI-994 was well tolerated in a small pancreatic-cancer trial and describes phase II trials in several cancer types. It does not report a phase III trial of CI-994; the phase III study listed in the References concerns gemcitabine and fluorouracil, not CI-994. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [19] Gridelli C, Rossi A, Maione P. The potential role of histone deacetylase inhibitors in the treatment of non-small-cell lung cancer. Crit Rev Oncol Hematol. 2008;68(1):29–36. source
    • cites [20] Avendano C, Menende TC. Chapter 8 - Epigenetic therapy of cancer. Medicinal Chemistry of Anticancer Drugs (Third Edition). 2023. source

  • May combine two claims.

    Premisecitation

    Depletion of HDAC1/2 stabilizes β-catenin and activates Wnt/β-catenin signaling in oligodendrocyte lineage cells.

    “Firstly, depletion of HDAC1/2 stabilizes β-catenin and activates Wnt/β-catenin signaling in oligodendrocyte lineage cells [42].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [42] Ye F, Chen Y, Hoang T, Montgomery RL, Zhao XH, Bu H, et al. HDAC1 and HDAC2 regulate oligodendrocyte differentiation by disrupting the beta-catenin-TCF interaction. Nat Neurosci. 2009;12(7):829–38. source

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

    Premisecitation

    NRGN is a post-synaptic protein associated with synaptic transmission and memory formation.

    “NRGN is a post-synaptic protein associated with synaptic transmission and memory formation [58].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [58] Hwang H, Szucs MJ, Ding LJ, Allen A, Ren X, Haensgen H, et al. Neurogranin, encoded by the schizophrenia risk gene NRGN, bidirectionally modulates synaptic plasticity via calmodulin-dependent regulation of the neuronal phosphoproteome. Biol Psychiatry. 2021;89(3):256–69. source

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

    Premisecitation

    SLC7A10 (also known as Asc-1) is a sodium-independent neutral amino acid transporter in neurons and astrocytes.

    “SLC7A10 (also known as Asc-1) is a sodium-independent neutral amino acid transporter in neurons and astrocytes [60].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [60] Li Y, Guo Y, Broer A, Dai L, Brӧer S, Yan R. Cryo-EM structure of the human Asc-1 transporter complex. Nat Commun. 2024;15(1):3036. source

  • Classified as probably not a substantial claim.

    Premisecitation

    HDAC inhibition has been recognized as an attractive therapeutic strategy for AD.

    “HDAC inhibitors can enhance histone acetylation and ameliorate cognitive impairments in various AD models, and HDAC inhibition has been recognized as an attractive therapeutic strategy for AD [4, 5, 7].”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [4] Falkenberg KJ, Johnstone RW. Histone deacetylases and their inhibitors in cancer, neurological diseases and immune disorders. Nat Rev Drug Discov. 2014;13(9):673–91. source
    • cites [5] Burns AM, Graff J. Cognitive epigenetic priming: leveraging histone acetylation for memory amelioration. Curr Opin Neurobiol. 2021;67:75–84. source
    • cites [7] Kumar V, Kundu S, Singh A, Singh S. Understanding the role of histone deacetylase and their inhibitors in neurodegenerative disorders: current targets and future perspective. Curr Neuropharmacol. 2022;20(1):158–78. source

  • May combine two claims.

    Premisecitation

    CI-994 is an orally bioavailable specific class I HDAC inhibitor which can cross the blood-brain barrier.

    “CI-994 is an orally bioavailable specific class I HDAC inhibitor which can across the blood–brain barrier [16, 17].”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [16] Riva L, Blaney SM, Dauser R, Nuchtern JG, Durfee J, McGuffey L, et al. Pharmacokinetics and cerebrospinal fluid penetration of CI-994 (N-acetyldinaline) in the nonhuman primate. Clin Cancer Res. 2000;6(3):994–7.
    • cites [17] Graff J, Joseph NF, Horn ME, Samiei A, Meng J, Seo J, et al. Epigenetic priming of memory updating during reconsolidation to attenuate remote fear memories. Cell. 2014;156(1–2):261–76. source

      The Results section (“By Using HDAC2-Targeting HDAC Inhibitors, Remote Fear Memories Can Be Attenuated”; page unavailable) identifies CI-994 as a selective class I HDAC inhibitor and reports substantial brain levels after systemic administration, supporting its ability to reach the brain. It does not establish oral bioavailability: the reported administration was intraperitoneal. (LLM, conf=0.90)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Pan-HDAC inhibitors, which are effective in AD models, have been associated with unfavorable side effects and barely translated into clinical trials.

