Journal of Molecular Neuroscience最新文献

Traumatic brain injury (TBI) is associated with complex transcriptional disturbances involving cortical suppression, neuroimmune responses, and altered neuroactive signaling. Although repetitive transcranial magnetic stimulation (rTMS) and acupuncture-related stimulation have been investigated as neuromodulatory strategies after brain injury, the transcriptomic features associated with combined rTMS and press-tack needle (PTN) stimulation remain incompletely understood. In this study, RNA sequencing was performed on prefrontal cortex tissue from Sham rats, TBI rats, and TBI rats treated with combined rTMS + PTN. Differentially expressed genes (DEGs), functional enrichment, pathway-gene interaction networks, protein-protein interaction networks, and gene set enrichment analysis (GSEA) were used to explore transcriptional alterations associated with TBI and combined intervention. TBI induced marked transcriptional changes in the prefrontal cortex, including genes enriched in neuroactive ligand-receptor interaction, calcium signaling, and injury-related pathways. Combined rTMS + PTN stimulation was associated with additional transcriptional remodeling, particularly involving immune-inflammatory pathways such as NF-κB signaling, IL-17 signaling, cytokine-cytokine receptor interaction, and Toll-like receptor signaling. Intersection analysis identified 55 shared TBI-associated and intervention-responsive genes as a candidate signature. Further PPI and GSEA analyses suggested that Nts/Ntsr1, Egr1, Drd1, and related neuroactive signaling molecules may participate in the transcriptional response to combined stimulation after TBI. These findings provide an exploratory RNA-seq-based framework for understanding NTSR1-associated neuroimmune and neuroactive molecular signatures after TBI. Targeted qRT-PCR assessment of selected neuroactive candidate genes, including Ntsrl, Egrl, and Drd1, provided complementary evidence of group-dependent expression differences in an independent animal cohort. Further protein-level and functional validation will be required to determine the causal relationships among these candidate molecules.

Postoperative delirium (POD) and Alzheimer’s disease (AD) are increasingly recognized as related neurocognitive conditions, but the aging-associated cell states that may connect them remain poorly defined. Here, we integrated two brain transcriptomic discovery datasets analyzed at single-cell/single-nucleus resolution, including a POD-related cohort (GSE291019) and an AD cohort (GSE129308), together with two independent peripheral-blood bulk transcriptomic datasets (GSE163943 and GSE63060), to identify aging-associated cellular programs and prioritize convergent molecular candidates. Across 184,168 high-quality cells, inhibitory neurons showed the most consistent aging-associated perturbation across the POD- and AD-related datasets. Re-clustering further identified three inhibitory-neuron subtypes, Inh_Neurons2, Inh_Neurons3, and Inh_Neurons5, with relatively high aging-related gene activity and preferential localization to later pseudotime states. Cross-platform integration of aging-associated inhibitory-neuron genes with a shared bulk DEG set identified four convergent candidates: RGL2, AKT1, SYK, and TNFSF13B. Among them, RGL2 emerged as the leading candidate, with the strongest downstream support concentrated in AD-related analyses. In two-sample Mendelian randomization, genetically predicted higher RGL2 expression was associated with increased AD risk, whereas the estimate for the delirium genome-wide association study proxy used for POD-related analyses was not significant. Pathway analyses further linked higher RGL2 expression to complement/coagulation and innate immune-inflammatory programs in AD-related analyses. These findings suggest a model in which POD and AD may partially intersect through aging-vulnerable inhibitory-neuron states and identify RGL2 as a prioritized candidate for downstream mechanistic investigation.

Parkinson’s disease (PD) is a progressive neurodegenerative disorder that affects over 12 million people worldwide. A central pathological feature is the accumulation of aggregated alpha-synuclein (αS) in Lewy bodies and Lewy neurites. Engineered αS variants such as 3K (E35K+E46K+E61K) and KLK (KTKEGV→KLKEGV in 6 repeats) have been shown to enhance membrane binding and aggregation propensity, contributing to cellular toxicity. To further investigate the impact of 3K and KLK on αS biology, we developed a sensitive assay in a human neuroblastoma model to assess expression levels and cytotoxicity. Relative to wild-type αS, the 3K mutant exhibited reduced steady-state expression and increased toxicity, consistent with prior reports. In contrast, the KLK mutant showed no marked change in protein expression but induced significantly higher toxicity, more than the 3K variant. Furthermore, stearoyl-CoA desaturase (SCD) inhibition partially rescued the toxicity of 3K and KLK αS, with 3K showing greater restoration of cell growth than KLK, validating the sensitivity of our live-cell assay to detect pharmacological responses. These findings underscore the utility of our assay in dissecting disease-relevant mechanisms and highlight the potential of engineered αS variants to model pathogenic features of PD. This platform offers a versatile tool for evaluating therapeutic strategies targeting αS aggregation and toxicity in PD and related synucleinopathies.

