APOE4-drives brain bioenergetic shifts independent of Alzheimer’s disease pathology

May combine two claims.

APOE4 alters blood-brain barrier integrity, influencing metabolite exchange between the periphery and CNS, including reduced brain glucose and lipid transport.

“APOE4 alters the integrity of the blood brain barrier (BBB) influencing metabolite exchange between the periphery and the central nervous system (CNS) including reduced brain glucose and lipid transport [17][18][19] .”

• cites [17] Bell, R. D. et al. Apolipoprotein E controls cerebrovascular integrity via cyclophilin A. Nature 485, 512–516 (2012). sourcePartly supported

  The paper reports that APOE4-expressing mice have a leaky blood–brain barrier and increased entry of blood-derived proteins into the brain (Abstract; page not provided). This supports altered barrier integrity and exchange across it, but the paper does not report reduced brain glucose or lipid transport. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [18] Liu, C. C. et al. Peripheral apoE4 enhances Alzheimer’s pathology and impairs cognition by compromising cerebrovascular function. Nat. Neurosci. 25, 1020–1033 (2022). sourcePartly supported

  The Results section (“Peripheral apoE4 impairs cerebrovascular functions”; page unavailable) reports increased BBB leakage and reduced tight-junction markers in mice expressing liver-derived apoE4. This supports altered barrier integrity and movement of blood-derived proteins into the brain. The paper does not establish reduced transport of glucose or lipids into the brain; its discussion of metabolism and lipid-related genes does not measure that transport. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [19] Montagne, A. et al. APOE4 leads to blood-brain barrier dysfunction predicting cognitive decline. Nature 581, 71–76 (2020). sourcePartly supported

  The Introductory Paragraph reports greater blood–brain barrier permeability in APOE4 carriers, supporting the claim that APOE4 alters barrier integrity. It does not establish reduced brain glucose or lipid transport, or measure metabolite exchange between the periphery and CNS; searches for those transport claims found no supporting passage in our copy. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim.

Prior lipidomic, metabolomic, and magnetic resonance spectroscopy studies in mouse models show that APOE genetic variation modifies brain metabolite levels.

“Prior lipidomic, metabolomic, and magnetic resonance spectroscopy (MRS) studies in mouse models show that genetic variation in APOE modify brain metabolite levels [20][21][22][23] .”

• cites [20] Johnson, L. A., Torres, E. R., Impey, S., Stevens, J. F. & Raber, J. Apolipoprotein E4 and Insulin Resistance Interact to Impair Cognition and Alter the Epigenome and Metabolome. Sci. Rep. 7, 43701 (2017). sourceSupported
• cites [21] Area-Gomez, E. et al. APOE4 is Associated with Differential Regional Vulnerability to Bioenergetic Deficits in Aged APOE Mice. Sci. Rep. 10, 4277 (2020). sourceSupported
• cites [22] Lee, S. et al. APOE modulates microglial immunometabolism in response to age, amyloid pathology, and inflammatory challenge. Cell Rep. 42, 112196 (2023). sourcePartly supported

  The Results section on MALDI mass spectrometry imaging reports APOE-associated changes in lipid concentrations across mouse brain regions, and the microglia Results section reports APOE-dependent metabolite differences. These findings support the lipidomic and metabolomic parts of the sentence, but our copy does not report magnetic resonance spectroscopy findings. (LLM, conf=0.84)

  Experimental — accuracy not yet measured
• cites [23] Miranda, A. M. et al. Effects of APOE4 allelic dosage on lipidomic signatures in the entorhinal cortex of aged mice. Transl. Psychiatry 12, 129 (2022). sourceSupported

May combine two claims.

APOE4 is associated with increased Aβ deposition, but its influence on metabolic pathways may contribute to AD risk independently of Aβ.

“As such, APOE4 is associated with increased Aβ deposition but accumulating evidence suggests that its influence on metabolic pathways may contribute to disease risk independently of Aβ [3][4][5][6][7] .”

• cites [3] Reiman, E. M. et al. Declining brain activity in cognitively normal apolipoprotein E epsilon 4 heterozygotes: A foundation for using positron emission tomography to efficiently test treatments to prevent Alzheimer’s disease. Proc. Natl. Acad. Sci. USA. 98, 3334–3339 (2001). sourceNot checked — no full text of the source
• cites [4] Reiman, E. M. et al. Preclinical evidence of Alzheimer’s disease in persons homozygous for the epsilon 4 allele for apolipoprotein E. N. Engl. J. Med 334, 752–758 (1996). sourcePartly supported

  The Discussion (page not provided) associates the ε4 allele with faster β-amyloid deposition, and the Abstract reports reduced brain glucose metabolism in cognitively normal ε4 homozygotes. The paper does not establish that the metabolic difference contributes to Alzheimer’s disease risk independently of amyloid deposition. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [5] Wolf, A. B. et al. Apolipoprotein E as a beta-amyloid-independent factor in Alzheimer’s disease. Alzheimers Res Ther. 5, 38 (2013). sourcePartly supported

  The Review’s Introduction describes APOE4-related changes in lipid metabolism and cerebral energy metabolism, including effects reported without a contribution from fibrillar Aβ; this supports the hedged claim that metabolic pathways may contribute independently to AD risk. It also links APOE4 to impaired Aβ clearance, but the passages do not explicitly establish increased Aβ deposition in APOE4 carriers. (LLM, conf=0.82)

  Experimental — accuracy not yet measured
• cites [6] Brandon, J. A., Farmer, B. C., Williams, H. C. & Johnson, L. A. APOE and Alzheimer’s Disease: Neuroimaging of Metabolic and Cerebrovascular Dysfunction. Front Aging Neurosci. 10, 180 (2018). sourcePartly supported

  The section on cerebral blood flow and cerebral amyloid angiopathy links APOE4 to amyloid deposition, while the Abstract and Brain Lipid Metabolism section describe APOE4-associated metabolic impairments that may contribute to AD. The paper does not establish that this metabolic contribution to disease risk is independent of Aβ; the Brain Lipid Metabolism section also discusses possible interactions with Aβ. (LLM, conf=0.84)

  Experimental — accuracy not yet measured
• cites [7] Jagust, W. J. & Landau, S. M. Alzheimer’s Disease Neuroimaging I. Apolipoprotein E, not fibrillar beta-amyloid, reduces cerebral glucose metabolism in normal aging. J. Neurosci. 32, 18227–18233 (2012). sourceSupported

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

APOE genetic variation is a major risk factor for late-onset Alzheimer's disease, with the ε2 allele lowering risk, ε3 neutral, and ε4 increasing risk.