    “Pan-HDAC inhibitors, which are effective in AD models, have been associated with unfavorable side effects and barely translated into clinical trials [62].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [62] Yang SS, Zhang R, Wang G, Zhang YF. The development prospection of HDAC inhibitors as a potential therapeutic direction in Alzheimer’s disease. Transl Neurodegener. 2017;6:19. source

  • May combine two claims.

    Premisecitation

    β-catenin interacts with HDAC1 and attenuates its enzymatic activity.

    “Finally and importantly, β-catenin interacts with HDAC1 and attenuates its enzymatic activity [47].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [47] Billin AN, Thirlwell H, Ayer DE. Beta-catenin-histone deacetylase interactions regulate the transition of LEF1 from a transcriptional repressor to an activator. Mol Cell Biol. 2000;20(18):6882–90. source

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

    Premisecitation

    NRTN displays neuroprotection through enhancing dopaminergic neuronal survival and behavioural function in animal models.

    “NRTN displays neuroprotection through enhancing dopaminergic neuronal survival and behavioural function in animal models [73, 74], and adeno-associated virus serotype 2 (AAV-2) vector encoding the human NRTN gene (CERE120) has been developed to potentially provide clinical benefits in Parkinson’s disease [75].”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [73] Horger BA, Nishimura MC, Armanini MP, Wang LC, Poulsen KT, Rosenblad C, et al. Neurturin exerts potent actions on survival and function of midbrain dopaminergic neurons. J Neurosci. 1998;18(13):4929–37. source
    • cites [74] Fjord-Larsen L, Johansen JL, Kusk P, Tornoe J, Gronborg M, Rosenblad C, et al. Efficient in vivo protection of nigral dopaminergic neurons by lentiviral gene transfer of a modified Neurturin construct. Exp Neurol. 2005;195(1):49–60. source

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

    Premisecitation

    Drug repurposing can accelerate the identification of new therapeutic agents for AD.

    “Drug repurposing can accelerate the identification of new therapeutic agents for AD [80].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [80] Ballard C, Aarsland D, Cummings J, O’Brien J, Mills R, Molinuevo JL, et al. Drug repositioning and repurposing for Alzheimer disease. Nat Rev Neurol. 2020;16(12):661–73. source

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

    Premisecitation

    CACNA1G loss-of-function mutations are associated with schizophrenia and intellectual disability.

    “The thalamus-enriched voltage-dependent calcium channel CaV3.1 encoded by the Wnt target CACNA1G is the key mediator of thalamocortical connectivity, and putative loss-of-function mutations of CACNA1G have been associated with schizophrenia and intellectual disability [65–67].”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [65] Alazami AM, Patel N, Shamseldin HE, Anazi S, Al-Dosari MS, Alzahrani F, et al. Accelerating novel candidate gene discovery in neurogenetic disorders via whole-exome sequencing of prescreened multiplex consanguineous families. Cell Rep. 2015;10(2):148–61. source
    • cites [66] Singh T, Poterba T, Curtis D, Akil H, Al Eissa M, Barchas JD, et al. Rare coding variants in ten genes confer substantial risk for schizophrenia. Nature. 2022;604(7906):509–16. source

      The Discussion reports that protein-truncating variants (putative loss-of-function variants) in CACNA1G increase schizophrenia risk. It attributes the developmental-disorder and intellectual-disability risk instead to de novo missense mutations, and says the two variant types may have different functional effects. Thus the source does not support attributing both associations to loss-of-function mutations. (LLM, conf=0.90)

      Experimental — accuracy not yet measured
    • cites [67] Kim JI, Miura Y, Li MY, Revah O, Selvaraj S, Birey F, Meng X, Thete MV, Pavlov SD, Andersen J, et al. Human assembloids reveal the consequences of CACNA1G gene variants in the thalamocortical pathway. Neuron. 2024;112(24):4048-4059.e7. source

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

    Premisecitation

    Endogenous Wnt inhibitory protein NLK negatively regulates Wnt/β-catenin signaling via mediating HDAC1 phosphorylation.

