Search Results (108)

Epidemiological studies have identified the developing brain as a target of concern for polychlorinated biphenyls (PCBs). In animal models, behavioral deficits caused by developmental exposure to PCBs have been associated with altered patterns of dendritic arborization in functionally relevant brain regions. In vitro studies revealed that PCB 95 promoted dendritic growth in primary rat hippocampal neuron–glia co-cultures via ryanodine receptor 1 (RYR1)-dependent Ca²⁺ signaling. However, it is not yet known whether RYR1 dysregulation contributes to disruption of dendritic morphogenesis in the intact developing brain or whether PCB 95 affects translationally relevant behavioral endpoints in juvenile animals. To address these data gaps, we assessed Morris water maze (MWM) performance and dendritic arborization of hippocampal CA1 pyramidal neurons in C57BL/6 mice heterozygous for the human R163C-RYR1 gain-of-function mutation (HET) and congenic wildtype (WT) littermates exposed to vehicle or PCB 95 at 0.1, 1.0, or 6.0 mg/kg/d in the dam’s diet from conception through weaning. MWM performance was not altered in HET vehicle controls relative to WT vehicle controls; however, compared to genotype-matched vehicle controls, spatial learning was impaired in WT and HET male and female weanlings in the 1.0 mg/kg/d PCB 95 dose group and WT males in the 6.0 mg/kg/d PCB 95 dose group. Spatial memory was altered only in WT females exposed to 1.0 or 6.0 mg/kg/d PCB 95. Sholl analyses of Golgi-stained hippocampal neurons in male and female WT and HET weanlings from the 1.0 mg/kg/d PCB 95 and vehicle groups indicated that relative to WT vehicle controls, basal dendritic arborization was significantly increased in HET vehicle controls and in PCB 95-exposed WT weanlings; however, PCB 95 did not alter basal dendritic growth in HET weanlings relative to genotype-matched vehicle controls. PCB 95 significantly reduced training-induced dendritic arborization in WT but not HET weanlings. Measurement of tritiated ryanodine ([³H]Ry) binding in cortical tissue from these same animals revealed interactions between genotype, PCB 95 exposure, and dose that altered [³H]Ry binding relative to WT vehicle controls. Quantitative analyses confirmed a dose-dependent increase in PCB tissue burden that was not significantly altered by genotype, MWM training, or sex. Serum levels of progesterone, estradiol, cortisol, and thyroid hormone (TH) and brain transcript levels of TH-responsive genes were not significantly altered by genotype or PCB 95 dose. These data demonstrate that PCB 95 caused behavioral deficits coincident with altered patterns of basal and training-induced dendritic arborization. Furthermore, behavioral and dendritic responses to PCB 95 were altered by expression of the human R163C-RYR1 gain-of-function mutation, supporting the involvement of RYR1-dependent mechanisms in PCB 95 DNT in vivo. Full article

Background: Duchenne muscular dystrophy (DMD) is a monogenic disorder caused by dystrophin deficiency, but its downstream molecular consequences involve complex and interconnected regulatory networks. Methods: In this study, we performed a cross-species integrative transcriptomic analysis of skeletal muscle RNA-seq datasets from humans, mice, and rats to identify conserved molecular signatures of DMD. Results: Differential expression analysis identified 1367 shared differentially expressed genes (DEGs) across the three datasets, of which 62 showed concordant directionality in all species. After ortholog filtering, which excluded five genes lacking confirmed one-to-one orthologs across all three species, 57 conserved genes were retained as the conserved DMD-associated gene (DAG) set. Functional enrichment (GO/KEGG) was performed on the cross-species shared DEG set, and the 57 conserved genes were subsequently mapped onto these enrichment outputs. The DAG-containing categories included focal adhesion, apoptosis, and calcium signaling, which remained significant after Benjamini–Hochberg correction, together with nominally enriched Rap1 signaling. Integration of circRNA, miRNA, and mRNA data further identified a candidate, computationally inferred hub-dominated ceRNA network involving DMD, RYR3, KLF4, ID1, TIMP4, and CASP7. Conclusions: Together, these findings support a model in which conserved core molecular programs are superimposed on species- and context-dependent transcriptional responses, prioritizing the conserved genes and candidate circRNA-related network components for future mechanistic and translational studies. Full article

