Inhibition of ACAT as a Therapeutic Target for Alzheimer's Disease Is Independent of ApoE4 Lipidation

Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

ApoE is the only lipoprotein-competent apolipoprotein expressed in the brain, where it is secreted primarily by astrocytes.

“Apolipoprotein E (apoE), the protein encoded by the APOE gene, is the only lipoprotein-competent apolipoprotein expressed in the brain, where it is secreted primarily by astrocytes [11, 12].”

• cites [11] Elliott, 2010, Apolipoproteins in the brain: implications for neurological and psychiatric disorders, Clin Lipidol sourceDisputed

  The ApoE section supports astrocytes as the primary source of brain ApoE. But the “Unique features of CNS lipid/cholesterol & apolipoprotein metabolism” section describes ApoJ-containing lipoprotein particles and identifies astrocytes as a major source of both ApoJ and ApoE in the CSF; the ApoJ/clusterin section also says ApoJ is produced in the brain. Thus the paper contradicts the claim that ApoE is the only lipoprotein-competent apolipoprotein expressed there. (LLM, conf=0.88)

  Checked by a reasoning model — may be wrong.

  Experimental — accuracy not yet measured
• cites [12] LaDu, 1998, Nascent astrocyte particles differ from lipoproteins in CSF, J Neurochem sourceNot checked — no full text of the source

Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

RXR agonists induce an increase in ABCA1 expression with an increase in apoE4 lipidation and a decrease in both total Aβ42 and soluble Aβ in EFAD-Tg mice.

“We previously demonstrated that RXR agonists induce an increase in ABCA1 expression with an increase in apoE4 lipidation and a decrease in both total Aβ42 and soluble Aβ in EFAD (5xFAD+/-/APOE4+/+)-Tg mice [13].”

• cites [13] Youmans, 2012, APOE4-specific changes in abeta accumulation in a new transgenic mouse model of Alzheimer disease, J Biol Chem sourceDisputed

  The Discussion (p. 41783) explicitly says that drug intervention trials had not yet been performed in EFAD mice. It describes an RXR-agonist result in other mice expressing mouse apoE, not an increase in ABCA1 or apoE4 lipidation and a decrease in Aβ in treated EFAD mice. (LLM, conf=0.94)

  Checked by a reasoning model — may be wrong.

  Experimental — accuracy not yet measured

It may state two separate claims as one.

The lipid droplets are rich in CE and poor in TAG, thus not supporting the formation of apoE4-TAG particles.

“evidence suggests that the LD are rich in CE and poor in TAG, thus not supporting the formation of apoE4-TAG particles [130].”

• cites [130] Lindner, 2022, Isoform- and cell-state-specific lipidation of ApoE in astrocytes, Cell Rep sourceDisputed

  The section “Fatty astrocytes secrete ApoE-containing, triacylglycerol-rich particles” says TAG mainly accumulates in lipid droplets and reports that fatty astrocytes secrete TAG-loaded APOE4 particles. The Discussion (p. 9) likewise proposes that ApoE can be loaded with TAG where lipid droplets form. The paper therefore makes the opposite point; it does not describe the droplets as CE-rich and TAG-poor. (LLM, conf=0.88)

  Checked by a reasoning model — may be wrong.

  Experimental — accuracy not yet measured

Probably not a substantial claim.

Lipid dysregulation is involved in the pathogenesis and progression of neurodegenerative diseases including Alzheimer's disease.

“Lipid dysregulation is involved in the pathogenesis and progression of neurodegenerative diseases including Alzheimer’s disease (AD) [1–3].”

• cites [1] Ralhan, 2021, Lipid droplets in the nervous system, J Cell Biol sourceSupported
• cites [2] Picard, 2018, Alterations in cholesterol metabolism-related genes in sporadic Alzheimer's disease, Neurobiol Aging sourceNot checked — no full text of the source
• cites [3] Yin, 2022, Lipid metabolism and Alzheimer's disease: clinical evidence, mechanistic link and therapeutic promise, FEBS J sourcePartly supported

  The Introduction directly connects disrupted lipid metabolism with the pathogenesis and progression of Alzheimer’s disease. The lipid-homeostasis section also notes lipid droplets in other neurodegenerative brains, but it does not establish the same pathogenesis-and-progression claim for neurodegenerative diseases generally. Page numbers were not available in our copy. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Probably not a substantial claim. The paper’s conclusions may not depend on it.

Aβ peptides are produced primarily by neurons via sequential cleavage of APP by β- and γ-secretases at lipid rafts in the plasma membrane.

“Aβ peptides are produced primarily by neurons via the sequential cleavage of amyloid precursor protein (APP) by β- and γ-secretases (amyloidogenic pathway) at lipid rafts in the plasma membrane (PM) [35–37].”

• cites [35] Hampel, 2021, The beta-secretase BACE1 in Alzheimer's disease, Biol Psychiatry sourcePartly supported

  The Amyloidogenesis section says neurons produce the highest levels of Aβ and that BACE1 (β-secretase), together with γ-secretase, generates Aβ from APP. The paper also discusses BACE1 activity in lipid rafts, but it does not place the sequential cleavage specifically at plasma-membrane lipid rafts; the same section describes BACE1 processing of APP in endosomes. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [36] Cordy, 2003, Exclusively targeting beta-secretase to lipid rafts by GPI-anchor addition up-regulates beta-site processing of the amyloid precursor protein, Proc Natl Acad Sci U S A sourceNot checked — no full text of the source
• cites [37] Wahrle, 2002, Cholesterol-dependent gamma-secretase activity in buoyant cholesterol-rich membrane microdomains, Neurobiol Dis sourceNot checked — no full text of the source

Probably not a substantial claim. The paper’s conclusions may not depend on it.

ApoE4 levels in brain are lower than apoE3 levels in both humans and transgenic mouse models.

“Although expressed at comparable levels, apoE4 levels in brain are lower than apoE3 levels in both humans and Tg mouse models [13–15], suggesting that apoE4 is unstable and undergoes accelerated degradation, likely due to the observed reduced lipidation state of apoE4-lipoproteins compared to apoE3-lipoproteins [16–23].”

• cites [13] Youmans, 2012, APOE4-specific changes in abeta accumulation in a new transgenic mouse model of Alzheimer disease, J Biol Chem sourceSupported
• cites [14] Beffert, 1999, Apolipoprotein E and beta-amyloid levels in the hippocampus and frontal cortex of Alzheimer’s disease subjects are disease-related and apolipoprotein E genotype dependent, Brain Res sourceNot checked — no full text of the source
• cites [15] Sullivan, 2011, Reduced levels of human apoE4 protein in an animal model of cognitive impairment, Neurobiol Aging sourceNot checked — no full text of the source

It may state two separate claims as one.

Mouse apoE and human APP combine in FAD-Tg mice to produce severe Aβ pathology, which is reduced by knocking out mouse APOE, further reduced by knocking in human APOE4, and again reduced with human APOE3 knock-in.

“Importantly, m-apoE and h-APP combine in FAD-Tg mice to produce severe Aβ pathology, pathology that is reduced by knocking-out (KO) the m-APOE, further reduced by knocking-in (KI) h-APOE4, and again reduced with h-APOE3-KI (review [79–87]).”