“Genetic variation in apolipoprotein E (APOE) is a major risk factor for late onset AD (LOAD) where the ε2 allele lowers risk, the ε3 allele affords neutral risk, and the ε4 allele increases risk 2 .”

• cites [2] Yang, L. G., March, Z. M., Stephenson, R. A. & Narayan, P. S. Apolipoprotein E in lipid metabolism and neurodegenerative disease. Trends Endocrinol. Metab. 34, 430–445 (2023). sourceSupported

May combine two claims.

Postmortem AD brain from APOE4 carriers displays mitochondrial dysfunction and increased oxidative stress markers.

“Work from our group and others show that postmortem AD brain from APOE4 carriers display mitochondrial dysfunction and increased oxidative stress markers 15,16 .”

• cites [15] Yin, J. et al. Effect of ApoE isoforms on mitochondria in Alzheimer disease. Neurology 94, e2404–e2411 (2020). sourcePartly supported

  The Results report impaired mitochondrial structure and function associated with APOE-ε4 in postmortem brain tissue. But the sample included cognitively normal people and people with mild cognitive impairment as well as AD, rather than only AD brains. The measured oxidative-stress-related proteins, SOD2 and Foxo3a, were lower—not increased—in carriers; the Discussion describes them as downregulated antioxidative-stress proteins. (LLM, conf=0.89)

  Experimental — accuracy not yet measured
• cites [16] Troutwine, B. R. et al. Mitochondrial function and Abeta in Alzheimer’s disease postmortem brain. Neurobiol. Dis. 171, 105781 (2022). sourcePartly supported

  The Results report mitochondrial respiratory-chain deficits in postmortem brain from APOE ε4 carriers, but the carrier analysis combined AD and non-demented subjects; it did not establish the finding specifically within AD carriers. The Introduction mentions increased reactive oxygen species in AD research generally, but the paper does not report increased oxidative-stress markers in postmortem AD brain from APOE ε4 carriers. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

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

DAA pathway phenotypes are associated with AD.

“APOE4 derived iAstrocytes showed increased inflammatory protein expression including those associated with DAA pathway phenotypes in AD [42][43][44][45] .”

• cites [42] Kim, H. et al. Reactive astrocytes transduce inflammation in a blood-brain barrier model through a TNF-STAT3 signaling axis and secretion of alpha 1-antichymotrypsin. Nat. Commun. 13, 6581 (2022). sourcePartly supported

  The Results section reports higher levels of inflammatory reactive astrocyte markers in the AD cohort than in age-matched asymptomatic controls. The paper also lists a study titled “Disease-associated astrocytes in Alzheimer’s disease and aging” in its References, but the passages do not identify the measured cells or proteins as a DAA pathway phenotype. (LLM, conf=0.79)

  Experimental — accuracy not yet measured
• cites [43] Zhao, Q. et al. The origins and dynamic changes of C3- and S100A10-positive reactive astrocytes after spinal cord injury. Front Cell Neurosci. 17, 1276506 (2023). sourceCould not verify

  The Results (§3.3) discuss C3-positive reactive astrocytes, ApoE-related lipids, and inflammation after spinal cord injury; the Discussion (§4) likewise concerns spinal cord injury in mice. Searches for DAA, Alzheimer’s disease, APOE4, and iAstrocytes found no passage linking DAA pathway phenotypes to AD. (LLM, conf=0.76)

  Experimental — accuracy not yet measured
• cites [44] Wang, Y., Kang, X., Kang, X. & Yang, F. S100A6: molecular function and biomarker role. Biomark. Res. 11, 78 (2023). sourceCould not verify

  The 'Biomarker role of S100A6 in other diseases' section discusses reactive astrocytes in an ALS mouse model, not DAA phenotypes in Alzheimer’s disease. Searches for 'DAA,' 'Alzheimer,' and APOE4-related astrocyte inflammation found no passage in our copy addressing the attributed association. (LLM, conf=0.75)

  Experimental — accuracy not yet measured
• cites [45] Habib, N. et al. Disease-associated astrocytes in Alzheimer’s disease and aging. Nat. Neurosci. 23, 701–706 (2020). sourceSupported

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

APOE genetic variation alters lipid metabolism and fatty acid coupling in primary mouse neurons and astrocytes.

“One group found that APOE genetic variation alters lipid metabolism and fatty acid coupling in primary mouse neurons and astrocytes 31 .”

• cites [31] Qi, G. et al. ApoE4 Impairs Neuron-Astrocyte Coupling of Fatty Acid Metabolism. Cell Rep. 34, 108572 (2021). sourceSupported

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

The APOE4 allele is associated with altered synaptic density.

“Additionally, upregulation of synaptogenesis suggests a compensatory effect in APOE4 mice as previous work has found the APOE4 allele is associated with altered synaptic density [39][40][41] .”

• cites [39] He, K. et al. APOE epsilon4 is associated with decreased synaptic density in cognitively impaired participants. Alzheimers Dement 20, 3157–3166 (2024). sourceSupported
• cites [40] Zhao, J. et al. APOE4 exacerbates synapse loss and neurodegeneration in Alzheimer’s disease patient iPSC-derived cerebral organoids. Nat. Commun. 11, 5540 (2020). sourcePartly supported

  The Discussion reports that APOE4 exacerbated synaptic loss in AD patient-derived cerebral organoids at week 12. However, the Results measured synaptic protein levels rather than synaptic density and found no significant overall APOE4 effect on those levels. The source therefore supports an APOE4-related synaptic change in this model, but not the claim at its broader level of specificity. (LLM, conf=0.84)

  Experimental — accuracy not yet measured
• cites [41] Lin, Y. T. et al. APOE4 Causes Widespread Molecular and Cellular Alterations Associated with Alzheimer’s Disease Phenotypes in Human iPSC-Derived Brain Cell Types. Neuron 98, 1294 (2018). sourceNot checked — no full text of the source

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

Cfb has been previously linked to neurodegeneration.

“Interestingly, Cfb has been previously linked to neurodegeneration 38 .”

• cites [38] Mathys, H. et al. Temporal Tracking of Microglia Activation in Neurodegeneration at Single-Cell Resolution. Cell Rep. 21, 366–380 (2017). sourceSupported

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

APOE4 drives changes to lipid trafficking in primary mouse astrocytes.

“Another study found that APOE4 drives changes to lipid trafficking in primary mouse astrocytes 32 .”

• cites [32] Rawat, V. et al. ApoE4 Alters ABCA1 Membrane Trafficking in Astrocytes. J. Neurosci. 39, 9611–9622 (2019). sourceSupported

Classified as probably not a substantial claim.

