Nightmares impose a substantial burden on mental health, yet their molecular determinants remain poorly understood. We employed a network pharmacology approach to identify shared protein targets of structurally diverse nightmare-inducing drugs. This approach revealed an eight-protein interactome: serotonergic (HTR2A, HTR2C, HTR1A, HTR6, HTR7), dopaminergic (DRD2, DRD3), and the potassium channel KCNH2. Protein-protein interaction networks and Gene Ontology analysis identified DRD2, HTR1A, and HTR2A as central hubs. These genes showed enrichment for neurotransmitter receptor activity and synaptic transmission. Multiple validation approaches provided initial, though not definitive, support for specificity. Negative-control drugs with lower nightmare incidence yielded a distinct intersection (ACHE and CHRM2). Permutation analysis confirmed the eight-protein composition was significantly distinct from random expectation (p = 0.0200). Experimental binding data from ChEMBL and IUPHAR/BPS confirmed high-affinity interactions for clozapine (7/8 targets) and partial support for other drugs. Analysis of nightmare-treating drugs revealed six shared proteins (HTR7, HTR2C, HTR1A, KCNH2, DRD3, DRD2). This suggests a “shared pathway perturbation” framework where directionality of modulation may determine clinical outcome. Brain expression data confirmed that all eight proteins are expressed in regions critical for sleep and emotional regulation. This framework offers a mechanistic basis for understanding nightmare susceptibility and identifies potential targets for intervention, while highlighting the need for prospective experimental validation.
Biallelic variants in the KARS1 gene, encoding lysyl-tRNA synthetase, are responsible for autosomal recessive disorders such as infantile-onset progressive leukoencephalopathy with or without deafness (LEPID; OMIM #619147) and nonsyndromic hearing loss (DFNB89). Common features of LEPID include sensorineural hearing loss, developmental delay, and leukoencephalopathy, but dysmorphic facial features and cardiac anomalies are not commonly reported. This study presents the first documented LEPID case in Iran, expanding the phenotypic and genetic spectrum of KARS1-related disorders. Whole exome sequencing (WES) was performed on a two-year-old Iranian boy from non-consanguineous parents, without a positive history of any neurologic conditions. Identified variants were validated by Sanger sequencing, with familial segregation analysis. Moreover, a literature review of 41 KARS1-related cases was conducted to compare clinical and genetic findings. WES identified two compound heterozygous KARS1 variants (NM_005548.3): c.959G > A (p.Arg320His) and c.2T > C (p.Met1Thr), previously reported and novel, respectively, both of which were classified as likely pathogenic according to the American College of Medical Genetics and Genomics criteria. The patient exhibited typical LEPID features (sensorineural hearing loss, developmental delay, seizures, leukoencephalopathy) alongside unprecedented dysmorphic facial features (e.g., synophrys, pointed triangular chin) and cardiac anomalies (pulmonary stenosis, atrial septal defect). Literature review confirmed these dysmorphic features as novel, absent in prior cases, with cardiac involvement in a limited number of patients. This study highlights a novel KARS1 variant and an expanded phenotypic spectrum of LEPID, emphasizing the role of WES in diagnosing rare disorders even in non-consanguineous marriages. These findings advocate for broader phenotypic recognition and functional studies to elucidate KARS1’s role in neurodevelopmental, craniofacial, and cardiac abnormalities.