• cites [79] Tai, 2017, EFAD transgenic mice as a human APOE relevant preclinical model of Alzheimer's disease, J Lipid Res sourcePartly supported

  Table 1 (‘Amyloid plaque deposition is not the same as soluble Aβ peptide’; page unavailable) shows less Aβ pathology in FAD-Tg mice lacking APOE than in FAD-Tg mice with mouse APOE. The EFAD results (‘APOE modulated AD symptoms exhibited in EFAD mice’; page unavailable) report the ordering 5xFAD > E4FAD > E3FAD. But the paper does not establish the claimed successive reduction from APOE knockout to human APOE4 knock-in in the same model: Table 1’s knockout comparison is with a human-APOE transgenic model, while its APOE targeted-replacement comparison uses 5xFAD mice. (LLM, conf=0.85)

  Experimental — accuracy not yet measured
• cites [80] Trommer, 2004, ApoE isoform affects LTP in human targeted replacement mice, NeuroReport sourceNot checked — no full text of the source
• cites [81] Trommer, 2005, ApoE isoform-specific effects on LTP: blockade by oligomeric amyloid-beta1-42, Neurobiol Dis sourceNot checked — no full text of the source
• cites [82] Bien-Ly, 2011, C-terminal-truncated apolipoprotein (apo) E4 inefficiently clears amyloid-beta (abeta) and acts in concert with abeta to elicit neuronal and behavioral deficits in mice, Proc Natl Acad Sci U S A sourceNot checked — no full text of the source
• cites [83] Zhu, 2012, APOE genotype alters glial activation and loss of synaptic markers in mice, Glia sourcePartly supported

  The Introduction reports that APOE4 increases amyloid levels in animal models and discusses apoE’s role in Aβ deposition. The Methods (Animals) describe APOE knock-out and human APOE knock-in mice, but the paper does not report FAD-Tg mice expressing human APP or the claimed stepwise reductions in Aβ pathology. (LLM, conf=0.78)

  Experimental — accuracy not yet measured
• cites [84] Liao, 2015, Murine versus human apolipoprotein E4: differential facilitation of and co-localization in cerebral amyloid angiopathy and amyloid plaques in APP transgenic mouse models, Acta Neuropathol Commun sourcePartly supported

  The Introduction says APP-transgenic mice lacking apoE have markedly reduced fibrillar Aβ deposition. The Results report fewer plaques when mouse apoE is replaced with human apoE4, but more cerebral amyloid angiopathy. Neither section establishes the claimed sequence of further reductions from apoE knockout to apoE4 knock-in to apoE3 knock-in; the study does not test an apoE3 knock-in group. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [85] Fagan, 2002, Human and murine ApoE markedly alters A beta metabolism before and after plaque formation in a mouse model of Alzheimer's disease, Neurobiol Dis sourceNot checked — no full text of the source
• cites [86] Fryer, 2005, Human apolipoprotein E4 alters the amyloid-beta 40:42 ratio and promotes the formation of cerebral amyloid angiopathy in an amyloid precursor protein transgenic model, J Neurosci sourcePartly supported

  The Introduction reports that APPsw mice with murine apoE develop plaques and CAA, while mice lacking murine apoE have predominantly nonfibrillar deposits and no CAA. The Results and Discussion report less amyloid deposition with human apoE4 than with murine apoE, and less with apoE3 than with apoE4. They do not support the claimed further reduction from apoE knockout to apoE4 knock-in: apoE4 mice developed substantial CAA, whereas knockout mice did not. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [87] Oddo, 2009, Genetically altering abeta distribution from the brain to the vasculature ameliorates tau pathology, Brain Pathol sourcePartly supported

  The Results (section ‘Expression of apoE4 leads to a reduction in parenchyma Aβ deposits and Aβ levels’; page unavailable) report reduced parenchymal Aβ after replacing mouse apoE with human apoE4 in 3xTg-AD mice. The paper does not test an APOE knockout alone or an APOE3 knock-in as steps in the claimed sequence. It also reports increased vascular Aβ with apoE4, so the reduction is specific to parenchymal pathology. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Probably not a substantial claim.

The APP processing pathway is highly regulated by mobilization of cholesterol between lipid rafts and the plasma membrane.

“The APP processing pathway is highly regulated by the mobilization of cholesterol between the lipid rafts and PM [41, 42].”

• cites [41] Bhattacharyya, 2021, Axonal generation of amyloid-beta from palmitoylated APP in mitochondria-associated endoplasmic reticulum membranes, Cell Rep sourcePartly supported

  The Discussion (page not provided) reports that changes in MAM assembly alter palmitoylated APP trafficking, β-secretase cleavage, and Aβ generation, supporting regulation of APP processing. It does not show that mobilization of cholesterol between lipid rafts and the plasma membrane is the regulating mechanism. (LLM, conf=0.83)

  Experimental — accuracy not yet measured
• cites [42] Chauhan, 2003, Membrane dynamics, cholesterol homeostasis, and Alzheimer's disease, J Lipid Res sourceNot checked — no full text of the source

It may state two separate claims as one.

Inhibiting ACAT increases the free cholesterol pool and facilitates lipid secretion to extracellular apoE-containing lipoproteins.

“Thus, inhibiting ACAT increases the FC pool and facilitates lipid secretion to extracellular apoE-containing lipoproteins.”

• Also stated in: § Introduction · plain

  “By repurposing a cardiovascular drug that inhibits ACAT, specifically avasimibe (AVAS), LD formation is inhibited, allowing FC build-up, thus promoting ABCA1-mediated cholesterol efflux to nascent apoE4-lipoproteins [57–63].”

• cites [57] Sugimoto, 2004, An inhibitor of acylCoA: cholesterol acyltransferase increases expression of ATP-binding cassette transporter A1 and thereby enhances the ApoA-I-mediated release of cholesterol from macrophages, Biochim Biophys Acta sourceNot checked — no full text of the source
• cites [58] Rodriguez, 2002, Anti-atherogenic effects of the acyl-CoA:cholesterol acyltransferase inhibitor, avasimibe (CI-1011), in cultured primary human macrophages, Atherosclerosis sourceNot checked — no full text of the source
• cites [59] Kellner-Weibel, 2003, Cytotoxic cellular cholesterol is selectively removed by apoA-I via ABCA1, Atherosclerosis sourceNot checked — no full text of the source
• cites [60] Huang, 2001, Sterol efflux mediated by endogenous macrophage ApoE expression is independent of ABCA1, Arterioscler Thromb Vasc Biol sourceNot checked — no full text of the source
• cites [61] An, 2008, Inhibition of acyl-coenzyme A:cholesterol acyltransferase stimulates cholesterol efflux from macrophages and stimulates farnesoid X receptor in hepatocytes, Exp Mol Med sourcePartly supported

  In 'Decrement of CE mass dominates the negative effect of the accumulation of FC' (page unavailable), ACAT inhibition increased cellular free cholesterol and its secretion into the medium in acLDL-loaded THP-1 macrophages. The paper does not show that the secreted lipid entered extracellular apoE-containing lipoproteins; 'ACAT inhibition triggers changes in expression of genes involved in cholesterol catabolism and mobilization' instead reports that Apoe mRNA was unaffected. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [62] Yamauchi, 2004, Intracellular cholesterol mobilization involved in the ABCA1/apolipoprotein-mediated assembly of high density lipoprotein in fibroblasts, J Lipid Res sourcePartly supported

  The section ‘Effect of ACAT enzyme activity on apoA-I-mediated cholesterol release’ reports that ACAT inhibition increased cholesterol and phospholipid release mediated by apoA-I, not apoE. Table 1 in ‘Measurement of the free cholesterol pool available for ACAT’ does not establish the claimed increase in free cholesterol: the ACAT-deficient cells had less measured free cholesterol than their parental cells. The paper therefore supports increased lipid release under its tested conditions, but not the specified apoE-containing recipients or the asserted increase in the free-cholesterol pool. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [63] Schmitz, 1985, Regulation of high density lipoprotein receptors in cultured macrophages: role of acyl-CoA:cholesterol acyltransferase, EMBO J sourceNot checked — no full text of the source

It may state two separate claims as one.