    “Fourthly, endogenous Wnt inhibitory protein NLK negatively regulates Wnt/β-catenin signaling via mediating HDAC1 phosphorylation [44].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [44] Masoumi KC, Daams R, Sime W, Siino V, Ke H, Levander F, et al. NLK-mediated phosphorylation of HDAC1 negatively regulates Wnt signaling. Mol Biol Cell. 2017;28(2):346–55. source

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

    Premisecitation

    Wnt target NEUROD1 is important for adult neurogenesis and the survival and maturation of adult-born neurons.

    “Wnt target NEUROD1 is important for adult neurogenesis and the survival and maturation of adult-born neurons [54, 64].”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [54] Kuwabara T, Hsieh J, Muotri A, Yeo G, Warashina M, Lie DC, et al. Wnt-mediated activation of NeuroD1 and retro-elements during adult neurogenesis. Nat Neurosci. 2009;12(9):1097–105. source
    • cites [64] Gao Z, Ure K, Ables JL, Lagace DC, Nave KA, Goebbels S, et al. Neurod1 is essential for the survival and maturation of adult-born neurons. Nat Neurosci. 2009;12(9):1090–2. source

      The paper’s Abstract reports that Neurod1 is required for the survival and maturation of adult-born neurons and identifies it as a regulator of adult neurogenesis. Our copy does not identify Neurod1 as a Wnt target (searched: “Wnt target Neurod1” and “Wnt”), so that part of the sentence is not supported by this source. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

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

    Premisecitation

    AAV-2 vector encoding human NRTN gene (CERE120) has been developed to potentially provide clinical benefits in Parkinson's disease.

    “NRTN displays neuroprotection through enhancing dopaminergic neuronal survival and behavioural function in animal models [73, 74], and adeno-associated virus serotype 2 (AAV-2) vector encoding the human NRTN gene (CERE120) has been developed to potentially provide clinical benefits in Parkinson’s disease [75].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [75] Chu Y, Bartus RT, Manfredsson FP, Olanow CW, Kordower JH. Long-term post-mortem studies following neurturin gene therapy in patients with advanced Parkinson’s disease. Brain. 2020;143(3):960–75. source

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

    Premisecitation

    SLC7A10 is a potential rare gene associated with human startle disease and developmental delay.

    “Although the function of neutral amino acid transporter SLC7A10 in CNS remains to be elucidated, studies with SLC7A10 knock-out mice and dysfunctional SLC7A10 mutations reveal that SLC7A10 is a potential rare gene associated with human startle disease and developmental delay [76, 77].”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [76] Safory H, Neame S, Shulman Y, Zubedat S, Radzishevsky I, Rosenberg D, et al. The alanine-serine-cysteine-1 (Asc-1) transporter controls glycine levels in the brain and is required for glycinergic inhibitory transmission. EMBO Rep. 2015;16(5):590–8. source

      The paper’s summary states that Asc-1 (SLC7A10) is a candidate for hyperekplexia, or human startle disease, based on its knockout-mouse findings (Safory et al., summary; page unavailable). It does not report dysfunctional SLC7A10 mutations in humans or link the gene to developmental delay. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [77] Drehmann P, Milanos S, Schaefer N, Kasaragod VB, Herterich S, Holzbach-Eberle U, Harvey RJ, Villmann C. Dual role of dysfunctional Asc-1 transporter in distinct human pathologies, human startle disease, and developmental delay. eNeuro. 2023;10(11):ENEURO.0263-23.2023. source

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

    Premisecitation

    LRP6 turnover is regulated by the R-spondin/Lgr5/Rnf43 module.