Neurons have low expression of APOE under homeostatic conditions.

“Neurons have often been understudied in relation to the effects of APOE on metabolism due to their low expression of APOE under homeostatic conditions 36,37 .”

• cites [36] Zalocusky, K. A. et al. Neuronal ApoE upregulates MHC-I expression to drive selective neurodegeneration in Alzheimer’s disease. Nat. Neurosci. 24, 786–798 (2021). sourceSupported
• cites [37] Liu, L. et al. Apolipoprotein E expression is elevated by interleukin 1 and other interleukin 1-induced factors. J. Neuroinflammation 8, 175 (2011). sourceSupported

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

APOE genetic variation alters neuronal health and metabolism.

“In these limited studies, APOE genetic variation has been found to alter neuronal health and metabolism 30,31 .”

• cites [30] Budny V., et al. APOE4 Increases Energy Metabolism in APOE-Isogenic iPSC-Derived Neurons. Cells. 13, (2024). sourceSupported
• cites [31] Qi, G. et al. ApoE4 Impairs Neuron-Astrocyte Coupling of Fatty Acid Metabolism. Cell Rep. 34, 108572 (2021). sourceSupported

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

Observations of reduced APOE levels have been previously reported by other groups.

“Observations of reduced APOE levels have been previously reported by other groups 29,31 .”

• cites [29] Tcw, J. et al. Cholesterol and matrisome pathways dysregulated in astrocytes and microglia. Cell 185, 2213–33 e25 (2022). sourceSupported
• cites [31] Qi, G. et al. ApoE4 Impairs Neuron-Astrocyte Coupling of Fatty Acid Metabolism. Cell Rep. 34, 108572 (2021). sourceSupported

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

Microglia and astrocytes have APOE genotype-dependent immune and inflammatory metabolic changes.

“Additionally, microglia and astrocytes, which have been extensively studied in vivo, have APOE genotype dependent immune and inflammatory metabolic changes 22,25,26 .”

• cites [22] Lee, S. et al. APOE modulates microglial immunometabolism in response to age, amyloid pathology, and inflammatory challenge. Cell Rep. 42, 112196 (2023). sourceSupported
• cites [25] Liu, C. C. et al. Cell-autonomous effects of APOE4 in restricting microglial response in brain homeostasis and Alzheimer’s disease. Nat. Immunol. 24, 1854–1866 (2023). sourcePartly supported

  The Results sections on APOE isoform-dependent microglial signatures and human microglial lipid metabolism report genotype-dependent differences in immune responses and lipid accumulation. The Discussion mentions interactions with astrocytes but says that cell-type- and isoform-specific effects on glial function warrant further investigation; it does not establish the claimed changes in astrocytes. Page numbers were not available in our copy. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [26] Serrano-Pozo, A. et al. Effect of APOE alleles on the glial transcriptome in normal aging and Alzheimer’s disease. Nat. Aging 1, 919–931 (2021). sourcePartly supported

  The Results section reports APOE genotype-dependent proinflammatory gene expression in microglia and changes in astrocyte genes involved in lipid metabolism, with some astrocyte immune-response genes also identified. The Discussion characterizes the astrocyte association as more modest and chiefly involving lipid metabolism and extracellular-matrix genes, so the claim overstates a shared inflammatory pattern across both cell types. (LLM, conf=0.83)

  Experimental — accuracy not yet measured

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

Metabolite identities were predicted from exact mass, retention time compared with authentic standards, and tandem mass spectra fragmentation pattern compared with in-house standards and online databases.

“the identity predicted based on (i) the exact mass calculated from the mass per charge ratio; (ii) retention time compared to authentic standards, and (iii) the tandem mass spectra fragmentation pattern, compared to in-house standards, and online database 57,59,60 .”

• cites [57] Queathem, E. D. et al. Ketogenesis supports hepatic polyunsaturated fatty acid homeostasis via fatty acid elongation. Sci. Adv. 11, eads0535 (2025). sourceSupported
• cites [59] Huang, X. et al. X13CMS: global tracking of isotopic labels in untargeted metabolomics. Anal. Chem. 86, 1632–1639 (2014). sourcePartly supported

  The Experimental Section (LC/MS Analysis; page unavailable) reports m/z measurements and tandem-MS fragmentation analysis, and the Results and Discussion use fragmentation patterns to assess a labeled metabolite. The paper does not describe predicting metabolite identities by comparing retention times with authentic standards or fragmentation patterns with in-house standards and online databases; its retention-time alignment is used to group isotopologues. (LLM, conf=0.83)

  Experimental — accuracy not yet measured
• cites [60] Mahieu, N. G., Genenbacher, J. L. & Patti, G. J. A roadmap for the XCMS family of software solutions in metabolomics. Curr. Opin. Chem. Biol. 30, 87–93 (2016). sourcePartly supported

  The XCMS Online: Metabolomics on the Cloud section says uploaded MS and MS/MS data can be matched against the METLIN database using accurate mass and fragmentation patterns. It does not describe comparing retention times with authentic standards or fragmentation patterns with in-house standards, so those parts of the attributed method are unsupported. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

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

The decrease in PDH subunits and increase in PC in APOE4 iAstrocytes is consistent with a previous study.

“Surprisingly, we saw differences in these proteins including a decrease in subunits of PDH and an increase in PC in APOE4 iAstrocytes consistent with a previous study 35 .”

• cites [35] Lee, H. et al. ApoE4-dependent lysosomal cholesterol accumulation impairs mitochondrial homeostasis and oxidative phosphorylation in human astrocytes. Cell Rep. 42, 113183 (2023). sourcePartly supported

  The Results (page unavailable) report enhanced aerobic glycolysis and impaired mitochondrial respiration in ApoE4 astrocytes, broadly consistent with altered glucose metabolism. They do not report decreased PDH subunits or increased PC; searches for PDH and pyruvate carboxylase found no such comparison. (LLM, conf=0.78)

  Experimental — accuracy not yet measured

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

The upregulation of class I MHC mediated antigen processing and presentation pathways in APOE4 iNeurons is consistent with a previous study.

“We also found an upregulation of class I MHC mediated antigen processing and presentation pathways in APOE4 iNeurons which is consistent with a previous study 36 .”

• cites [36] Zalocusky, K. A. et al. Neuronal ApoE upregulates MHC-I expression to drive selective neurodegeneration in Alzheimer’s disease. Nat. Neurosci. 24, 786–798 (2021). sourceSupported

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

To enhance the capacity to identify chemical features, a standard mix of authentic standards for expected compounds was injected and pooled unlabeled samples were analyzed in triplicate by data-dependent analysis.