ACAT activity and lipid droplet levels increase with age, AD, and APOE4, suggesting low free cholesterol levels that will limit apoE lipidation and its downstream effects.

“ACAT activity and LD levels increase with age [64–66], AD, and APOE4 [67, 68], suggesting low levels of FC that will limit apoE lipidation and its downstream effects.”

• cites [64] Stahlberg, 1991, Age-related changes in the metabolism of cholesterol in rat liver microsomes, Lipids sourceNot checked — no full text of the source
• cites [65] Geltinger, 2020, The transfer of specific mitochondrial lipids and proteins to lipid droplets contributes to proteostasis upon stress and aging in the eukaryotic model system Saccharomyces cerevisiae, Geroscience sourcePartly supported

  The Conclusion reports increased lipid content in lipid droplets during aging in yeast, which supports an age-related change in droplet composition. It does not establish that lipid-droplet levels or ACAT activity increase with age. The AD and APOE4 portions are assigned to other citations, and this paper does not address the proposed free-cholesterol–apoE mechanism. (LLM, conf=0.83)

  Experimental — accuracy not yet measured
• cites [66] Beas, 2020, Independent regulation of age associated fat accumulation and longevity, Nat Commun sourcePartly supported

  The Results section, “Lipid droplets accumulate during yeast replicative aging” (page unavailable), reports a 7.2-fold increase in lipid droplets from young to old yeast cells. That supports the age-associated lipid-droplet portion of the citation, but the paper does not establish an age-related increase in ACAT activity or the proposed free-cholesterol and apoE-lipidation consequences. The AD and APOE4 portions are attached to other citations in the printed sentence. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [67] Sienski, 2021, APOE4 disrupts intracellular lipid homeostasis in human iPSC-derived glia, Sci Transl Med sourcePartly supported

  The Introduction reports greater lipid-droplet accumulation in APOE4 than APOE3 iPSC-derived astrocytes, and the Discussion notes intracellular lipid accumulation associated with AD. The Results section on cholesterol accumulation does not report increased ACAT activity or establish that free cholesterol is low and limits apoE lipidation; it instead reports greater cellular cholesterol in APOE4 astrocytes. The paper’s evidence is also from cell models, not an age-related comparison. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [68] Farmer, 2019, Apolipoprotein E4 alters astrocyte fatty acid metabolism and lipid droplet formation, Cells sourcePartly supported

  The Introduction and Discussion report increased lipid droplets in AD, and the Conclusions report more lipid droplets in E4 than E3 astrocytes. The paper does not establish increased ACAT activity in AD or APOE4, or the proposed link from low free cholesterol to limited apoE lipidation. (LLM, conf=0.85)

  Experimental — accuracy not yet measured

It may state two separate claims as one.

Brains of AD patients and familial AD-transgenic mice have elevated cholesterol levels.

“In addition, brains of AD patients [9] and familial AD-transgenic (FAD-Tg) mice [4, 5] have elevated cholesterol levels.”

• cites [9] Feringa, 2021, Cholesterol and Alzheimer's disease; from risk genes to pathological effects, Front Aging Neurosci sourcePartly supported

  In “Cholesterol, APP Processing and Aβ Generation in Neurons” (page unavailable), the review reports increased cholesterol esters (CE) in human postmortem AD brain tissue and AD mouse models; “Cholesterol and Tau” also reports CE accumulation in familial-AD mouse models. These findings support altered cholesterol storage, but not the broader claim that brain cholesterol levels overall are elevated. The review additionally reports lower cholesterol in lipid rafts from AD brains. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [4] Chan, 2012, Comparative lipidomic analysis of mouse and human brain with Alzheimer disease, J Biol Chem sourcePartly supported

  The Results report elevated cholesterol esters in the forebrains of PS1-APP mice, with a weaker upward trend in APP mice. They do not establish elevated cholesterol levels generally across FAD-transgenic mice; the Discussion says the ester increase was virtually absent in PS1 mice. The AD-patient portion of the sentence is attached to a different citation. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [5] Tajima, 2013, Lipidomic analysis of brain tissues and plasma in a mouse model expressing mutated human amyloid precursor protein/tau for Alzheimer's disease, Lipids Health Dis sourcePartly supported

  The citation to this paper is attached to the claim about familial AD-transgenic mice. The Results and Discussion report increased brain levels of a specific cholesterol ester, 22:6ChE, in APP/tau mice compared with controls. They do not establish that overall brain cholesterol levels were elevated; the AD-patient portion of the sentence is attributed to a different citation. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

It may state two separate claims as one.

In vivo, attenuating ACAT activity increases levels of 24-hydroxycholesterol and decreases the content of APP and Aβ in the brain of FAD-Tg mice.

“In vivo, attenuating ACAT activity increases the levels of 24-hydroxycholesterol and decreases the content of APP and Aβ in the brain of FAD-Tg mice [73, 74].”

• cites [73] Bryleva, 2010, ACAT1 gene ablation increases 24(S)-hydroxycholesterol content in the brain and ameliorates amyloid pathology in mice with AD, Proc Natl Acad Sci U S A sourceSupported
• cites [74] Murphy, 2013, Acat1 knockdown gene therapy decreases amyloid-beta in a mouse model of Alzheimer's disease, Mol Ther sourceNot checked yet

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

Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

ABCA1 modulates AD pathogenesis, with ABCA1 antagonism increasing and agonism decreasing pathology.

“Over a decade of evidence demonstrates that the cell-surface lipid transporter ATP binding cassette subfamily A member 1 (ABCA1) modulates AD pathogenesis, with ABCA1 antagonism increasing, and agonism decreasing, pathology [48–51].”

