Alzheimer’s disease (AD) is one of the most prevalent neurodegenerative diseases and a major contributor to dementia. Although the cause of this condition has been identified long ago as aberrant aggregations of amyloid and tau proteins, effective therapies for it remain elusive. The complexities of drug development for AD treatment are often compounded by the impermeable blood–brain barrier and low-yield brain delivery. In addition, the use of high drug concentrations to overcome this challenge may entail side effects. To address these challenges and enhance the precision of delivery into brain regions affected by amyloid aggregation, we proposed a transferrin-conjugated nanoparticle-based drug delivery system. The transferrin-conjugated melittin-loaded L-arginine-coated iron oxide nanoparticles (Tf-MeLioNs) developed in this study successfully mitigated melittin-induced cytotoxicity and hemolysis in the cell culture system. In the 5XFAD mouse brain, Tf-MeLioNs remarkably reduced amyloid plaque accumulation, particularly in the hippocampus. This study suggested Tf-LioNs as a potential drug delivery platform and Tf-MeLioNs as a candidate for therapeutic drug targeting of amyloid plaques in AD. These findings provide a foundation for further exploration and advancement in AD therapeutics.

Alzheimer’s disease (AD) is characterized by the accumulation of amyloid β (Aβ) plaques in the brain, leading to cognitive impairment and other clinical symptoms. The 5XFAD mouse model is commonly used in AD research because it expresses five human transgenes that result in the accumulation of Aβ plaques and cognitive decline at a relatively early age. Behavioral experiments are frequently conducted using this model; however, the effect size has not yet been reported. In this study, we examined basic cognition and locomotion in 5XFAD mice with a C57BL6/J background (5XFAD-J) at 6 months of age, a period in which impairments of cognitive function and locomotion are commonly observed. We analyzed the effect sizes of cognitive and locomotive experiments in the 5XFAD mice compared with those in the wild-type mice. Our results suggest that for long-term memory analysis, the novel object recognition test (p = 0.013, effect size 1.24) required a sample size of at least 12 to obtain meaningful results. Moreover, analysis of general locomotion over total distance with the Laboratory Animal Behavior Observation, Registration and Analysis System (LABORAS) test during the dark phase (p = 0.007, effect size −1.37) needed a sample size of 10 for a statistical power (1-β) of 0.8. In conclusion, we can conduct more ethical and scientifically rigorous animal experiments using 5XFAD mice based on the effect and sample sizes suggested in this study.

The typical pathological features of Alzheimer’s disease (AD) are the accumulation of amyloid plaques in the brain and reactivity of glial cells such as astrocytes and microglia. Clinically, the development of AD and obesity are known to be correlated. In this study, we analyzed the changes in AD pathological characteristics in 5XFAD mice after obesity induction through a high-fat diet (HFD). Surprisingly, high-density lipoprotein and apolipoprotein AI (APOA-I) serum levels were increased without low-density lipoprotein alteration in both HFD groups. The reactivity of astrocytes and microglia in the dentate gyrus of the hippocampus and fornix of the hypothalamus in 5XFAD mice was decreased in the transgenic (TG)-HFD high group. Finally, the accumulation of amyloid plaques in the dentate gyrus region of the hippocampus was also significantly decreased in the TG-HFD high group. These results suggest that increased high-density lipoprotein level, especially with increased APOA-I serum level, alleviates the pathological features of AD and could be a new potential therapeutic strategy for AD treatment.