    “The LRP6 turnover is regulated by the Dickkopf1/Kremen pathway [81] and R-spondin/Lgr5/Rnf43 module [82].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [82] de Lau W, Peng WC, Gros P, Clevers H. The R-spondin/Lgr5/Rnf43 module: regulator of Wnt signal strength. Genes Dev. 2014;28(4):305–16. source

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

    Findingown data

    CI-994 increases histone H3 acetylation and β-catenin in neuronal SH-SY5Y cells after 24 h treatment.

    “Western blotting analyses of histone H3 acetylation and β-catenin in neuronal SH-SY5Y cells after 24 h treatment. The cells were treated with CI-994 at the indicated concentrations for 24 h, the levels of acetyl histone H3K9 (D, E), total histone H3 (F, G), and β-catenin (H, I) were examined by Western blotting and quantified.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CI-994 greatly increased Wnt reporter activity in Wnt-3A-expressing HEK293 cells with an EC50 value around 5.8 μM.

    “We demonstrated that CI-994 greatly increased Wnt reporter activity in Wnt-3A-expressing HEK293 cells with an EC50 value around 5.8 µM (Fig. 1B)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    LGK974 abolished CI-994-induced elevation of β-catenin level in iPSC-derived cerebral organoids, although there was no statistical significance due to sample variance effects.

    “LGK974 abolished CI-994-induced elevation of β-catenin level (Fig. 5B and D) in the iPSC-derived cerebral organoids, although there was no statistical significance due to the sample variance effects.”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    CI-994 dramatically reduced tau phosphorylation in iPSC-derived neurons carrying APOE ε4/ε4.

    “CI-994 significantly enhanced the levels of total β-catenin, and dramatically reduced tau phosphorylation in the iPSC-derived neurons (Fig. 6A, E, F).”
    Quote in the paperSubstantial claim

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

    Study design and conductown data

    The heterozygous MAPT p.P301L mutation in iPSC line MC0071 was verified by TaqMan genotyping assay.

    “the heterozygous MAPT p.P301L mutation in iPSC line MC0071 was verified by the TaqMan genotyping assay with probe rs63751273 from Thermo Fisher Scientific.”
    Quote in the paperNot a substantial claim

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

    Interpretationown data

    CI-994 is a dual modulator of class I HDACs and Wnt/β-catenin signaling for the treatment of Alzheimer's disease.

    “CI-994 is a dual modulator of class I HDACs and Wnt/β-catenin signaling.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The effects of CI-994 observed in cell lines and patient-specific iPSC-derived cerebral organoids predict therapeutic effects in human Alzheimer's disease.

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

    Study design and conductown data

    The APOE genotype of iPSC lines MC0020 and MC0071 was verified by TaqMan genotyping assay.

    “The APOE genotype of iPSC lines MC0020 and MC0071 was verified by the TaqMan genotyping assay with probes rs7412 and rs429358 from Thermo Fisher Scientific”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    CI-994 is a dual modulator for class I HDAC inhibition and Wnt/β-catenin signaling activation.

    “Together, these findings indicate that CI-994 is a dual modulator for class I HDAC inhibition and Wnt/β-catenin signaling activation.”
    • Also stated in: § CI-994 is not only a class I HDAC inhibitor but also a Wnt/β-catenin signaling activator · plain
      “CI-994 is not only a class I HDAC inhibitor but also a Wnt/β-catenin signaling activator”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Study design and conductown data

    The pluripotency of iPSC lines was confirmed by three germ layer differentiation and immunostaining of pluripotency markers.

    “To confirm the pluripotency of iPSCs, three germs layer differentiation was performed with the STEMdiff Trilineage Differentiation kit (Stemcell Technologies) according to manufacturer’s instructions.”
    • Also stated in: § Generation of iPSCs from human skin fibroblasts and trilineage differentiation of human iPSCs · plain
      “The pluripotency and differentiation capacity of iPSC lines was examined by immunostaining of pluripotency markers T-antigen-related antigen 1–60 (TRA-1–60) and Nanog, and was further validated by immunostaining of three germ layer-specific markers SOX17 for Endoderm, Brachyury for Mesoderm, and Nestin for Ectoderm”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    BML-284 is an agonist of Wnt/β-catenin signaling with an EC50 of 0.7 μM of Wnt reporter activity in HEK293 cells.