• cites [48] Wahrle, 2005, Deletion of Abca1 increases Aβ deposition in the PDAPP transgenic mouse model of Alzheimer disease, J Biol Chem sourcePartly supported

  The Abstract reports that deleting Abca1 increased amyloid deposition and cerebral amyloid angiopathy in PDAPP mice, supporting the claim that reduced ABCA1 activity increases pathology in this model. The Discussion says that increasing ABCA1 might decrease amyloid deposition, but explicitly calls this an untested hypothesis; it does not establish that ABCA1 agonism decreases pathology. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [49] Wahrle, 2008, Overexpression of ABCA1 reduces amyloid deposition in the PDAPP mouse model of Alzheimer disease, J Clin Invest sourceSupported
• cites [50] Wahrle, 2004, ABCA1 is required for normal central nervous system ApoE levels and for lipidation of astrocyte-secreted apoE, J Biol Chem sourcePartly supported

  The Discussion (page not provided) reports that ABCA1 regulates CNS apoE levels and lipidation and suggests that changing ABCA1 function may influence AD-related pathology. The paper does not test ABCA1 antagonism or agonism, or establish the claimed directions of their effects on pathology. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [51] Riddell, 2007, The LXR agonist TO901317 selectively lowers hippocampal abeta42 and improves memory in the Tg2576 mouse model of Alzheimer's disease, Mol Cell Neurosci sourceNot checked — no full text of the source

Probably not a substantial claim. The paper’s conclusions may not depend on it.

Outside lipid rafts, APP is processed at the plasma membrane by α-secretase and -secretases producing sAPP-α via the non-amyloidogenic pathway.

“Outside of lipid rafts, APP is processed at the PM by α-secretase and -secretases producing sAPP-α (non-amyloidogenic pathway) [38–40].”

• cites [38] Hartmann, 2007, Alzheimer's disease: the lipid connection, J Neurochem sourceNot checked — no full text of the source
• cites [39] Ehehalt, 2003, Amyloidogenic processing of the Alzheimer beta-amyloid precursor protein depends on lipid rafts, J Cell Biol sourceSupported
• cites [40] Wang, 2021, Regulation of beta-amyloid production in neurons by astrocyte-derived cholesterol, Proc Natl Acad Sci U S A sourceSupported

It may state two separate claims as one.

AVAS treatment reduces APP processing, reducing the production of Aβ, consistent with previous studies using FAD-Tg mouse models expressing mouse apoE.

“Thus, AVAS treatment reduces APP processing, reducing the production of Aβ, consistent with previous studies using FAD-Tg mouse models expressing m-apoE [76, 105].”

• cites [76] Huttunen, 2010, The acyl-coenzyme A: cholesterol acyltransferase inhibitor CI-1011 reverses diffuse brain amyloid pathology in aged amyloid precursor protein transgenic mice, J Neuropathol Exp Neurol sourcePartly supported

  The Introduction identifies CI-1011 as avasimibe, and the Results report reduced APP cleavage fragments and Aβ levels in treated hAPP transgenic mice (page unavailable). The Discussion suggests that treatment may limit generation of new Aβ, but presents this mechanism as a possibility rather than an established reduction in Aβ production. The passages do not specify whether the mice expressed mouse ApoE. (LLM, conf=0.84)

  Experimental — accuracy not yet measured
• cites [105] Huttunen, 2009, Inhibition of acyl-coenzyme A: cholesterol acyl transferase modulates amyloid precursor protein trafficking in the early secretory pathway, FASEB J sourcePartly supported

  The Results report that ACAT inhibitors reduced mature APP and APP processing products, and describe reduced Aβ in treated transgenic mice. However, the treatments named are CP-113,818 and CI-1011, not AVAS; the Materials and Methods describe the hAPP transgenic mice but do not establish that they expressed mouse apoE. (LLM, conf=0.85)

  Experimental — accuracy not yet measured

Probably not a substantial claim. The paper’s conclusions may not depend on it.

The decrease in endothelial cell disruption and atherosclerotic plaque development was dependent on apoA1 as the plasma cholesterol acceptor.

“This decrease in endothelial cell disruption and atherosclerotic plaque development was dependent on apoA1 as the plasma cholesterol acceptor [57–63].”

• cites [57] Sugimoto, 2004, An inhibitor of acylCoA: cholesterol acyltransferase increases expression of ATP-binding cassette transporter A1 and thereby enhances the ApoA-I-mediated release of cholesterol from macrophages, Biochim Biophys Acta sourceNot checked — no full text of the source
• cites [58] Rodriguez, 2002, Anti-atherogenic effects of the acyl-CoA:cholesterol acyltransferase inhibitor, avasimibe (CI-1011), in cultured primary human macrophages, Atherosclerosis sourceNot checked — no full text of the source
• cites [59] Kellner-Weibel, 2003, Cytotoxic cellular cholesterol is selectively removed by apoA-I via ABCA1, Atherosclerosis sourceNot checked — no full text of the source
• cites [60] Huang, 2001, Sterol efflux mediated by endogenous macrophage ApoE expression is independent of ABCA1, Arterioscler Thromb Vasc Biol sourceNot checked — no full text of the source
• cites [61] An, 2008, Inhibition of acyl-coenzyme A:cholesterol acyltransferase stimulates cholesterol efflux from macrophages and stimulates farnesoid X receptor in hepatocytes, Exp Mol Med sourceCould not verify

  The Introduction and Discussion (page unavailable) discuss cholesterol efflux from macrophages and a possible benefit for atherosclerosis, but do not report a decrease in endothelial disruption or plaque development. The cholesterol-efflux assay describes no apoA1-dependent test; searches for apoA1, cholesterol acceptors, endothelial cells, and plaques found no passage establishing the claimed dependence. (LLM, conf=0.77)

  Experimental — accuracy not yet measured
• cites [62] Yamauchi, 2004, Intracellular cholesterol mobilization involved in the ABCA1/apolipoprotein-mediated assembly of high density lipoprotein in fibroblasts, J Lipid Res sourcePartly supported

  The section on the ACAT-accessible cholesterol pool reports that apoA-I mediates cholesterol release in cells expressing ABCA1. That supports apoA-I’s role in cellular cholesterol removal, but the paper does not report decreased endothelial cell disruption or atherosclerotic plaque development, or show that either outcome depends on apoA-I acting as a plasma cholesterol acceptor. (LLM, conf=0.82)

  Experimental — accuracy not yet measured
• cites [63] Schmitz, 1985, Regulation of high density lipoprotein receptors in cultured macrophages: role of acyl-CoA:cholesterol acyltransferase, EMBO J sourceNot checked — no full text of the source

It may state two separate claims as one.

In this study, AVAS did not produce an overall improvement during Morris water maze acquisition trials; latency to platform on day 1 was lower in AVAS- versus vehicle control-treated mice, suggesting enhanced learning on that day only, and latency was not significantly different between groups for days 2-5.

“While we did not see an overall improvement during MWM acquisition trials with AVAS treatment, the latency to platform on day 1 was lower in AVAS- vs. VC-treated mice, suggesting that the AVAS treatment enhanced learning on that day only; the latency to platform was not significantly different between VC- and AVAS-treated mice for days 2–5 (Fig. 3b), an effect observed with other AVAS-treated FAD-”

• cites [75] Hutter-Paier, 2004, The ACAT inhibitor CP-113,818 markedly reduces amyloid pathology in a mouse model of Alzheimer's disease, Neuron sourcePartly supported

  Split into 2 atoms — 0 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 1 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured

It may state two separate claims as one.

Mouse apoE and human apoE differ in both structure and functions ranging from lipid binding and transport to effects on AD pathology.

“M-apoE and h-apoE differ in both structure [119] and functions ranging from lipid binding and transport, to effects on AD pathology [22, 84, 85, 119, 120].”