Background/Objectives: Malignant hyperthermia (MH) is a life-threatening pharmacogenetic disorder of skeletal muscle primarily associated with pathogenic variants in RYR1; however, a substantial proportion of MH-susceptible individuals lack identifiable variants in known genes. This study aimed to identify novel genetic contributors in Ca²⁺-induced Ca²⁺ release (CICR)-positive patients without RYR1 variants and to evaluate their functional relevance. Methods: Among 29 CICR-positive individuals without pathogenic variants identified by gene panel testing, five patients underwent whole-exome sequencing (WES). In one family, heterozygous variants in ANO5 (p.Arg547Gln) and ORAI3 (p.Arg287Cys) were identified. To evaluate their functional significance, intracellular Ca²⁺ dynamics were analyzed in primary myotubes derived from Case 165, an affected individual carrying both variants, and compared with those from CICR-negative controls and CICR-positive patients harboring RYR1 variants. Results: Myotubes derived from Case 165 demonstrated enhanced sensitivity to caffeine and 4-chloro-m-cresol, elevated resting intracellular Ca²⁺ levels, and greater Ca²⁺ reduction under Ca²⁺-free conditions compared with CICR-negative controls, resembling the phenotype observed in the RYR1 variant group. In contrast, dantrolene-induced Ca²⁺ reduction was significantly greater only in the RYR1 variant group. Conclusions: These findings suggest that ANO5 and ORAI3 variants may contribute to abnormal Ca²⁺ regulation in MH-susceptible individuals without RYR1 mutations and highlight the importance of combining genomic analysis with functional validation to identify novel genetic contributors to MH susceptibility. Full article

Red yeast rice (RYR) food supplements are widely used for cholesterol management due to their monacolin content, particularly monacolin K (MK), which shares the same chemical structure as lovastatin, a compound associated with several side effects. The European Food Safety Authority (EFSA) highlighted uncertainties regarding the compositional variability, characterization of bioactive constituents, and safety of RYR products, concluding that a safe level of monacolin intake could not be established. In this study, twenty-seven commercial semi-processed RYR ingredients for food supplements manufacturing were investigated through a combined chemical and biological approach to characterize their composition and evaluate potential safety concerns. Targeted LC-MS/MS analyses revealed substantial variability in MK, monacolin hydroxy acid (MKA), and dehydromonacolin K (DMK) levels among products. Exploratory qualitative untargeted metabolomics enabled the putative annotation of multiple secondary metabolites, including azaphilones and polyketides, highlighting differences in chemical profiles. Correlation analyses indicated that DMK content was associated with pigment distribution and cytotoxic responses in human hepatocellular carcinoma cells and primary human skeletal muscle cells, whereas MK and MKA showed no significant association with toxicity under the tested conditions. Exploratory proteomic profiling suggested distinct protein-abundance patterns between RYR- and simvastatin-treated cells, providing hypothesis-generating information on functional annotations associated with these patterns. These findings emphasize the importance of comprehensive chemical characterization and quality control strategies considering both monacolins and secondary metabolites to improve the safety assessment of RYR supplements. Full article

Ryanodine receptor-like (RyR-like) is a key endoplasmic reticulum (ER) Ca²⁺ release channel governing intracellular Ca²⁺ homeostasis and cellular stress responses in invertebrates. However, its function in bivalves under high-temperature stress remains unclear. In the present study, one RyR-like was identified from Yesso scallop Patinopecten yessoensis (PyRyR-like). Its function in regulating intracellular Ca²⁺, IRE1α-mediated unfolded protein response (UPR) and apoptosis in the mantle after high-temperature (25 °C) treatment was investigated using molecular cloning, qRT-PCR, Western blot, pull-down assay, cellular calcium imaging, TUNEL and histology assays; High temperature treatment significantly increased intracellular Ca²⁺ content at 1 and 6 h (p < 0.05), but decreased it at 3, 12 and 24 h (p < 0.05); meanwhile, the cAMP level, PyPKA activity, mRNA expression level of PyRyR-like, and protein expression levels of PyFKBP12 and PyGRP78 were significantly increased at different times. However, high temperature did not affect the expression level of PyNVL and PyXBP1(S). The SPRY and RYR domains of PyRyR-like separately interacted with PyFKBP12 and PyPKA. Moreover, RyR antagonist Dantrolene reversed high-temperature-induced alterations in Ca concentration, PKA activity, and core UPR- and apoptosis-related molecules, and suppressed Caspase-3 activity. These findings suggest that PyRyR-like plays an important role in the high-temperature response of scallops by regulating intracellular Ca²⁺ homeostasis and mediating UPR activation and apoptosis, providing new insight into the molecular mechanism underlying scallop adaptation to high temperature. Full article