STXBP1-related disorders (STXBP1-RD) are severe neurodevelopmental conditions associated with synaptic dysfunction, presenting an unmet medical need. P.Arg292Cys (R292C) variant is predicted to alter protein stability and affect interactions involved in synaptic vesicle fusion. This study aimed to screen for interactions between STXBP1-R292C and FDA-approved drugs and natural compounds using in silico and in vivo methods to better understand STXBP1-RD molecular defects. We employed computational structural modeling of STXBP1-R292C, screening and ranking transiently interacting drugs, followed by in vivo phenotypic assessment in a humanized STXBP1-R292C Caenorhabditis elegans (C. elegans) model, and longitudinal clinical observation in a single individual carrying the R292C variant. STXBP1-interacting FDA-approved drugs and natural compounds were identified using in silico analysis based on predicted structural effects. C. elegans locomotion features were quantified using standardized assays. Clinical outcomes were assessed using Vineland-III over 25 months. Computational modeling identified β-nicotinamide mononucleotide (NMN) as one of the lead compounds predicted to transiently stabilize the STXBP1-R292C. We further discovered that NMN was unstable in the R292C mutant STXBP1 pocket and diffused from it, whereas its metabolite the essential coenzyme, nicotinamide adenine dinucleotide (NAD+) remained stable at the binding site throughout the structural simulation. In the C. elegans model, NMN exposure partially, but significantly, modulated locomotion phenotypes toward a wild-type (WT) profile. Vineland-III assessment of a child (2–4 years old) with STXBP1-RD and motor delays showed improved motor domain scores, while other developmental domains displayed limited changes. Our study revealed an association between STXBP1-R292C/STXBP1 and NMN/NAD+ as a lead molecule. At the organismal level, NMN treatment ameliorated motor defects in humanized STXBP1-R292C C. elegans. Lastly, 2-year observations in a child carrying the same mutation suggested an NMN effect on the motor domain, toward a future well-designed clinical trial.
Phosphatidylethanolamine-Binding Protein 1 (PEBP1), also known as Raf Kinase Inhibitory Protein (RKIP), is a widely expressed, highly conserved multifunctional signaling regulator that governs the activity of central signaling pathways including Mitogen-Activated Protein Kinase (MAPK), Nuclear Factor-κB (NF-κB) and G Protein-Coupled Receptors (GPCRs). We note that recent research has uncovered its role as a critical ferroptosis regulator, showing remarkably specific expression and function across different diseases. Here we systematically review the structure and function of PEBP1, focusing on its core mechanisms as a molecular hub in various neurological disorders. We also systematically integrate PEBP1’s multiple roles across these disorders and explain its key position within neuropathological networks by coupling classical signaling pathways with the ferroptosis program. We further examine intervention strategies targeting its functional states and protein interaction networks, and then discuss its clinical translational potential as a therapeutic target, hoping to provide insights for investigations into disease-related mechanisms and the development of therapeutic strategies. Clinical trial number: not applicable.
Alzheimer’s Disease (AD) is marked by the accumulation of amyloid-beta (Aβ) plaques, neurofibrillary tangles, and neuroinflammation. Cannabidiol (CBD), the major non-psychoactive compound of Cannabis, has demonstrated significant pharmacological potential for the treatment of AD. CBD is known to act on various receptors, including cannabinoid receptor 1 (CB1R) and 2 (CB2R), and peroxisome proliferator activated receptor γ (PPARγ). However, the precise changes in receptor expression in AD, as well as following CBD treatment, remain unclear and inconsistent in literature. To address this, we performed immunohistochemistry and immunoblotting in AD-compromised brain regions, specifically the cortex and the hippocampus. In 5xFAD transgenic (TG) mice, we noted strong deposition of Aβ plaques and glial fibrillary acidic protein (GFAP)- positive astrocytes. However, following CBD treatment, Aβ plaque burden and GFAP accumulation were significantly reduced. We observed increased CB1R expression in the cortex of TG mice, in comparison to age-matched wild-type control mice. CB2R expression changes were insignificant between the two groups, however, following CBD treatment, CB2R expression was significantly increased. Interestingly, we demonstrated PPARγ co-expressed with Aβ plaques in TG mice, and following CBD treatment, PPARγ co-expression increased. Lastly, we demonstrated dysregulations of the PI3K signaling pathway observed in TG mice was ameliorated with CBD treatment. In this study, we attributed the effects of CBD on AD-related biomarkers, particularly decreased Aβ plaque load and neuroinflammation, to the alterations in CB2R and PPARγ expression.