• cites [119] Nguyen, 2014, Influence of domain stability on the properties of human apolipoprotein E3 and E4 and mouse apolipoprotein E, Biochemistry sourcePartly supported

  The Discussion section, “Interaction of Mouse ApoE Variants with Lipids,” reports differences between mouse and human apoE in domain behavior and lipid- and lipoprotein-binding properties. The paper mentions Alzheimer’s disease risk for human apoE isoforms, but does not compare mouse and human apoE effects on AD pathology. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [22] Fagan, 1999, Unique lipoproteins secreted by primary astrocytes from wild type, apoE (-/-), and human apoE transgenic mice, J Biol Chem sourcePartly supported

  The discussion describes structural differences between mouse and human apoE, and the Results report different lipid-to-apoE ratios in particles containing each form. The paper discusses findings on amyloid-β deposition but says the particles’ differences may differentially affect deposition; it does not establish a direct mouse-versus-human difference in effects on AD pathology. (LLM, conf=0.85)

  Experimental — accuracy not yet measured
• cites [84] Liao, 2015, Murine versus human apolipoprotein E4: differential facilitation of and co-localization in cerebral amyloid angiopathy and amyloid plaques in APP transgenic mouse models, Acta Neuropathol Commun sourcePartly supported

  The Introduction reports 70% sequence homology between human apoE4 and mouse apoE, and the Discussion reports that mouse apoE promoted plaques while human apoE4 promoted cerebral amyloid angiopathy. These passages support structural and AD-pathology differences, but our copy does not establish a difference in lipid binding or transport between them (searched: “lipid binding and transport” and “lipid transport lipid binding difference mouse human apoE”). (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [85] Fagan, 2002, Human and murine ApoE markedly alters A beta metabolism before and after plaque formation in a mouse model of Alzheimer's disease, Neurobiol Dis sourceNot checked — no full text of the source
• cites [120] Hudry, 2013, Gene transfer of human apoe isoforms results in differential modulation of amyloid deposition and neurotoxicity in mouse brain, Sci Transl Med sourcePartly supported

  The Discussion (page unavailable) reports that replacing murine Apoe with human APOE isoforms modifies amyloid deposits, and describes effects of the human isoforms on AD-related pathology. It does not establish a mouse-versus-human structural difference or compare their lipid binding and transport functions. (LLM, conf=0.82)

  Experimental — accuracy not yet measured

Probably not a substantial claim. The paper’s conclusions may not depend on it.

AVAS is currently being proposed as an anti-cancer agent.

“is currently being proposed as an anti-cancer agent [137]”

• cites [137] Zabielska, 2019, Acyl-coenzyme a: cholesterol acyltransferase inhibition in cancer treatment, Anticancer Res sourceSupported

It may state two separate claims as one.

ACAT inhibition significantly reduces palmitoylated APP in lipid rafts, thus reducing APP processing.

“ACAT inhibition significantly reduces palmitoylated APP in lipid rafts, thus reducing APP processing [75–77, 105, 131].”

• cites [75] Hutter-Paier, 2004, The ACAT inhibitor CP-113,818 markedly reduces amyloid pathology in a mouse model of Alzheimer's disease, Neuron sourcePartly supported

  The indexed passage on APP processing (labeled Abstract; page unavailable) reports that CP-113,818 significantly reduced total secreted APP in mouse brain, supporting reduced APP processing. It does not report a reduction in palmitoylated APP in lipid rafts or establish that mechanism as the cause of reduced processing. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [76] Huttunen, 2010, The acyl-coenzyme A: cholesterol acyltransferase inhibitor CI-1011 reverses diffuse brain amyloid pathology in aged amyloid precursor protein transgenic mice, J Neuropathol Exp Neurol sourcePartly supported

  The Results section ('ACAT Inhibition by CI-1011 Reduces APP Processing and Aβ Generation in Cells'; page unavailable) reports reduced APP C-terminal fragments and secreted Aβ after CI-1011 treatment, supporting reduced APP processing. The paper does not report a significant reduction in palmitoylated APP in lipid rafts or establish that as the mechanism; its Discussion instead proposes an effect on APP trafficking and maturation. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [77] Bhattacharyya, 2010, ACAT inhibition and amyloid beta reduction, Biochim Biophys Acta sourcePartly supported

  Section 2.2 reports that ACAT inhibitors reduce APP processing, but says the cellular mechanism remains largely unknown. It does not report a significant reduction of palmitoylated APP in lipid rafts or establish that change as the cause of reduced processing; its discussion of lipid rafts is speculative. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [105] Huttunen, 2009, Inhibition of acyl-coenzyme A: cholesterol acyl transferase modulates amyloid precursor protein trafficking in the early secretory pathway, FASEB J sourcePartly supported

  The Results report reduced mature APP and APP processing fragments after ACAT inhibition, and the Discussion links reduced APP maturation to reduced Aβ generation (page numbers unavailable). The paper does not report a reduction in palmitoylated APP in lipid rafts or establish that as the mechanism. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [131] Bhattacharyya, 2013, Palmitoylation of amyloid precursor protein regulates amyloidogenic processing in lipid rafts, J Neurosci sourcePartly supported

  In the Results section (Fig. 9F), ACAT1 inhibition reduced lipid raft-bound palAPP by 76.15 ± 8.9%, alongside reduced APP cleavage-product levels. The paper says the palAPP reduction *likely* resulted in decreased amyloidogenic APP processing; the claim presents that causal link as established. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

It may state two separate claims as one.

AVAS reduces soluble oligomeric Aβ, soluble and insoluble Aβ42, Aβ deposition, amyloid deposition, and neuroinflammation in a FAD-Tg mouse expressing human apoE4, not mouse apoE, and without modifying the lipidation state of apoE particles.

“Specifically, we show a reduction in soluble oAβ, soluble and insoluble Aβ42 levels, Aβ deposition, amyloid deposition, and neuroinflammation (Figs. 5 and 6) [75, 76]. Remarkably, this occurs in a FAD-Tg-mouse expressing h-apoE4, not m-apoE, and without modifying the lipidation state of apoE-particles (Fig. 7).”

• cites [75] Hutter-Paier, 2004, The ACAT inhibitor CP-113,818 markedly reduces amyloid pathology in a mouse model of Alzheimer's disease, Neuron sourcePartly supported

  The Abstract (page unavailable) reports reduced soluble Aβ42, insoluble Aβ, and amyloid plaques in transgenic mice, supporting those outcomes in the cited sentence. But the treatment tested was CP-113,818, not AVAS, and our copy does not report reductions in soluble oligomeric Aβ or neuroinflammation. The human apoE4 and apoE-particle lipidation details are presented as the citing authors’ own Figure 7 finding, rather than a finding attributed to this source. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [76] Huttunen, 2010, The acyl-coenzyme A: cholesterol acyltransferase inhibitor CI-1011 reverses diffuse brain amyloid pathology in aged amyloid precursor protein transgenic mice, J Neuropathol Exp Neurol sourcePartly supported

  The Results report that CI-1011 (avasimibe) reduced diffuse amyloid deposits and SDS-extractable Aβ42 in aged hAPP mice. They do not report a reduction in soluble oligomeric Aβ, and the change in formic-acid-extracted Aβ42 was not significant. The paper reports reduced cortical astrogliosis but increased hippocampal microglial activation, rather than an unqualified reduction in neuroinflammation. The h-apoE4 and unchanged apoE-particle-lipidation statements appear in the citing paper’s next sentence, to which this citation is not attached; this source does not establish them. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Probably not a substantial claim.

