Small Molecule Structure Correctors Abolish Detrimental Effects of Apolipoprotein E4 in Cultured Neurons
Open accessAbstract
Background: Apolipoprotein E4 (apoE4), the major gene involved in Alzheimer disease, has a unique structure, intramolecular domain interaction, that is associated with neuropathology.Results: Potent small molecule structure correctors block apoE4 domain interaction and reverse apoE4 detrimental effects in cultured neurons.Conclusion: Structure correctors negate the detrimental effects of apoE4 in neurons.Significance: ApoE4 structure correctors could represent a therapeutic approach for treating apoE4-associated neuropathology.Apolipoprotein E4 (apoE4), the major genetic risk factor for late onset Alzheimer disease, assumes a pathological conformation, intramolecular domain interaction.ApoE4 domain interaction mediates the detrimental effects of apoE4, including decreased mitochondrial cytochrome c oxidase subunit 1 levels, reduced mitochondrial motility, and reduced neurite outgrowth in vitro.Mutant apoE4 (apoE4-R61T) lacks domain interaction, behaves like apoE3, and does not cause detrimental effects.To identify small molecules that inhibit domain interaction (i.e.structure correctors) and reverse the apoE4 detrimental effects, we established a high throughput cell-based FRET primary assay that determines apoE4 domain interaction and secondary celland function-based assays.Screening a ChemBridge library with the FRET assay identified CB9032258 (a phthalazinone derivative), which inhibits domain interaction in neuronal cells.In secondary functional assays, CB9032258 restored mitochondrial cytochrome c oxidase subunit 1 levels and rescued impairments of mitochondrial motility and neurite outgrowth in apoE4-expressing neuronal cells.These benefits were apoE4specific and dose-dependent.Modifying CB9032258 yielded well defined structure-activity relationships and more active compounds with enhanced potencies in the FRET assay (IC 50 of 23 and 116 nM, respectively).These compounds efficiently restored functional activities of apoE4-expressing cells in secondary assays.An EPR binding assay showed that the apoE4 structure correction resulted from direct interaction of a phthalazinone.With these data, a six-feature pharmacophore model was constructed for future drug design.Our results serve as a proof of concept that pharmacological intervention with apoE4 structure correctors negates apoE4 detrimental effects in neuronal cells and could be further developed as an Alzheimer disease therapeutic.
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Small-molecule structure correctors have been shown to restore ApoE3-like behavior in iPSC-derived neurons.
“Such correctors have already been shown to restore ApoE3-like behavior in iPSC-derived neurons (37,48)”
From APOE Isoform-Dependent Self-Association Measured by a Split-Luciferase Complementation Assay: Differential Effects of Disease-Risk and Protective Variants (2026) · cites this paper as [37]
The Discussion reports that structure correctors restored ApoE4-related functions toward ApoE3-like behavior in cultured neurons. The Experimental Procedures identify the cells as Neuro-2a and PC12 cells and primary neurons from transgenic mice, not iPSC-derived neurons; that population-specific part of the claim is unsupported. (LLM, conf=0.90)
Experimental — accuracy not yet measuredThe aberrant proteolytic processing of apoE4 and consequent neurotoxicity can be blocked by treating apoE4-expressing cells with small-molecule structure correctors, which convert apoE4 to an apoE3-like conformation and prevent apoE4 fragment generation.
“The aberrant proteolytic processing of apoE4 and consequent neurotoxicity can be blocked by treating apoE4-expressing cells with small-molecule “structure correctors,” which convert apoE4 to an apoE3-like conformation and prevent apoE4 fragment generation [12, 13, 18, 19].”
From Neuronal Apolipoprotein E4 Expression Results in Proteome-Wide Alterations and Compromises Bioenergetic Capacity by Disrupting Mitochondrial Function (2019) · cites this paper as [19]
Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.
Experimental — accuracy not yet measuredApoE fragments engage in abnormal interactions in the cytosol that lead to neurodegeneration.