    “BML-284 is an agonist of Wnt/β-catenin signaling with an EC50 of 0.7 µM of Wnt reporter activity in HEK293 cells [41].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [41] Liu J, Wu X, Mitchell B, Kintner C, Ding S, Schultz PG. A small-molecule agonist of the Wnt signaling pathway. Angew Chem Int Ed Engl. 2005;44(13):1987–90. source

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

    Premisecitation

    CI-994 is an analog of niclosamide.

    “Interesting, a recent study has shown that CI-994 (Fig. 1A), an analog of niclosamide and an inhibitor of class I HDACs [39], is able to enhance the TCF/LEF reporter activity of Wnt/β-catenin signaling [40], suggesting that CI-994 could display an opposite effect on Wnt/β-catenin signaling.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [39] Moradei OM, Mallais TC, Frechette S, Paquin I, Tessier PE, Leit SM, et al. Novel aminophenyl benzamide-type histone deacetylase inhibitors with enhanced potency and selectivity. J Med Chem. 2007;50(23):5543–6. source

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

    Study design and conductown data

    Karyotyping of the iPSC lines was performed.

    “Karyotyping of the iPSC lines was performed by Mayo Clinic Cytogenetics Core.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    Karyotyping test confirmed that number and appearance of chromosomes were normal in iPSC line MC0071.

    “Karyotyping test confirmed that the number and appearance of chromosomes were normal in iPSC line MC0071 (Fig. S4).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    CI-994 enhances histone acetylation and activates Wnt/β-catenin signaling in patient-specific iPSC-derived cerebral organoids.

    “Together, these findings indicate that CI-994 enhances histone acetylation and activates Wnt/β-catenin signaling in patient-specific iPSC-derived cerebral organoids.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    OMG is one of the three major myelin inhibitors that signal through neuronal receptors to reorganize the cytoskeleton and inhibit axon growth.

    “OMG is one of the three major myelin inhibitors that signal through neuronal receptors to reorganize the cytoskeleton and inhibit axon growth [61].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [61] Zheng B, Tuszynski MH. Regulation of axonal regeneration after mammalian spinal cord injury. Nat Rev Mol Cell Biol. 2023;24(6):396–413. source

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

    Premisecitation

    NRTN is a member of the glial cell-line derived neurotrophic factor family which promotes neuronal survival.

    “NRTN is a member of the glial cell-line derived neurotrophic factor family which promotes neuronal survival [59].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [59] Kotzbauer PT, Lampe PA, Heuckeroth RO, Golden JP, Creedon DJ, Johnson EM Jr, et al. Neurturin, a relative of glial-cell-line-derived neurotrophic factor. Nature. 1996;384(6608):467–70. source

  • May combine two claims.

    Interpretationreasoning

    CI-994 could upregulate expression of NEUROD1, CACNA1G, NRGN, NRTN and SLC7A10 and downregulate expression of OMG to exert its neuroprotective actions.

    “Therefore, CI-994 could upregulate the expression of NEUROD1, CACNA1G, NRGN, NRTN and SLC7A10 and downregulate the expression of OMG to exert its neuroprotective actions.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    CI-994 is a brain-penetrant inhibitor of class I HDACs.

    “In conclusion, CI-994, a brain penetrant and orally bioavailable inhibitor of class I HDACs, is also an activator of Wnt/β-catenin signaling.”
    • Also stated in: § Limitations of the study · plain
      “CI-994 is a brain penetrant.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [16] Riva L, Blaney SM, Dauser R, Nuchtern JG, Durfee J, McGuffey L, et al. Pharmacokinetics and cerebrospinal fluid penetration of CI-994 (N-acetyldinaline) in the nonhuman primate. Clin Cancer Res. 2000;6(3):994–7.