Cholesterol mobilization by ACAT inhibition in microglia can promote autophagy and lysosomal biogenesis, leading to increased clearance of oAβ.

“cholesterol mobilization by ACAT inhibition in microglia can promote autophagy and lysosomal biogenesis, leading to an increased clearance of oAβ [69–71, 77].”

• cites [69] Chang, 2021, Blocking cholesterol storage to treat Alzheimer's disease, Explor Neuroprotective Ther sourceSupported
• cites [70] Rubio, 2019, ACAT1 as a therapeutic target and its genetic relationship with Alzheimer's disease, Curr Alzheimer Res sourceNot checked — no full text of the source
• cites [71] Shibuya, 2014, Inhibiting ACAT1/SOAT1 in microglia stimulates autophagy-mediated lysosomal proteolysis and increases Abeta1-42 clearance, J Neurosci sourceSupported
• cites [77] Bhattacharyya, 2010, ACAT inhibition and amyloid beta reduction, Biochim Biophys Acta sourcePartly supported

  Sections 2.1 and 2.2 report that ACAT inhibition reduces Aβ levels and affects cellular cholesterol distribution. But section 2.1 says an effect on Aβ clearance through unidentified mechanisms has not been excluded; it does not establish increased clearance. The cited passages do not support the specific mechanism involving microglial autophagy, lysosomal biogenesis, or clearance of oligomeric Aβ. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Probably not a substantial claim. The paper’s conclusions may not depend on it.

AVAS has previously been administered to FAD-Tg mice by subcutaneous pellet.

“AVAS has previously been administered to FAD-Tg mice by subcutaneous pellet [76].”

• cites [76] Huttunen, 2010, The acyl-coenzyme A: cholesterol acyltransferase inhibitor CI-1011 reverses diffuse brain amyloid pathology in aged amyloid precursor protein transgenic mice, J Neuropathol Exp Neurol sourceSupported

It may state two separate claims as one.

In this study, AVAS treatment reduced both amyloidogenic and non-amyloidogenic processing of APP, with no change in total APP.

“we did see a reduction in both amyloidogenic and non-amyloidogenic processing of APP, with no change in total APP (Fig. 8)”

• cites [75] Hutter-Paier, 2004, The ACAT inhibitor CP-113,818 markedly reduces amyloid pathology in a mouse model of Alzheimer's disease, Neuron sourcePartly supported

  The APP-processing passage (listed as Abstract; page unavailable) reports unchanged full-length APP and reduced secreted α- and β-secretase products after CP-113,818 treatment. The paper identifies avasimibe as a different ACAT inhibitor; it does not report these APP-processing results for AVAS. In nontransgenic mice, the decrease in sAPPα was not statistically significant. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [76] Huttunen, 2010, The acyl-coenzyme A: cholesterol acyltransferase inhibitor CI-1011 reverses diffuse brain amyloid pathology in aged amyloid precursor protein transgenic mice, J Neuropathol Exp Neurol sourcePartly supported

  The Results sections report that CI-1011 (avasimibe) reduced both APP-C99 and APP-C83, supporting reduced amyloidogenic and non-amyloidogenic processing. The copy does not establish that total APP was unchanged; the Discussion instead says treatment appears to affect mature APP. Page numbers are unavailable in the indexed copy. (LLM, conf=0.85)

  Experimental — accuracy not yet measured

Probably not a substantial claim. The paper’s conclusions may not depend on it.

Previous studies demonstrate that ACAT inhibition improves Morris water maze learning.

“With MWM, previous studies demonstrate that ACAT inhibition improves MWM learning [75].”

• cites [75] Hutter-Paier, 2004, The ACAT inhibitor CP-113,818 markedly reduces amyloid pathology in a mouse model of Alzheimer's disease, Neuron sourceSupported

It may state two separate claims as one.

Interventions that promote cholesterol-regulating processes reduce Aβ pathology, including increasing ER-free cholesterol via inhibiting cholesteryl ester formation by ACAT1 inhibition.

“Interestingly, interventions that promote these processes reduce Aβ pathology, including increasing ER-FC via inhibiting CE formation by ACAT1 inhibition [74, 75, 103, 111].”

• cites [74] Murphy, 2013, Acat1 knockdown gene therapy decreases amyloid-beta in a mouse model of Alzheimer's disease, Mol Ther sourcePartly supported

  The Introduction states that ACAT enzymes form cholesteryl esters, and the Results (‘Testing of AAV gene therapy effects on AD mice’; page unavailable) report significantly lower Aβ42 after Acat1 knockdown in AD mice. The paper does not establish that this treatment increased ER-free cholesterol or that such an increase caused the Aβ reduction; the Discussion says the molecular mechanism remains incompletely understood. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [75] Hutter-Paier, 2004, The ACAT inhibitor CP-113,818 markedly reduces amyloid pathology in a mouse model of Alzheimer's disease, Neuron sourcePartly supported

  The Abstract reports that ACAT inhibitor CP-113,818 reduced brain cholesteryl esters and amyloid pathology in transgenic mice. The Introduction identifies ACAT as an endoplasmic-reticulum enzyme that converts cholesterol to cholesteryl esters, but the paper does not establish an increase in ER-free cholesterol; it reports no change in overall brain free-cholesterol levels. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [103] Zhu, 2018, In vitro exploration of ACAT contributions to lipid droplet formation during adipogenesis, J Lipid Res sourcePartly supported

  The Discussion reports that ACAT1 inhibition suppresses cholesteryl ester synthesis in neurons and that cholesteryl ester forms on the ER membrane. It does not report increased ER free cholesterol or reduced Aβ pathology; its own adipocyte results instead show decreased intracellular free cholesterol. The References list a separate work on ACAT inhibition and amyloid-beta reduction, but that title does not establish the claimed outcome in this paper. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [111] Huttunen, 2008, ACAT as a drug target for Alzheimer’s disease, Neurodegener Dis sourcePartly supported

  The paper’s discussion of ACAT inhibition (pp. 212–214) says ACAT is an ER-resident enzyme that converts free cholesterol to cholesteryl esters, and reports that inhibiting ACAT reduces Aβ generation and amyloid pathology in cell and animal models. It says inhibition changes the distribution of free and esterified cholesterol, but does not specifically establish an increase in ER free cholesterol. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

Lipid droplets are composed of neutral lipids, primarily cholesteryl esters and triacylglycerol, so the data are consistent with previous findings that inhibition of ACAT reduces brain cholesteryl ester levels.

“LD are composed of neutral lipids, primarily CE and TAG, thus our data are consistent with previous findings that inhibition of ACAT reduces brain CE levels [75, 76].”