“To enhance our capacity to identify chemical features (i) a standard mix consisting of authentic standards for expected compounds was injected, and (ii) pooled samples of only unlabeled samples was analyzed in triplicate via data-dependent analysis mode (DDA) with IE omics scripts and R-Studio 58”

• cites [58] Koelmel, J. P. et al. Expanding Lipidome Coverage Using LC-MS/MS Data-Dependent Acquisition with Automated Exclusion List Generation. J. Am. Soc. Mass Spectrom. 28, 908–917 (2017). sourceSupported

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

The iPSCs were generated as previously described from the KOLF2.1 J parental cell line.

“These iPSCs were generated as previously described from the KOLF2.1 J parental cell line 54 .”

• cites [54] Pantazis, C. B. et al. A reference human induced pluripotent stem cell line for large-scale collaborative studies. Cell Stem Cell 29, 1685–702 e22 (2022). sourceSupported

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

The reliance on glucose for ATP production is consistent with past studies.

“This data tracks with our functional glycolytic assays and indicates a reliance on glucose for ATP production which is consistent with past studies 30,31,46 .”

• cites [30] Budny V., et al. APOE4 Increases Energy Metabolism in APOE-Isogenic iPSC-Derived Neurons. Cells. 13, (2024). sourcePartly supported

  The Results (§3.2; page unavailable) report glycolytic ATP production in the studied cells, supporting a role for glucose metabolism in ATP production. But the Discussion (§4; page unavailable) says the induced neurons primarily use oxidative phosphorylation under basal conditions; the paper does not establish a general reliance on glucose-driven glycolysis for ATP production. (LLM, conf=0.83)

  Experimental — accuracy not yet measured
• cites [31] Qi, G. et al. ApoE4 Impairs Neuron-Astrocyte Coupling of Fatty Acid Metabolism. Cell Rep. 34, 108572 (2021). sourceSupported
• cites [46] Fleeman, R. M., Kuhn, M. K., Chan, D. C. & Proctor, E. A. Apolipoprotein E epsilon4 modulates astrocyte neuronal support functions in the presence of amyloid-beta. J. Neurochem. 165, 536–549 (2023). sourcePartly supported

  The Results section ('Aβ treatment increases APOE4 astrocyte glycolytic ATP production'; page unavailable) reports increased glycolytic ATP production and mitochondrial glucose metabolism in APOE4 astrocytes exposed to Aβ. This supports glucose use for ATP production in that condition, but does not establish a general reliance on glucose or test whether ATP production depends on it. (LLM, conf=0.79)

  Experimental — accuracy not yet measured

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

The increased labeling of glutamic acid and aspartic acid is consistent with a past study in primary astrocytes.

“We found that both APOE4 iAstrocytes and iNeurons had increased labeling of glutamic acid and aspartic acid which is consistent with a past study in primary astrocytes 33 .”

• cites [33] Williams, H. C. et al. APOE alters glucose flux through central carbon pathways in astrocytes. Neurobiol. Dis. 136, 104742 (2020). sourcePartly supported

  The Results section ('APOE alters TCA cycle activity'; page unavailable) reports greater labeling of aspartate (m+4) and glutamate (m+4 and m+5) in E4 astrocytes. However, the Methods ('Tissue culture and in vitro [U-13C] glucose metabolite labeling'; page unavailable) identifies the cells as immortalized astrocytes derived from mice, not primary astrocytes. (LLM, conf=0.90)

  Experimental — accuracy not yet measured

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

Metabolite extraction and LC/MS analysis was performed as described previously.

“Metabolite extraction and LC/MS analysis was performed as described previously [55][56][57] .”

• cites [55] Puchalska, P. et al. Isotope Tracing Untargeted Metabolomics Reveals Macrophage Polarization-State-Specific Metabolic Coordination across Intracellular Compartments. iScience 9, 298–313 (2018). sourcePartly supported

  The Introduction describes LC–high-resolution MS-based untargeted metabolomics, supporting the citation for LC/MS analysis. The Methods section directs readers to a supplemental file, but our copy does not describe metabolite extraction or establish that the citing paper followed the same procedures. (LLM, conf=0.78)

  Experimental — accuracy not yet measured
• cites [56] Puchalska, P. et al. Hepatocyte-Macrophage Acetoacetate Shuttle Protects against Tissue Fibrosis. Cell Metab. 29, 383–98 e7 (2019). sourceSupported
• cites [57] Queathem, E. D. et al. Ketogenesis supports hepatic polyunsaturated fatty acid homeostasis via fatty acid elongation. Sci. Adv. 11, eads0535 (2025). sourceSupported

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

UDP-GlcNAc participates in numerous cellular functions including nutrient sensing and glycosylation.

“UDP-GlcNAc participates in numerous cellular functions including nutrient sensing and glycosylation 47 .”

• cites [47] Mannino, M. P. & Hart, G. W. The Beginner’s Guide to O-GlcNAc: From Nutrient Sensitive Pathway Regulation to Its Impact on the Immune System. Front Immunol. 13, 828648 (2022). sourceSupported

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

Quality control and normalization of the proteomics data was carried out with proteiNorm.

“Quality control (QC) and normalization was carried out with proteiNorm 61 .”

• cites [61] Graw, S. et al. proteiNorm - A User-Friendly Tool for Normalization and Analysis of TMT and Label-Free Protein Quantification. ACS Omega 5, 25625–25633 (2020). sourceSupported

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

These findings of decreased mitochondrial function and a switch to glycolysis are consistent with previous studies.

“These findings of decreased mitochondrial function and a switch to glycolysis is consistent with previous studies 33,35 .”

• cites [33] Williams, H. C. et al. APOE alters glucose flux through central carbon pathways in astrocytes. Neurobiol. Dis. 136, 104742 (2020). sourcePartly supported

  The Discussion reports increased glucose flux into oxygen-independent glycolysis in E4 astrocytes, and the Conclusions report increased glucose flux into lactate. Those passages support the glycolysis part, but they do not establish decreased mitochondrial function; the paper instead describes a different pattern of TCA-cycle activity in E4 astrocytes. No page numbers are available in our copy. (LLM, conf=0.83)

  Experimental — accuracy not yet measured
• cites [35] Lee, H. et al. ApoE4-dependent lysosomal cholesterol accumulation impairs mitochondrial homeostasis and oxidative phosphorylation in human astrocytes. Cell Rep. 42, 113183 (2023). sourceSupported

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

APOE TR mice were previously generated by removing and replacing the endogenous mouse Apoe gene with the human APOE gene.