Obsessive-compulsive disorder (OCD) is linked to cortico-striato-thalamo-cortical circuit dysfunction, but molecular programs within striatal circuit nodes remain incompletely characterized. We tested whether translation, mitochondrial bioenergetics, and proteostasis show a coordinated transcriptomic signature in the OCD caudate. We reanalyzed postmortem caudate RNA-sequencing data from GSE262138 after retaining OCD cases and unaffected controls (OCD n = 27; control n = 85). Gene-level models adjusted for RNA integrity number and sex. Ranked gene statistics were used for pathway enrichment, module testing, expression-matched random-gene comparisons, sensitivity analyses, marker-score adjustment, and comparison with published OCD striatal literature. The caudate gene-level landscape showed a broad ranked-expression pattern. Pathway analysis identified recurrent lower expression of translation/ribosome, mitochondrial oxidative phosphorylation, electron-transport, and proteostasis-related terms across annotation systems. Translation/ribosome, mitochondrial/OXPHOS, and proteostasis modules showed lower average shifts relative to expression-matched random gene sets, and the direction remained consistent across sensitivity analyses. OCD caudate transcriptomic variation shows a pathway-level signature centered on translational, bioenergetic, and proteostatic programs. The results connect public postmortem caudate expression data with CSTC circuit biology and provide a focused molecular view of OCD-associated caudate dysfunction.

Migraine may involve neuroimmune-related mechanisms, but migraine-associated peripheral immune-cell states remain incompletely defined. We aimed to characterize peripheral blood immune remodeling in migraine at single-cell resolution. We reanalyzed PBMC single-cell RNA-sequencing dataset GSE269117, using migraine and vestibular migraine samples as the migraine group and healthy controls as controls. After quality control, we performed major immune-cell annotation, CD4 T-cell subclustering, pseudobulk differential-expression analysis, pathway enrichment, pseudotime reconstruction, CellChat-based communication analysis, transcription-factor activity inference and AL589693.1-focused state analysis. A total of 31,514 PBMCs from 15 retained biological samples were analyzed and annotated into six major immune-cell populations. Major lineage proportions showed no broad sample-level differences between groups, whereas sample-level analysis of 7,647 CD4 T cells indicated a redistribution of CD4 T-cell states. At the sample level, migraine samples showed enrichment of non-cytotoxic-like CD4 T cells and depletion of cytotoxic-like CD4 T cells compared with controls. CD4 pseudobulk and enrichment analyses highlighted inflammatory, estrogen-response, calcium-responsive and cAMP-related programs, with relative depletion of oxidative phosphorylation, MYC targets and translation-related pathways. Pseudotime analysis supported a shift toward earlier or less cytotoxic CD4 states. CellChat inferred fewer total interactions in migraine than in controls, especially cytotoxic-like CD4-related interactions. AL589693.1 emerged as a migraine-associated transcript, and its no-self signature marked a non-cytotoxic-skewed CD4 state that partially contributed to the CD4 imbalance. Peripheral CD4 T-cell state remodeling may represent an important feature of migraine-associated immune alterations, with AL589693.1-positive CD4 states as candidate markers for future validation.