• cites [75] Hutter-Paier, 2004, The ACAT inhibitor CP-113,818 markedly reduces amyloid pathology in a mouse model of Alzheimer's disease, Neuron sourceSupported
• cites [76] Huttunen, 2010, The acyl-coenzyme A: cholesterol acyltransferase inhibitor CI-1011 reverses diffuse brain amyloid pathology in aged amyloid precursor protein transgenic mice, J Neuropathol Exp Neurol sourceSupported

Probably not a substantial claim. The paper’s conclusions may not depend on it.

APOE4 astrocytes chronically exposed to fatty acids produce TAG-rich apoE-particles rather than cholesterol-rich particles.

“APOE4 astrocytes chronically exposed to fatty acids (a possible mimic for aged astrocytes rich in LD) produce TAG-rich apoE-particles rather than cholesterol-rich particles [130].”

• cites [130] Lindner, 2022, Isoform- and cell-state-specific lipidation of ApoE in astrocytes, Cell Rep sourceSupported

It may state two separate claims as one.

In vitro, ACAT inhibition has cell-specific effects on ABCA1 and cholesterol efflux: it increases ABCA1 levels and cholesterol efflux in cholesterol-loaded macrophages, decreases ABCA1 levels in adipocytes, and decreases ABCA1 levels, lipid catabolism, and cholesterol efflux in APOE4 microglia.

“In vitro, ACAT inhibition increases ABCA1 levels and cholesterol efflux of cholesterol-loaded macrophages, while decreasing ABCA1 levels in adipocytes [125] and decreasing ABCA1 levels, lipid catabolism, and cholesterol efflux in APOE4 microglia [126–128]”

• cites [125] Reverter, 2011, Cholesterol transport from late endosomes to the Golgi regulates t-SNARE trafficking, assembly, and function, Mol Biol Cell sourceCould not verify

  Split into 3 atoms — 0 confirmed, 0 partially supported, 0 disputed, 0 misattributed, 3 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured
• cites [126] Lim, 2013, Effects of a high-fat, high-cholesterol diet on brain lipid profiles in apolipoprotein E epsilon3 and epsilon4 knock-in mice, Neurobiol Aging sourceNot checked — no full text of the source
• cites [127] Mahley, 2016, Central nervous system lipoproteins: ApoE and regulation of cholesterol metabolism, Arterioscler Thromb Vasc Biol sourcePartly supported

  Split into 3 atoms — 0 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 2 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured
• cites [128] Zhao, 2017, APOE epsilon4/epsilon4 diminishes neurotrophic function of human iPSC-derived astrocytes, Hum Mol Genet sourceNot checked — no full text of the source

Probably not a substantial claim.

The sequential protein extraction method was optimized to extract lipoprotein-associated apoE to the TBSX fraction using 1% Triton X-100, allowing intact lipoproteins to extract in this fraction.

“The protocol for this sequential protein extraction method was originally optimized to extract lipoprotein-associated apoE to the TBSX fraction by using 1%Triton X-100, a non-ionic detergent that is less stringent compared to SDS or Tween 20, allowing the intact lipoproteins to extract in this fraction [99].”

• cites [99] Youmans, 2011, Amyloid-beta42 alters apolipoprotein E solubility in brains of mice with five familial AD mutations, J Neurosci Methods sourcePartly supported

  Methods §2.3 describes the TBSX extraction using 1% Triton X-100, and the Discussion says this method extracts lipoprotein-associated apoE primarily into TBSX in wild-type mice. The paper does not say that lipoprotein particles remain intact in that fraction or establish the claimed comparison with SDS or Tween 20. No page numbers are available in our copy. (LLM, conf=0.87)

  Experimental — accuracy not yet measured

It may state two separate claims as one.

The brain/blood bioavailability ratio of AVAS was low (approximately 0.01), but measured brain concentrations were above the desired pharmacodynamic threshold at 3 h and 6 h.

“Although the brain/blood bioavailability ratio was low (≈0.01), measured brain concentrations were above the desired PD threshold at 3 h and 6 h (Fig. 1d).”

It may state two separate claims as one.

Both Aβ and amyloid deposition are significantly reduced by approximately 50% in the cortex of AVAS versus vehicle control-treated mice.

“Both Aβ (Fig. 5b) and amyloid deposition (Fig. 5c) are significantly reduced by ~ 50% in the CX of AVAS vs. VC-treated mice.”

It may state two separate claims as one.

In AVAS-treated mice compared to vehicle control, body weights decreased significantly but to a maximum of 5% from week 2-6, with no significant difference in weeks 7-8.

“In the AVAS-treated mice compared to VC, body weights decreased significantly but to a maximum of 5% from week 2–6, with no significant difference in weeks 7–8 (Fig. 2b).”

It may state two separate claims as one.

Primary glial cells treated for 24 h with non-cytotoxic AVAS concentrations (0.05-1.0 μg/ml) induced a 50% increase at 0.05 μg/ml and a 2-fold increase at 0.1 and 0.5 μg/ml in apoE levels in the media, with no treatment effect at 6 h.

“Primary glial cells treated for 24 h with non-cytotoxic concentrations of AVAS (0.05–1.0 μg/ml) induced a 50% (0.05 μg/ml) and 2-fold (0.1 and 0.5 μg/ml) increase in apoE levels in the media, with no treatment-effect at 6 h (Fig. 1b).”

It may state two separate claims as one.

Levels of the post-synaptic proteins PSD95 and drebrin are significantly increased 50% and 2-fold, respectively, with AVAS treatment compared to vehicle control.

“Levels of the post-synaptic proteins PSD95 and drebrin are significantly increased 50% and 2-fold, respectively, with AVAS treatment compared to VC.”

It may state two separate claims as one.

AVAS at 2.5-10.0 μg/ml caused a small but significant reduction in viability at 6 h, and at 24 h viability dropped to approximately 25%.

“From 2.5–10.0 μg/ml, there was a small but significant reduction in viability for the 6 h treatment while viability for 24 h treatment dropped to ~ 25%.”

It may state two separate claims as one.

Both astrogliosis and microgliosis are significantly reduced by 50% in the cortex in AVAS versus vehicle control-treated mice.

“Both astrogliosis (Fig. 6b) and microgliosis (Fig. 6c) are significantly reduced by 50% in the CX in AVAS vs. VC treated mice.”

It may state two separate claims as one.

ACAT inhibition by AVAS can promote an increase in extracellular apoE levels in glial cells and/or promote apoE stability in the media of glial cells.

“These results demonstrate that ACAT inhibition by AVAS can promote an increase in extracellular apoE levels in glial cells and/or promote apoE stability in the media of glial cells.”

Probably not a substantial claim. It may state two separate claims as one.

The amount of cholesterol in neuronal lipid rafts is regulated by astrocytic apoE-lipoproteins, promoting or limiting APP processing by β- and γ-secretases and hence Aβ production.

“Recent data suggest that the amount of cholesterol in neuronal lipid rafts is regulated by astrocytic apoE-lipoproteins promoting or limiting APP processing by β- and γ-secretases, and hence the amount of Aβ production [40].”

• cites [40] Wang, 2021, Regulation of beta-amyloid production in neurons by astrocyte-derived cholesterol, Proc Natl Acad Sci U S A sourceNot checked yet

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

Probably not a substantial claim. The paper’s conclusions may not depend on it.

APOE4 is the greatest genetic risk factor for Alzheimer's disease compared to the common APOE3.