“where they engage in abnormal interactions that lead to neurodegeneration [17–19]”
From Neuronal Apolipoprotein E4 Expression Results in Proteome-Wide Alterations and Compromises Bioenergetic Capacity by Disrupting Mitochondrial Function (2019) · cites this paper as [19]
The Discussion (page unavailable) identifies neurotoxic apoE4 fragment formation among apoE4’s neuropathological features, and describes an abnormal interaction between domains of apoE4. It does not say that the fragments engage in abnormal interactions in the cytosol or that those interactions lead to neurodegeneration. (LLM, conf=0.85)
Experimental — accuracy not yet measuredMultiple small molecules have been designed to correct the conformation of APOE4 so that it resembles that of APOE3 or APOE2.
“Multiple small molecules have been designed to ‘correct’ the conformation of APOE4 so that it resembles that of APOE3 or APOE2 (refs. 213–217)”
From Multifaceted roles of APOE in Alzheimer disease (2024) · cites this paper as [214]
In neurons, apoE4 is associated with impaired mitochondrial respiration.
“In neurons, apoE4 is associated with impaired mitochondrial respiration”
From Neuronal Apolipoprotein E4 Expression Results in Proteome-Wide Alterations and Compromises Bioenergetic Capacity by Disrupting Mitochondrial Function (2019) · cites this paper as [19]
In neurons, apoE4 is associated with decreased neurite outgrowth.
“decreased neurite outgrowth”
From Neuronal Apolipoprotein E4 Expression Results in Proteome-Wide Alterations and Compromises Bioenergetic Capacity by Disrupting Mitochondrial Function (2019) · cites this paper as [19]
In neurons, apoE4 is associated with neurotoxicity.
“and neurotoxicity [10, 17–19, 34, 35]”
From Neuronal Apolipoprotein E4 Expression Results in Proteome-Wide Alterations and Compromises Bioenergetic Capacity by Disrupting Mitochondrial Function (2019) · cites this paper as [19]
ApoE fragments escape the secretory pathway into the cytosol.
“The resulting apoE fragments escape the secretory pathway into the cytosol”
From Neuronal Apolipoprotein E4 Expression Results in Proteome-Wide Alterations and Compromises Bioenergetic Capacity by Disrupting Mitochondrial Function (2019) · cites this paper as [19]
The Discussion (page not available) mentions neuron-specific proteolysis and neurotoxic apoE4 fragment formation. Neither that passage nor passages retrieved for “cytosol” and “secretory pathway” say that apoE fragments leave the secretory pathway or enter the cytosol. (LLM, conf=0.74)
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- Authors
- Chen, H.-K., Liu, Z., Meyer-Franke, A., Brodbeck, J., Miranda, R. D., McGuire, J. G., Pleiss, M. A., Ji, Z.-S., Balestra, M. E., Walker, D. W., Xu, Q., Jeong, D.-E., Budamagunta, M. S., Voss, J. C., Freedman, S. B., Weisgraber, K. H., Huang, Y., & Mahley, R. W.
- Affiliations
- Gladstone Institutes (US)
- University of California, Davis (US)
- Corresponding author
- Mahley, Robert W.
- ORCID
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2012-02
- Volume
- 287
- Issue
- 8
- Pages
- 5253-5266
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC3285306
- OpenAlex
- W2014055987
- Cited by
- 163
- References
- 89
- Citations by year
- 2026: 15 · 2025: 13 · 2024: 13 · 2023: 8 · 2022: 10 · 2021: 12 · 2020: 14 · 2019: 9 · 2018: 11 · 2017: 5 · 2016: 15 · 2015: 5 · 2014: 11 · 2013: 11 · 2012: 10
- FWCI
- 6.98
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (AG‐022074)
- National Institute on Aging (R01AG029246, P01AG022074)
- Topics
- Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
- Health Sciences › Medicine › Psychiatry and Mental health › Dementia and Cognitive Impairment Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
- Keywords
- APOE ε4, Alzheimer's disease, small molecule inhibitors, phthalazinone derivatives, mitochondrial cytochrome c oxidase subunit 1, mitochondrial motility, neurite outgrowth, structure-activity relationship, pharmacophore model, FRET assay, cultured neurons
- MeSH terms
- Cell Line, Drug Evaluation, Preclinical, Humans, Models, Molecular, Neurons, Phthalazines, Structure-Activity Relationship, Reproducibility of Results, Protein Structure, Tertiary, Apolipoprotein E4, Small Molecule Libraries, High-Throughput Screening Assays
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