“These APOE TR mice were previously generated by removing and replacing the endogenous mouse Apoe gene with the human APOE gene 51,52 .”

• cites [51] Sullivan, P. M. et al. Targeted replacement of the mouse apolipoprotein E gene with the common human APOE3 allele enhances diet-induced hypercholesterolemia and atherosclerosis. J. Biol. Chem. 272, 17972–17980 (1997). sourceNot checked — no full text of the source
• cites [52] Knouff, C. et al. Apo E structure determines VLDL clearance and atherosclerosis risk in mice. J. Clin. Invest 103, 1579–1586 (1999). sourceSupported

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

Decreased O-GlcNAcylation is associated with AD diagnosis and neurodegeneration.

“This is particularly interesting since decreased O-GlcNAcylation is associated with AD diagnosis and neurodegeneration [48][49][50] .”

• cites [48] Park J., et al. O-GlcNAcylation ameliorates the pathological manifestations of Alzheimer’s disease by inhibiting necroptosis. Sci. Adv.7, (2021). sourceSupported
• cites [49] Chen, L., Jiang, H., Licinio, J. & Wu, H. Brain O-GlcNAcylation: Bridging physiological functions, disease mechanisms, and therapeutic applications. Mol. Psychiatry 30, 2754–2772 (2025). sourceSupported
• cites [50] Wang, A. C., Jensen, E. H., Rexach, J. E., Vinters, H. V. & Hsieh-Wilson, L. C. Loss of O-GlcNAc glycosylation in forebrain excitatory neurons induces neurodegeneration. Proc. Natl. Acad. Sci. USA. 113, 15120–15125 (2016). sourcePartly supported

  The paper reports that loss of OGT in adult mouse forebrain neurons causes progressive neurodegeneration. It also reports lower OGT protein expression in cortical neurons from patients with severe AD (Fig. 6), but it does not establish that O-GlcNAcylation itself is decreased in those patients; the human-brain results section notes prior reports of both increases and decreases in O-GlcNAcylated proteins. Page numbers are unavailable in our copy. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

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

MitoCarta 3.0 was used to identify mitochondrial-specific proteins in the isolated brain mitochondria samples.

“MitoCarta 3.0 was used to further identify mitochondrial specific proteins in the isolated brain mitochondria samples 62 .”

• cites [62] Rath, S. et al. MitoCarta3.0: an updated mitochondrial proteome now with sub-organelle localization and pathway annotations. Nucleic Acids Res. 49, D1541–D1547 (2021). sourceSupported

May combine two claims.

Interferon signaling was significantly upregulated in iNeurons with APOE4 in addition to other antiviral inflammatory proteins.

“Interferon signaling was significantly upregulated in iNeurons with APOE4 in addition to other antiviral inflammatory proteins.”

May combine two claims.

Mitochondrial membrane potential, mitochondrial superoxide production, H2O2, intracellular Ca2+, and mitochondrial Ca2+ were all increased in both Pair A and B derived APOE4 iAstrocytes.

“Mitochondrial membrane potential, mitochondrial superoxide production, hydrogen peroxide (H 2 O 2 ), intracellular calcium (Ca 2+ ), and mitochondrial Ca 2+ were all increased in both Pair A and B derived APOE4 iAstrocytes (Fig. 4G, H).”

May combine two claims.

APOE4 females had reduced expression of proteins involved in neurite growth/axonal guidance, protein quality control, Jak/Stat and tyrosine receptor signaling, glutamate receptor signaling, coenzyme Q synthesis, and centriole duplication.

“There was reduced expression of proteins involved in neurite growth/axonal guidance (Myo9a), protein quality control (Selenof), Jak/Stat and tyrosine receptor signaling (Jak1), glutamate receptor signaling (Grik5), coenzyme Q synthesis (Coq3), and centriole duplication (Mdm1) (Fig. 1C).”

May combine two claims.

APOE4 iNeurons showed a decrease in protein expression of PC and no difference in PDC subunits.

“APOE4 iNeurons showed a decrease in protein expression of PC and no difference in PDC subunits (Fig. 8G, H).”

May combine two claims.

APOE4 iNeurons had very modestly decreased unlabeled aspartic acid and increased labeling with M+4 and M+5 isotopologues for glutamic acid but decreased labeling of M+1 isotopologue.

“APOE4 iNeurons had very modestly decreased unlabeled aspartic acid and increased labeling with M + 4 and M + 5 isotopologues for glutamic acid but decreased labeling of the M + 1 isotopologue (Fig. 7G, H).”

May combine two claims.

M+3 isotopologue of malic acid was less enriched in APOE4 iAstrocytes than APOE3, while M+4 showed greater enrichment.

“The M + 3 isotopologue of malic acid was less enriched in APOE4 iAstrocytes than in APOE3 iAstrocytes, while the M + 4 showed greater enrichment (Fig. 7E).”

May combine two claims.

Mitochondrial membrane potential, H2O2, and total cellular calcium were increased in Pair B APOE4 iNeurons but not Pair A.

“Mitochondrial membrane potential, H 2 O 2 , and total cellular calcium were increased in Pair B APOE4 iNeurons but not Pair A (Fig. 5G, H).”

May combine two claims.

Male APOE4 mice showed increased expression of proteins involved in fatty acid metabolism, ER/Golgi maintenance, complement and cell death/inflammation, and neuronal development/inhibitory presynaptic differentiation.

“When compared to their APOE3 counterparts, male APOE4 mice showed divergence from the APOE4 versus APOE3 comparison in females, with increased expression of proteins involved in fatty acid metabolism (Acsl5), endoplasmic reticulum (ER)/Golgi maintenance (Yif1a), complement and cell death/inflammation (Cfb, Gsdma), and neuronal development/inhibitory presynaptic differentiation (lgsf21, Mycbp2).”

May combine two claims.

O-GlcNAc levels showed a decreased trend in APOE4 iAstrocytes and were significantly reduced in APOE4 iNeurons.

“Although proteins in this pathway were increased, O-GlcNAc levels showed a decreased trend in APOE4 iAstrocytes and were significantly reduced in APOE4 iNeurons (Fig. 9L).”

May combine two claims.

Proteins down regulated in APOE4 iNeurons included axon guidance (PLPPR3, SPON1), actin/cytoskeleton organization and cell barrier (MYH14, EPPK1, CLDN6), lipid metabolism (LPL), serotonin synthesis (TPH2), and insulin like growth factor signaling (IGFBP7).