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

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

    Interpretationreasoning

    There is a potential therapeutic-positive feedback loop between class I HDAC inhibition and Wnt/β-catenin signaling activation.

    “Therefore, the in vivo therapeutic effects of CI-994 could be mainly attributed to class I HDAC inhibition, but it should be noticed that there is a potential therapeutic-positive feedback loop between class I HDAC inhibition and Wnt/β-catenin signaling activation, and that CI-994 at a concentration lower than the EC50 value is still able to activate Wnt/β-catenin signaling.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationnone given

    CI-994 is a potent dual modulator for class I HDACs and Wnt/β-catenin signaling.

    “Second, although CI-994 is a potent dual modulator for class I HDACs and Wnt/β-catenin signaling, it is unclear whether HDAC inhibition and Wnt/β-catenin signaling activation contribute equally to the in vivo therapeutic effects of CI-994.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    LRP6 turnover is regulated by the Dickkopf1/Kremen pathway.

    “The LRP6 turnover is regulated by the Dickkopf1/Kremen pathway [81] and R-spondin/Lgr5/Rnf43 module [82].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [81] Mao B, Wu W, Davidson G, Marhold J, Li M, Mechler BM, et al. Kremen proteins are Dickkopf receptors that regulate Wnt/beta-catenin signalling. Nature. 2002;417(6889):664–7. source

  • May combine two claims.

    Premisecitation

    Peak plasma and CSF CI-994 concentrations after an 80-mg/m2 intravenous dose were 23.3 μM with t1/2 7.4 h and 3.4 μM with t1/2 12.9 h, respectively, in nonhuman primates.

    “The peak plasma and cerebrospinal fluid (CSF) CI-994 concentrations after an 80-mg/m2 intravenous dose were 23.3 µM with a terminal half-life (t1/2) at 7.4 h and 3.4 µM with a t1/2 at 12.9 h, respectively, in nonhuman primates [16].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [16] Riva L, Blaney SM, Dauser R, Nuchtern JG, Durfee J, McGuffey L, et al. Pharmacokinetics and cerebrospinal fluid penetration of CI-994 (N-acetyldinaline) in the nonhuman primate. Clin Cancer Res. 2000;6(3):994–7.

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

46 of 46

  • The paper relies on this without support.

    The iPSC-derived neurons and cerebral organoids carrying APOE ε4/ε4 or MAPT p.P301L mutation recapitulate key aspects of Alzheimer's disease pathology and are predictive of therapeutic effects in human AD.

    “The therapeutic effects of CI-994 were examined in patient-specific iPSC-derived neurons and cerebral organoids carrying APOE ε4/ε4 genotype or MAPT p.P301L mutation.”
    “Human skin biopsies from a patient with AD (MC0020; female, 71 years old) carrying APOE ε4/ε4 genotype and a patient with Parkinsonism linked to chromosome 17 (FTDP-17) at risk (MC0071; male, 47 years old) carrying APOE ε3/ε3 genotype and heterozygous MAPT p.P301L mutation were obtained from Mayo Clinic patients under IRB protocol with patient consent for research, which was approved by the Mayo”

  • The paper relies on this without support.

    The effects of CI-994 observed in cell lines and patient-specific iPSC-derived cerebral organoids predict therapeutic effects in human Alzheimer's disease.

    “To explore the therapeutic effects of CI-994 in AD”

  • The paper relies on this without support.

    The iPSC-derived neurons and cerebral organoids carrying APOE ε4/ε4 or MAPT p.P301L mutation recapitulate AD-relevant tau pathology and can predict therapeutic effects in AD.

    “CI-994 significantly inhibits tau phosphorylation in patient-specific iPSC-derived neurons and cerebral organoids carrying APOE ε4/ε4 or MAPT p.P301L mutation.”

  • The paper relies on this without support.

    The molecular and cellular effects of CI-994 observed in these models translate to clinical benefit in AD patients.