“APOE4, encoding apolipoprotein E4 (apoE4), is the greatest genetic risk factor for Alzheimer’s disease (AD), compared to the common APOE3.”

• Also stated in: § Introduction · plain

  “The greatest genetic risk factor for sporadic AD is inheritance of the ε4 allele of the human (h)-APOE gene compared to the common ε3 and the neuroprotective but rare ε2 allele [10].”

• cites [10] Alzheimer's Association, 2023, 2023 Alzheimer's disease facts and figures, Alzheimers DementNot checked yet

It may state two separate claims as one.

The AVAS-induced reduction in Aβ via reduced APP processing was sufficient to reduce AD pathology, as apoE4-lipoproteins remained poorly lipidated.

“This suggests that the AVAS-induced reduction in Aβ via reduced APP processing was sufficient to reduce AD pathology, as apoE4-lipoproteins remained poorly lipidated.”

It may state two separate claims as one.

ACAT catalyzes intracellular cholesterol esterification, reducing the ER-free cholesterol pool by forming cholesteryl esters stored as lipid droplets.

“The endoplasmic reticulum (ER)-resident ACAT catalyzes intracellular cholesterol esterification, critical in this context as it reduces the ER-free cholesterol (ER-FC) pool by the formation of CE stored as LD [56].”

• cites [56] Chang, 2009, Acyl-coenzyme A:cholesterol acyltransferases, Am J Physiol Endocrinol Metab sourceNot checked — no full text of the source

It may state two separate claims as one.

AVAS exhibited both target engagement and efficacy, as well as inhibition of Aβ deposition and solubility, and neuroinflammation.

“Thus, AVAS exhibited both target engagement and efficacy, as well as inhibition of Aβ deposition and solubility, and neuroinflammation.”

Probably not a substantial claim.

Severe hepatomegaly is associated with RXR agonism due to induction of hepatic lipogenic pathways by SREBP1C.

“However, severe hepatomegaly is associated with RXR agonism due to an induction of hepatic lipogenic pathways by sterol-regulatory element-binding protein 1C (SREBP1C) [52, 53].”

• cites [52] Tai, 2014, Amyloid-β pathology and APOE genotype modulate retinoid X receptor agonist activity in vivo, J Biol Chem sourceNot checked yet

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

• cites [53] Koster, 2017, Rexinoids as therapeutics for Alzheimer's disease: role of APOE, Curr Top Med Chem sourceNot checked — no full text of the source

Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

ACAT exists as two isoforms: ACAT1 is distributed ubiquitously, and ACAT2 is expressed primarily by enterocytes and hepatocytes.

“ACAT exists as two isoforms, ACAT1 is distributed ubiquitously, and ACAT2 is expressed primarily by enterocytes and hepatocytes [54, 55].”

• cites [54] Meiner, 1997, Tissue expression studies on the mouse acyl-CoA: cholesterol acyltransferase gene (Acact): findings supporting the existence of multiple cholesterol esterification enzymes in mice, J Lipid Res sourceNot checked — no full text of the source
• cites [55] Cases, 1998, ACAT-2, a second mammalian acyl-CoA:cholesterol acyltransferase. Its cloning, expression, and characterization, J Biol Chem sourceNot checked — no full text of the source

It may state two separate claims as one.

The AVAS-induced improvement in Morris water maze memory is consistent with the increase in post-synaptic proteins, indicating that AVAS exhibits surrogate efficacy.

“Thus, the AVAS-induced improvement in MWM memory is consistent with the increase in post-synaptic proteins, indications that AVAS exhibits surrogate efficacy.”

Probably not a substantial claim. The paper’s conclusions may not depend on it.

AVAS failed phase III clinical trials for cardiovascular efficacy.

“Although AVAS failed phase III clinical trials for cardiovascular efficacy [101]”

• cites [101] Tardif, 2004, Effects of the acyl coenzyme A:cholesterol acyltransferase inhibitor avasimibe on human atherosclerotic lesions, Circulation sourceNot checked — no full text of the source

Probably not a substantial claim. The paper’s conclusions may not depend on it.

In pre-clinical studies, regulation of brain cholesterol levels through different strategies has successfully reduced Aβ pathology.

“In pre-clinical studies, regulation of brain cholesterol levels through different strategies has successfully reduced Aβ pathology.”

Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

AVAS-induced inhibition of peripheral cholesterol esterification was originally a therapeutic target for atherosclerosis as it facilitated cholesterol efflux to apoA1-HDL in plasma, reducing development of foam cells from macrophage uptake of oxidized LDL.

“AVAS-induced inhibition of peripheral cholesterol esterification was originally a therapeutic target for atherosclerosis as it facilitated cholesterol efflux to apoA1-HDL in plasma, thus reducing the development of foam cells from macrophage uptake of oxidized LDL.”

It may state two separate claims as one.

Humans express three primary apoE isoforms that vary at only two of the 299 amino acids; mice express one form of apoE, the same as apoE4 at residues Arg112/Arg158, although homology between mouse and human apoE is only 70%.

“While humans express three primary isoforms that vary at only two of the 299 amino acids, mice express one form of apoE, the same as apoE4 at residues Arg112/Arg158, although the homology between m- and h-apoE is only 70% [118].”

• cites [118] Rajavashisth, 1985, Evolution of apolipoprotein E: mouse sequence and evidence for an 11-nucleotide ancestral unit, Proc Natl Acad Sci U S A sourceNot checked — no full text of the source

Probably not a substantial claim. The paper’s conclusions may not depend on it.

The study did not measure APP palmitoylation or APP localization to lipid rafts.

“Although we did not measure APP palmitoylation or APP localization to lipid rafts”

Probably not a substantial claim. The paper’s conclusions may not depend on it.

A 24 h interval between the last training trial and the probe trial allows reference memory to be tested, independent of the short-term memory of the last training session.

“A 24 h interval between the last training trial and the probe trial allows reference memory to be tested, independent of the short-term memory of the last training session [94].”

• cites [94] Baldi, 2005, Spatial navigation in the Morris water maze: working and long lasting reference memories, Neurosci Lett sourceNot checked — no full text of the source

Probably not a substantial claim. The paper’s conclusions may not depend on it.

Male E4FAD mice were randomized into two treatment groups.

“Male E4FAD mice (n = 20) were randomized into 2 treatment groups (n = 10): vehicle control (VC) and AVAS (AVAS).”

It may state two separate claims as one.

AD pathology is reduced by ACAT inhibition, consistent with previous studies.

“AD pathology is reduced by ACAT inhibition, consistent with previous studies.”

It may state two separate claims as one.

The AVAS-induced changes are not sufficient to increase lipid efflux and modify apoE4-particles.

“However, these AVAS-induced changes are not sufficient to increase lipid efflux and modify apoE4-particles.”

It may state two separate claims as one.

AVAS-induced redistribution of cholesterol within the cell is sufficient to modify LDs and lipid raft formation/distribution, resulting in increased synaptic function and reduced APP processing.

“Overall, our results suggest that the AVAS-induced redistribution of cholesterol within in the cell is sufficient to modify LDs and lipid raft formation/distribution, resulting in increased synaptic function and reduced APP processing.”