“Proteins that were down regulated in APOE4 iNeurons included those associated with axon guidance (PLPPR3, SPON1), actin/cytoskeleton organization and cell barrier (MYH14, EPPK1, CLDN6), lipid metabolism (LPL), serotonin synthesis (TPH2), and insulin like growth factor signaling (IGFBP7) (Fig. 3F).”

May combine two claims.

Mitochondrial superoxide was increased in Pair A APOE4 iNeurons with an increased trend in Pair B.

“Mitochondrial superoxide was increased in Pair A APOE4 iNeurons with an increased trend in Pair B (Fig. 5G, H).”

May combine two claims.

We see a downregulation of pathways involved in complex I biogenesis, respiratory chain/oxidative phosphorylation, and mitochondrial translation.

“These changes in protein expression are interesting since we see a downregulation of pathways involved in complex I biogenesis, respiratory chain/oxidative phosphorylation, and mitochondrial translation.”

May combine two claims.

Proteomic analysis of isolated mitochondria from brain revealed down regulation of lipid metabolism pathway components (Phyh in males, Lypla1 in females) with upregulation of certain electron transport chain components (Nubpl in females, Cox7a1 in males).

“Proteomic analysis of isolated mitochondria from brain further revealed a down regulation of lipid metabolism pathway components (Phyh in males, Lypla1 in females) with an upregulation of certain electron transport chain components (Nubpl in females, Cox7a1 in males).”

May combine two claims.

Analysis of TCA cycle proteins revealed an overall decrease in APOE4 iAstrocytes, while APOE4 iNeurons exhibited increases.

“Analysis of TCA cycle proteins revealed an overall decrease in APOE4 iAstrocytes, while APOE4 iNeurons exhibited increases (Fig. 8A-C).”

May combine two claims.

Down regulated pathways included glutamate degradation, apoptosis, CLEAR signaling, sirtuin signaling, and mitochondrial dysfunction pathways in APOE4 male mice compared to APOE3.

“Down regulated pathways included glutamate degradation, apoptosis, CLEAR signaling, sirtuin signaling, and mitochondrial dysfunction pathways in APOE4 male mice when compared to APOE3 (Fig. 1F).”

May combine two claims.

IPA showed upregulation of sirtuin signaling, granzyme A signaling, mitochondrial dysfunction, protein sorting, RHOA signaling, COPI anterograde transport, Parkinson's signaling, and nucleotide biosynthesis pathways in APOE4 iAstrocytes.

“IPA showed an upregulation of sirtuin signaling, granzyme A signaling, mitochondrial dysfunction, protein sorting, RHOA signaling, COPI anterograde transport, Parkinson's signaling, and nucleotide biosynthesis pathways in APOE4 iAstrocytes (Fig. 3G).”

May combine two claims.

Using IPA we found mitochondrial dysfunction pathway upregulated with cristae formation, complex I biogenesis, and respiratory chain/oxidative phosphorylation being downregulated in APOE4 derived iAstrocytes.

“Using IPA we found that the mitochondrial dysfunction pathway was upregulated with cristae formation, complex I biogenesis, and respiratory chain/oxidative phosphorylation being downregulated in APOE4 derived iAstrocytes.”

May combine two claims.

iNeurons had decreased glucose and glutamine fuel dependency with increased capacity to use glucose and decreased capacity to use fatty acids as fuel sources.

“iNeurons had decreased glucose and glutamine fuel dependency with increased capacity to use glucose and decreased capacity to use fatty acids as fuel sources (Fig. 6G, H).”

May combine two claims.

Proteins down regulated in APOE4 iAstrocytes included axon guidance/VEGF receptor ligand (NRP2), protein trafficking (SORT1), synaptogenesis (NCAM1), chaperone/lipid transport (CLU), lipid droplet formation (BSCL2), and ER membrane trafficking and stress (RTN1).

“Proteins that were down regulated in APOE4 iAstrocytes included those associated with axon guidance/VEGF receptor ligand (NRP2), protein trafficking (SORT1), synaptogenesis (NCAM1), chaperone/lipid transport (CLU), lipid droplet formation (BSCL2), and ER membrane trafficking and stress (RTN1) (Fig. 3E).”

May combine two claims.

Unlabeled succinate was significantly increased in APOE4 iAstrocytes with reduced labeling for M+2 isotopologue.

“Unlabeled succinate was significantly increased in APOE4 iAstrocytes with reduced labeling for the M + 2 isotopologue (Fig. 7D).”

May combine two claims.

Unlabeled succinate was increased in APOE4 iNeurons with diminished labeling of M+1, M+2, M+3, and M+4 isotopologues.

“Unlabeled succinate was increased in APOE4 iNeurons with diminished labeling of the M + 1, M + 2, M + 3, and M + 4 isotopologues (Fig. 7I).”

May combine two claims.

Compared to female APOE3 mice, APOE4 females had increased expression of proteins involved in mitochondrial function and bioenergetics, glucose metabolism, ubiquitination/inflammation, peptidase activity, and clathrin recruitment/receptor sorting.

“Compared to female APOE3 mice, APOE4 females had increased expression of proteins involved in mitochondrial function and bioenergetics (mt-Co3, Sdhd, Adcy3), glucose metabolism (Slc2a1/Glut1), ubiquitination/inflammation (Otulin), peptidase activity (Serpina1e), and clathrin recruitment/receptor sorting (Ap1s2).”

May combine two claims.

Carbohydrate driven respiration showed a main effect for genotype and an interaction between sex and genotype for state 3 respiration.

“Carbohydrate driven respiration showed a main effect for genotype and an interaction between sex and genotype for state 3 respiration.”

May combine two claims.

MRPL47 and MRPS23 were down regulated in female APOE4 mice while MRSP2 was upregulated in male APOE4 mice.

“Specific mitochondrial ribosomal proteins including MRPL47 and MRPS23 were down regulated in female APOE4 mice while MRSP2 was upregulated in male APOE4 mice (Fig. 2J).”

May combine two claims.

APOE protein expression in APOE4 iAstrocytes was decreased compared to APOE3 cells, an effect not observed in iNeurons.

“APOE protein expression in APOE4 iAstrocytes was decreased when compared to APOE3 cells, an effect not observed in iNeurons, which showed no difference (Fig. 3C, D).”

May combine two claims.

APOE4 iAstrocytes showed upregulation of 2344 proteins (33.40%) and down regulation of 2463 proteins (35.09%).

“Surprisingly, APOE4 iAstrocytes showed an upregulation of 2344 proteins (33.40%) and a down regulation of 2463 proteins (35.09%) (Fig. 3E).”