Acute ischemic stroke is a major cause of death and disability, and post-stroke cognitive impairment remains a major clinical challenge. Cardioembolic stroke (CES), often associated with atrial fibrillation, is closely linked to cognitive decline. Increasing evidence suggests that vascular dysfunction contributes to both vascular cognitive impairment and Alzheimer’s disease (AD), but the shared molecular basis remains unclear. This study aimed to identify shared molecular signatures and candidate biomarkers linking CES and AD. Gene expression datasets were obtained from the Gene Expression Omnibus database. Differentially expressed genes were identified using limma, and disease-related gene modules were constructed using weighted gene co-expression network analysis. Functional enrichment analyses were performed to explore shared pathways. Three machine learning algorithms—LASSO, SVM-RFE, and random forest—were used to prioritize candidate genes. Receiver operating characteristic analysis, nomogram modeling, single-gene gene set enrichment analysis, and external validation were performed to evaluate the potential relevance of the identified gene. Seven overlapping genes were identified between CES and AD. Among them, SCRIB was consistently selected by the integrated feature-selection workflow and showed moderate diagnostic performance in both internal datasets. Supportive validation was observed in an independent AD dataset and in an atrial fibrillation-related surrogate validation cohort. SCRIB may represent a shared transcriptomic biomarker candidate linking CES and AD. These findings provide preliminary transcriptomic evidence for a potential molecular link between cerebrovascular pathology and neurodegeneration, but further mechanistic and clinical validation is required.

Background: The pathogenesis of ischemic stroke (IS) involves a starvation response (SR); however, the mechanistic link between the two remains unclear. Therefore, it is imperative to identify SR-related gene (SRRG) biomarkers involved in IS. Methods: IS datasets were obtained from the Gene Expression Omnibus, and SRRGs were acquired from the Molecular Signatures Database. Biomarkers were screened using differential expression analysis, machine learning, and expression validation. Subsequent analyses included immune infiltration, gene set enrichment analysis (GSEA), drug prediction, and single‑cell RNA sequencing (scRNA‑seq). The middle cerebral artery occlusion mouse model and oxygen-glucose deprivation/reoxygenation (OGD/R)-treated BV2 cells were used. PAK2/Iba1 co-expression was assessed using immunofluorescence double-staining. The effects of PAK2 knockdown on cell proliferation, apoptosis, and inflammation were assessed using functional assays. Results: Two biomarkers, PAK2 and lysosome-associated membrane protein 2 (LAMP2), were identified and used to construct a diagnostic model that demonstrated good performance. Immune infiltration analysis revealed changes in 33 immune cell types. GSEA revealed that biomarker‑related genes were enriched in different pathways, including NOD‑like receptor and T‑cell receptor signaling pathways. Drug prediction analysis identified 48 potential agents targeting PAK2 and 35 agents targeting LAMP2. Microglia were identified as a key cell subtype using scRNA‑seq analysis. PAK2 mRNA was significantly upregulated, and LAMP2 mRNA was downregulated in mouse ischemic penumbra; immunofluorescence confirmed PAK2/Iba1 co-localization. In vitro, PAK2 knockdown suppressed apoptosis, promoted proliferation, and reduced inflammatory cytokine levels in OGD/R-injured BV2 cells. Conclusion: In this study, two SR‑derived biomarkers were identified for IS, and a diagnostic model was established, providing new insights into IS diagnosis and therapeutic development.

Historically the development of amyloid-β plaques and tau neurofibrillary tangles have been used as the hallmarks of Alzheimer’s disease (AD). However, there is increasing evidence suggests that these pathological hallmarks are secondary to deeper metabolic defect in the brain. AD is progressively being recognized as a metabolic synaptic disorder characterized by insulin resistance, impaired cellular energy homeostasis, mitochondrial dysfunction, and synaptic degeneration. Synaptic plasticity is closely linked to the insulin-sensitive system of glucose utilization, mitochondrial activity, and local protein synthesis that render the synapses highly sensitive to the malfunction of the metabolic system. Recent discoveries highlight the important role of exosomes in mediating communication between neural cells by transferring regulatory miRNAs across neuronal networks. Exosomal miRNAs regulate insulin signaling, synaptic gene expression, mitochondrial function, and neuroinflammation. In AD, exosomal miRNA profiles are significantly altered, with enrichment of miR-29, miR-34a, miR-146a, and miR-21, alongside depletion of synapse-supporting miR-132. These changes contribute to insulin resistance, impaired glucose transporter trafficking, dendritic spine destabilization, and reduced expression of synaptic proteins such as PSD-95 and synaptophysin, ultimately leading to cognitive decline. Importantly, neuron-derived exosomes can cross the blood–brain barrier, making their miRNA cargo promising biomarkers and therapeutic targets for early AD diagnosis and precision treatment.

Graphical Abstract

Graphical abstract illustrates Alzheimer’s disease as a disorder of corrupted exosome-mediated metabolic communication.