    “Therefore, CI-994 is a potential therapeutic candidate for alleviating both amyloid and tau pathologies in AD.”

  • The paper relies on this without support.

    CI-994 is orally bioavailable.

    “In conclusion, CI-994, a brain penetrant and orally bioavailable inhibitor of class I HDACs, is also an activator of Wnt/β-catenin signaling.”

  • The paper relies on this without support.

    In vitro IC50/EC50 potency differences predict the relative in vivo contribution of HDAC inhibition and Wnt/β-catenin signaling activation.

    “Notably, the IC50 values of HDAC 1, 2 and 3 inhibition are greatly lower than the EC50 value of Wnt/β-catenin signaling activation.”
    “Therefore, the in vivo therapeutic effects of CI-994 could be mainly attributed to class I HDAC inhibition, but it should be noticed that there is a potential therapeutic-positive feedback loop between class I HDAC inhibition and Wnt/β-catenin signaling activation, and that CI-994 at a concentration lower than the EC50 value is still able to activate Wnt/β-catenin signaling.”
6 of 6

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.

  • Niclosamide HDAC11 mean does not match its duplicate values

    Minor

    The mean of the two niclosamide HDAC11 activity values is 119.43%, not the reported 103.43%.

    Reported 103.43 · recomputed 119.43

  • LGK974 is labeled both a Wnt activator and a Wnt inhibitor

    Minor

    The materials section calls LGK974 a Wnt activator, whereas the results identify it as a specific Wnt inhibitor.

  • Figure 7 title identifies organoids, but its caption describes neurons

    Minor

    Figure 7 is titled as an experiment in cerebral organoids, while its caption says neurons were collected after the final MEA measurement.

  • HDAC10 appears in the table but not the stated profiling panel

    Minor

    The methods specify a ten-HDAC panel comprising HDAC1-9 and HDAC11, but Table 1 also reports results for HDAC10.

4 of 4

Cites

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Cited by

Sorted by citations

Authors
Lu, W., Kawatani, K., Ren, Y., Nambara, T., Jia, L., Jeevaratnam, S., Lee, E., Martinez, P. R., Izhar, T., Wang, N., Raulin, A.-C., Wszolek, Z. K., Bu, G., Kanekiyo, T., & Li, Y.
Affiliations
  • Mayo Clinic (US)
  • Mayo Clinic in Florida (US)
Corresponding author
Li, Yonghe
ORCID
Venue
Alzheimer's Research & Therapy
Publisher
Springer Science and Business Media LLC
Published
2026-02-07
Volume
18
Issue
1
ISSN
1758-9193 (ISSN-L)
DOI
10.1186/s13195-026-01982-0
PMC
PMC12977471
OpenAlex
W7128295769
Semantic Scholar
34f82b4488c495572890497bdd221624ee713de0
Cited by
4
References
80
Citations by year
2026: 4
FWCI
15.71
Type
journal article
Language
en
Open access
Licence
Locations
  • Alzheimer s Research & Therapy (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, cc-by-nc-nd)
  • Rare & Special e-Zone (The Hong Kong University of Science and Technology) (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
NIH, Florida Department of Health Ed and Ethel Moore Alzheimer’s Disease Research Program (R21AG065653, R01AG078615, 24A07)
Topics
  • Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Histone Deacetylase Inhibitors Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Wnt/β-catenin signaling in development and cancer
Keywords
CI-994, class I histone deacetylase inhibitors, Wnt/β-catenin signaling, Alzheimer's disease, LRP6, histone acetylation, tau phosphorylation, APOE ε4, synaptic dysfunction, LGK974, epigenetic therapy, blood-brain barrier
MeSH terms
Aged, Alzheimer Disease, Benzamides, Brain, Female, Histone Deacetylases, Humans, Male, Middle Aged, Neurons, Organoids, Phenylenediamines, Phosphorylation, tau Proteins, Cell Line, Tumor, beta Catenin, Histone Deacetylase Inhibitors, Induced Pluripotent Stem Cells, HEK293 Cells, Wnt Signaling Pathway

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 2026-10-18.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-18.

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.

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