May combine two claims.

Male APOE4 mice had reduced expression of proteins involved in lipid metabolism, endothelial cell function/VEGF signaling, iron/sulfur assembly, and synaptophysin like/obesity linked proteins.

“There was reduced expression of proteins involved in lipid metabolism (Fabp5, Apod), endothelial cell function/VEGF signaling (Edf1), iron/sulfur assembly (Nubpl), and synaptophysin like/obesity linked proteins (Sypl2) (Fig. 1E).”

May combine two claims.

Genotype-dependent effects were less frequently observed, and lower in magnitude, in iNeurons than in iAstrocytes.

“While overall labeling was increased in iNeurons than in iAstrocytes, genotype-dependent effects were less frequently observed, and lower in magnitude, in iNeurons than in iAstrocytes.”

May combine two claims.

APOE4 iAstrocytes showed reduced complex I, II, and IV flux/driven respiration in both isogenic pairs, with disparate results for complex III (reduced in Pair A).

“APOE4 iAstrocytes also showed reduced complex I, II, and IV flux/ driven respiration in both isogenic pairs, with disparate results for complex III (reduced in Pair A) (Fig. 4E, F).”

May combine two claims.

APOE4 iAstrocytes had elevated M+4 malate and reduced protein levels of PDC subunits.

“In contrast, while APOE4 iAstrocytes had elevated M + 4 malate, protein levels of pyruvate dehydrogenase complex (PDC) subunits were reduced, indicating impaired glucose-derived carbon entry into the TCA cycle (Fig. 7E and Fig. 8F).”

May combine two claims.

Oxidative phosphorylation proteins were largely down regulated in APOE4 iAstrocytes while being slightly upregulated in APOE4 iNeurons.

“Interestingly, oxidative phosphorylation proteins were largely down regulated in APOE4 iAstrocytes while being slightly upregulated in APOE4 iNeurons (Fig. 3I).”

May combine two claims.

Selected proteins upregulated in APOE4 iNeurons included inflammation (IFI35, IFITMI), extracellular matrix (DCN), calcium signaling (ITPKB), neuronal signaling/metalloenzyme (CA2), and axon regeneration/neurotrophic factors (DLK1, FGF1).

“Selected proteins that were upregulated in APOE4 iNeurons included those associated with inflammation (IFI35, IFITMI), extracellular matrix (DCN), calcium signaling (ITPKB), neuronal signaling/metalloenzyme (CA2) and axon regeneration/neurotrophic factors (DLK1, FGF1) (Fig. 3F).”

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

At a pathway level, sex differences seemed less pronounced in APOE4 mice.

“One intriguing observation was that at a pathway level, sex differences seemed to be less pronounced in APOE4 mice (Supplemental Fig. 1D, E).”

May combine two claims.

APOE4 iAstrocytes had increased labeling of M+1, M+2, M+4, and M+5 isotopologues for glutamic acid but decreased labeling of M+3 isotopologue.

“Additionally, there was increased labeling of the M + 1, M + 2, M + 4, and M + 5 isotopologues for glutamic acid but decreased labeling of the M + 3 isotopologue in APOE4 iAstrocytes (Fig. 7C).”

May combine two claims.

Proteins involved in mitochondrial DNA maintenance/GDP formation (Guk1), α-oxidation/lipid metabolism (Phys), ubiquitin ligase (Mul1), and viral RNA degradation/AMP metabolism (Nudt2) were down regulated in male APOE4 mice compared to APOE3.

“Proteins involved in mitochondrial DNA maintenance/GDP formation (Guk1), α-oxidation/lipid metabolism (Phys), ubiquitin ligase (Mul1), and viral RNA degradation/AMP metabolism (Nudt2) were down regulated in male APOE4 mice when compared to APOE3 males (Fig. 2G).”

May combine two claims.

Carbohydrate driven respiration showed an interaction between sex and genotype for state 3 G with increased respiration in male APOE3 mice and a trend for decreased state 3S respiration in APOE4 mice.

“Carbohydrate driven respiration showed an interaction between sex and genotype for state 3 G with increased respiration in male APOE3 mice and a trend for decreased state 3S respiration in APOE4 mice (Fig. 2C, D).”

May combine two claims.

Glucose and fatty acid fuel source dependency and capacity were both increased in APOE4 iAstrocytes.

“Glucose and fatty acid fuel source dependency and capacity were both increased in APOE4 iAstrocytes (Fig. 6E, F).”

May combine two claims.

APOE4 derived iNeurons increased inflammatory protein expression and calcium signaling with decreased expression of proteins involved in lipid metabolism (specifically CLU and SORT1).

“APOE4 derived iNeurons also increased inflammatory protein expression and calcium signaling with decreased expression of proteins involved in lipid metabolism (specifically CLU and SORT1).”

May combine two claims.

Male APOE4 mice had increased expression of metalloendopeptidase (Nrdc), cytochrome oxidase (Cox7a1), mitochondrial DNA integrity/mitophagy inhibition (Fbxl14), and mitochondrial ribosome (Mrps2) proteins.

“Male APOE4 mice had increased expression of metalloendopeptidase (Nrdc), cytochrome oxidase (Cox7a1), mitochondrial DNA integrity/mitophagy inhibition (Fbxl14), and mitochondrial ribosome (Mrps2) proteins (Fig. 2G).”

May combine two claims.

Male APOE4 mice had reduced activation of mitochondrial dysfunction pathway with increased activation of pathways for cristae formation.

“Male APOE4 mice also had reduced activation of the mitochondrial dysfunction pathway with increased activation of pathways for cristae formation, suggesting an effort to compensate for metabolic stress or reduced mitochondrial mass.”

May combine two claims.

Mitochondrial ribosome (Mrpl55), RNA methylation (Mett15), lysophospholipase activity (Lypla1), and iron homeostasis (Fth1) proteins were down regulated in female APOE4 mice.

“Mitochondrial ribosome (Mrpl55), RNA methylation (Mett15), lysophospholipase activity (Lypla1), and iron homeostasis (Fth1) proteins were down regulated in female APOE4 mice (Fig. 2F).”

May combine two claims.

Glycolytic flux was increased in Pair A APOE4 iAstrocytes, including glycolytic capacity, glycolytic reserve, and a trend for increased non-glycolytic extracellular acidification rate.

“Glycolytic flux was increased in Pair A APOE4 iAstrocytes, including glycolytic capacity, glycolytic reserve, and a trend for increased non-glycolytic extracellular acidification rate (Fig. 6A, B).”

May combine two claims.