Alzheimer's disease (AD) progression involves systemic network transitions. To capture these using ROSMAP bulk RNA–seq (n=624), we focused on the geometry of the covariance structure, performing a Riemannian (Log–Euclidean) analysis of stage-wise covariance matrices as points on the manifold of symmetric positive-definite (SPD) matrices. On the SPD manifold the three stages were non–collinear: geodesic distances were non–uniform and MCI was displaced from the NCI–AD chord, while the von Neumann entropy of the covariance structure dipped at MCI (S = 2.760, 2.639, 2.647 for normal cognitive intact NCI, mild cognitive impairment MCI, AD) and the path-curvature profile reached a minimum there — together identifying MCI as a saddle/bifurcation state. The differential covariance spectrum (CAD – CNCI) separated AD–amplified ("structural collapse") from AD–suppressed ("protective loss") modes. Ultimately, second–order statistics analyzed through Riemannian geometry, rather than Euclidean summaries, reveal AD progression structure invisible to mean-level analysis.

Synucleinopathies, including Parkinson's disease (PD) and Lewy body dementia (LBD), are characterized by misfolding and aggregation of α-synuclein (α-syn) into Lewy bodies, a key pathological hallmark. These disorders display region-specific vulnerability, with PD affecting the nigrostriatal dopaminergic system and LBD involving the neocortex. However, the molecular basis of selective vulnerability, particularly hippocampal involvement, remains poorly understood. We aimed to determine whether the G2-3 α-synuclein transgenic mouse model recapitulates region-specific vulnerabilities observed in human synucleinopathies and to identify molecular and cellular mechanisms underlying region-specific pathology. Six-month-old G2-3 mice and wild-type (WT) littermates underwent spatial transcriptomics profiling, followed by Gene Ontology (GO) and KEGG enrichment analyses. Key findings were validated using immunohistochemistry and cell-type deconvolution. Spatial transcriptomics revealed region-specific transcriptomic alterations in the G2-3 synucleinopathy mouse brain, with the hippocampus-particularly the DG and CA subfields-showing the most pronounced changes. GO analysis highlighted enrichment of synapse-related processes, and immunohistochemistry confirmed reduced and mislocalized PSD95 with glial colocalization. KEGG analysis further identified MAPK signaling activation, validated by increased neuronal p-ERK, underscoring hippocampus-specific molecular and structural vulnerabilities under α-syn pathology. Our study demonstrates that the hippocampus is a selectively vulnerable region in the G2-3 synucleinopathy mouse model, showing both transcriptional and structural synaptic alterations. Spatial transcriptomics revealed hippocampal enrichment of synapse-related processes and MAPK signaling, which were corroborated by immunohistochemical evidence of PSD95 mislocalization and neuronal p-ERK activation. These results provide mechanistic insights into hippocampal vulnerability and highlight spatial transcriptomics as a powerful tool to uncover region-specific disease mechanisms in synucleinopathies.

Cerebrospinal fluid (CSF) flow and the glymphatic system are essential for maintaining brain homeostasis, yet the precise mechanisms driving their profound failure in Alzheimer’s disease (AD) remain largely elusive. Here, we reveal that macroscopic CSF propagates dorsally via the perivascular spaces (PAS) of major ventral arteries—the Circle of Willis (CW), middle cerebral artery (MCA), and olfactory artery (OlfA)—which undergo severe structural degradation in an AD mouse model. High-resolution imaging of the dorsal cortex demonstrates that this structural collapse induces a severe influx blockade; in AD, CSF tracers form massive aggregates within the PAS, physically decoupling the gliovascular interface, while tracers entering the parenchyma are pathologically sequestered by amyloid plaques. Furthermore, we identify an active, cholesterol-mediated clearance mechanism responding to exogenous CSF tracer infiltration. While healthy brains rapidly synthesize and efficiently clear this cholesterol via vascular efflux within 120 minutes, the AD model exhibits blunted cholesterol mobilization and a complete failure of capillary efflux. This terminal bottleneck forces uncleared cholesterol to retrogradely accumulate into pathological aggregates within perivascular pericytes and neuronal somata. Collectively, our findings demonstrate that AD dismantles the entire CSF recirculation loop—from macroscopic ventral entry to terminal microvascular efflux—highlighting pericytic and neuronal lipidosis as fundamental drivers of brain fluid stagnation.