Bhattrai, A. | Search bioRxiv

bioRxiv2026

Addition of humanized APP to humanized APOE mouse model reduces brain size and increases the ratio of cortical representation

INTRODUCTION: Age, Apolipoprotein E4 (APOE4) genotype, and biological sex are major risk factors for late-onset Alzheimer's disease (LOAD), and preclinical mouse models enable controlled investigation of these factors. To date, humanized APOE4 has not recapitulated LOAD-relevant brain volume phenotypes. Given the central role of amyloid precursor protein (APP) in LOAD pathogenesis, incorporating humanized APP (hAPP) alongside humanized APOE (hAPOE) may therefore improve translational modeling of structural brain changes characterized by neuroimaging. METHODS: Aged mice (mean age = 23.25 months) carrying murine (m) or humanized (h) APP and either murine Apoe or hAPOE3/3 (hAPOE3-HOM), hAPOE3/4 (hAPOE4-HET), or hAPOE4/4 (hAPOE4-HOM) underwent in-skull ex vivo volumetric MRI. Regional volumes were quantified in absolute terms and relative to total brain volume (TBV). Linear models included APP type, APOE genotype, and sex, with FDR correction applied within contrasts. RESULTS: Brain volumes were primarily determined by APP background, with hAPP globally reducing total and regional volumes relative to mAPP mice. Across hAPP models, hAPOE4-HOM exhibited the greatest brain-wide reductions, which was mitigated by a single hAPOE3 allele. In contrast, mouse APP exerted modest effect in hAPOE, with hAPOE4 carriers exhibiting greater total volume without regional specificity. After TBV adjustment, hAPP mice exhibited subcortical vulnerability with relative cortical preservation. Females exhibited larger brain volumes than males, independent of APP or APOE genotype. DISCUSSION: hAPP induces distinctly smaller brain volumes in this humanized APOE knock-in model, and hAPOE4 homozygosity amplifies that effect, indicating genotype-dependent susceptibility. Because this study is cross-sectional and lacks histopathological confirmation, volume differences may reflect developmental or constitutive effects rather than neurodegeneration. Humanized APP and APOE are therefore necessary but not sufficient to recapitulate the complete volumetric signature of established LOAD. Longitudinal and histological studies are required to determine whether these differences reflect a prodromal trajectory or a developmental effect.

neuroscience↗

bioRxiv2025

Humanized APOE mouse brain volume increases over age irrespective of sex and APOE genotype: Implications for translational validity to the human

Translational validity of mouse models of human aging and late-onset Alzheimers disease (LOAD) risk are essential for both fundamental mechanistic science and therapeutic development. Given that the strongest risk factors for LOAD are age, female sex, and APOE-{varepsilon}4 carriership, models must reflect these biological variables and disease phenotypes. The use of mouse models with humanized APOE (hAPOE) is a key strategy to advance translational validity. To initially address translational validity of the hAPOE mouse model, we conducted ex-vivo magnetic resonance imaging analysis of brain volumes in male and female mice across APOE genotypes ({varepsilon}3/{varepsilon}3, {varepsilon}3/{varepsilon}4, {varepsilon}4/{varepsilon}4) and ages corresponding to a human lifespan of approximately 30-70 years (6-25 months in mice). The primary outcomes indicated that total MRI brain volume increased with age (an average of 2.12mm3 per month), irrespective of sex or APOE genotype. Additionally, APOE-{varepsilon}4 carriers had greater total brain volumes than non-carriers. No sex differences were observed in total brain volume. Voxelwise analyses revealed a pattern of localized morphometric changes independent of differences in total brain volume. Age-related volumetric increases were distributed across subcortical regions (e.g., thalami, hippocampi), while age-related decreases were evident across cortical regions, notably the bilateral parieto-temporal and frontal lobes. Additionally, sex differences were evident after controlling for total brain volume, with females showing greater cortex-dominant volumes while males showed a pattern of greater volumes in regions including cerebellar cortices, olfactory bulbs, and striata. No genotypic effects were observed in the voxelwise analysis after correcting for multiple comparisons, suggesting that APOE genotype does not drive localized volume differences independent of total brain volume. These findings indicate that, at the MRI level of analysis, this humanized APOE mouse model does not recapitulate the volumetric atrophy typically seen in human brain aging and Alzheimers disease. The results suggest that humanized APOE alone is insufficient to induce the LOAD atrophy phenotype. This model may better serve as a platform for studying vulnerable aging rather than a primary model for progressive neurodegenerative atrophy.

neuroscience↗