Despite both cell types showing profound metabolic stress and mitochondrial dysfunction, oxidative phosphorylation associated proteins were downregulated in APOE4 iAstrocytes and upregulated in APOE4 iNeurons.

“Despite both cell types showing profound metabolic stress and mitochondrial dysfunction, oxidative phosphorylation associated proteins were downregulated in APOE4 iAstrocytes and upregulated in APOE4 iNeurons.”

Classified as probably not a substantial claim.

This study did not report outcomes in iNeurons where we observed an increase in overall expression of TCA cycle proteins.

“This study did not report outcomes in iNeurons where we observed an increase in overall expression of TCA cycle proteins.”

May combine two claims.

Functional mitochondria in iAstrocytes and iNeurons revealed reduced mitochondrial respiration and electron transport chain function in APOE4 derived cells with increased indices of oxidative stress and mitochondrial calcium.

“Functional mitochondria in iAstrocytes and iNeurons revealed reduced mitochondrial respiration and electron transport chain function in APOE4 derived cells with increased indices of oxidative stress and mitochondrial calcium.”

May combine two claims.

Reductions in mitochondrial respiration were coupled with increased glycolytic flux and increased dependency/capacity on glucose fuel use in iAstrocytes with APOE4.

“Reductions in mitochondrial respiration were coupled with increased glycolytic flux and increased dependency/capacity on glucose fuel use in iAstrocytes with APOE4.”

May combine two claims.

We saw differences in TCA cycle and pyruvate metabolism proteins including a decrease in subunits of PDH and an increase in PC in APOE4 iAstrocytes.

“Surprisingly, we saw differences in these proteins including a decrease in subunits of PDH and an increase in PC in APOE4 iAstrocytes consistent with a previous study 35 .”

May combine two claims.

We observed changes in whole brain metabolism and mitochondrial function, including cell type-specific alterations in the absence of amyloid or tau pathology.

“We observed changes in whole brain metabolism and mitochondrial function, including cell type-specific alterations in the absence of amyloid or tau pathology.”

May combine two claims.

Data in APOE4 iNeurons showed a reduction in glucose dependency and an increase in capacity which was paired with reduced GLUT3 protein expression.

“However, data in APOE4 iNeurons showed a reduction in glucose dependency and an increase in capacity which was paired with reduced GLUT3 protein expression.”

May combine two claims.

While APOE4 iAstrocytes and iNeurons displayed activation of inflammatory pathways and a metabolic shift towards glycolysis, changes in mitochondrial dysfunction were disparate between cell types.

“Additionally, while APOE4 iAstrocytes and iNeurons displayed activation of inflammatory pathways and a metabolic shift towards glycolysis, changes in mitochondrial dysfunction were disparate between cell types.”

May combine two claims.

Cholesterol biosynthesis and insulin like growth factor signaling were all downregulated in APOE4 iNeurons.

“Cholesterol biosynthesis, insulin like growth factor signaling were all downregulated in APOE4 iNeurons.”

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based). May combine two claims.

The significance of reduced APOE4 expression includes dysfunctional lipid trafficking and/or reduced lipid metabolism.

“The significance of reduced APOE4 expression includes dysfunctional lipid trafficking and/or reduced lipid metabolism.”

May combine two claims.

APOE genotype caused extensive changes to metabolism consistently across in vivo and in vitro models, including sex-specific changes in APOE TR mice.

“We found that APOE genotype caused extensive changes to metabolism consistently across in vivo and in vitro models, including important sex specific changes in APOE TR mice.”

May combine two claims.

These data revealed reduced APOE protein expression in APOE4 mouse brain and iAstrocytes, but not iNeurons.

“These data revealed reduced APOE protein expression in APOE4 mouse brain and iAstrocytes, but not iNeurons.”

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

Animal work and protocols were conducted under approval from the Institutional Animal Care and Use Committee at the University of Kansas Medical Center.

“Animal work and protocols were conducted under approval from the Institutional Animal Care and Use Committee at the University of Kansas Medical Center Assurance Number: A3237-01.”

May combine two claims.

Collectively, our findings suggest that APOE4 suppresses mitochondrial TCA cycle activity in iAstrocytes and increases it in iNeurons.

“Collectively, our findings suggest that APOE4 suppresses mitochondrial TCA cycle activity in iAstrocytes and increases it in iNeurons.”

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

Raw metabolomics data was processed by removing background ions, aligning retention times of detected signals across all samples, and grouping chemical features.

“Raw data was uploaded, background ions removed, retention time for detected signals aligned across all samples, chemical features grouped”

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

The stable isotope enrichment of each metabolic pool was determined by Compound Discoverer after correction for natural abundance.

“The stable isotope enrichment of each metabolic pool was determined by Compound Discoverer after the correction for natural abundance.”

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

Outliers were identified and removed from datasets using Grubbs' test with a significant alpha of 0.05.

“Outliers were identified and removed from datasets using Grubbs' test with a significant alpha of 0.05.”

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

Another limitation is the use of humanized mouse models, as expression of a human protein in a mouse often leads to functional differences that are not physiological.

“Another limitation is the use of humanized mouse models, as expression of a human protein in a mouse often leads to functional differences that are not physiological.”

• AssumptionRests on an assumption: Expression of a human protein in a mouse often leads to functional differences that are not physiological.

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

Other studies have examined the effects of APOE in mouse models that produce amyloid and tau.

“The mouse models we examined do not produce hallmark AD pathology, but other studies have examined the effects of APOE in mouse models that produce amyloid and tau.”

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

This study has a limitation of lack of co-culture experiments between neurons and astrocytes.

“The first limitation is the lack of co-culture experiments between neurons and astrocytes.”

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

Respiration values were normalized to total protein loaded into the chamber and across days of data acquisition.

“Respiration values were normalized to total protein loaded into chamber and across days of data acquisition.”

• Also stated in: § Statistical analysis · plain

  “Respiration data was normalized across each day of data acquisition.”

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

Protein loading was normalized by BCA assay and an equal amount of protein was resolved on the gels.

“A BCA assay (ThermoFisher) was performed to normalize protein loading, and an equal amount of protein was resolved on 4-15% Criterion TGX gels (Bio-Rad).”

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

For the isotope tracing untargeted metabolomics pipeline, blanks and pooled quality control samples were injected throughout the batch run.

“For ITUM pipeline, both blanks and pooled quality control (QC) samples were injected throughout the batch run.”

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

Fluorescence intensity for each dye was normalized to Hoechst.

“Fluorescence intensity for each dye was read with a Tecan Infinite 200 PRO® and normalized to Hoechst.”