Ventricular remodeling occurring in heart failure (HF) involves structural disarray of the sarcolemma T-tubule (TT)–sarcoplasmic reticulum (SR) dyad junctions, thereby disrupting the close apposition of L-type Ca²⁺ channels (Ca_V1.2) with ryanodine receptors (RyR2) that trigger SR Ca²⁺ release and myofilament contraction. In a rat ischemic heart failure model expressing low thyroid hormone (TH) function, we used 3D stochastic optical reconstruction microscopy (STORM) to image RyR2 clusters with Ca_V1.2 channels, and the associated protein junctophilin-2 (Jph2). We tested whether treatment with T3, the biologically active form of TH, throughout progression of the disease would preserve T-tubule structure and dyadic ion channel organization. Confocal microscopy of isolated cardiomyocytes (CMs) stained with ANEPPS membrane dye showed significantly decreased TT density in diseased CMs while T3 treatment attenuated TT disorganization. 3D STORM images of dyadic ion channels labeled with fluorescent-tagged antibodies to RyR-Dylight550, Jph-CF647 and Ca_V1.2/IgG-Dylight488 were captured. A density-based algorithm defined RyR2 clusters, and a 400 nm spherical 3D volume of interest around each RyR2 cluster’s centroid determined the number of Ca_V1.2 and Jph2 localizations associated with each RyR2 cluster. Analysis revealed significant reduction in RyR2 cluster size and number with reduced co-localized Jph2 in failing CMs. T3 treatment increased RyR2 cluster numbers and cluster volumes albeit non-significantly, with increased co-clustering of Jph2. The number of Ca_V1.2 co-localized with RyR2 clusters trended lower in the failing CMs. These results support maintaining TH homeostasis in optimizing the nanoscale organization of Ca²⁺ ion channels in triggering Ca²⁺ release and myofibrillar contraction in patients with heart disease. Full article

Calcium sparks can arise as both voltage-dependent and voltage-independent ligand-activated release events in amphibian skeletal muscle. To assess their gating behavior, calcium sparks were recorded from intact frog skeletal muscle fibers using high-temporal-resolution confocal microscopy (line scans: 15 and 50 µs/line). Sparks were triggered by 1 mmol/L caffeine to open ryanodine receptors (RyRs) or by subthreshold depolarization to a −65 mV membrane potential to activate dihydropyridine receptors (DHPRs). Both treatments increased the frequency of sparks and altered their morphology. The sparks were significantly greater after caffeine treatment than in depolarized cells. The signal mass of sparks (i.e., the amount of calcium released) resembled the amplitude in shape. Additionally, the calcium release flux followed a staggered function during the activation of sparks. The detailed analysis of the sparks’ time profile revealed that the events were activated in a stepwise manner. The average step size (in F/F₀; 0.071 ± 0.003) remained constant regardless of the scanning speed. The number of steps during the activation of sparks followed a linear function based on the spark’s amplitude. Our results suggest that the activation of neighboring release units may occur sequentially, and the amplitude of the sparks depends linearly on the number of activated RyR channels. Full article

Genomic sequencing can reveal potentially life-threatening clinically actionable secondary findings in healthy individuals. Little is known about the spectrum and frequency of secondary findings in healthy people in Russia. Here, we analyzed whole-genome sequences of 42,826 healthy volunteers from urban populations across Russia, focusing on known pathogenic and likely pathogenic variants of 81 genes associated with treatable or preventable monogenic diseases listed in the American College of Medical Genetics and Genomics’ Secondary Findings recommendations (ACMG SF v3.2). Based on the ClinVar 20250421 version, secondary findings were detected in 1186 (2.76%) participants. Cancer phenotypes were the most common category of secondary findings present in 565 (1.32%) participants, followed by cardiovascular phenotypes (454 individuals, 1.05%). Genes harboring the most frequent variants were BRCA1 (151 variants), BRCA2 (100), RYR1 (93), and LDLR (71). In addition, we found 238 potential loss-of-function variants in dominant ACMG SF v3.2 list genes in 280 (0.65%) participants, which, if confirmed by orthogonal methods, could increase the frequency of secondary findings to 3.41%. A study of such depth and scale was performed for the first time in the Russian population. Full article

Type-1 ryanodine receptor (RyR1) is essential for skeletal muscle contraction. This Ca²⁺ release channel is expressed in cardiac myocytes; however, its function remains elusive. Cardiac-specific RyR1 overexpression (OE) mice were generated under the cardiac-specific Myh6 promoter. Cardiac hypertrophy (CH), cardiac functions, and mechanistic changes in RyR1 OE and control (wildtype, WT) mice were assessed using hematoxylin and eosin staining, echocardiography, electrocardiogram, quantitative RT-PCR, Western blotting, [³H]-ryanodine binding assay, confocal microscope, ROS dye Amplex Red and 2′,7′-dichlorofluorescein diacetate. RyR1 OE mice had increased whole heart, left ventricular weight, and left ventricular wall thickness, but decreased cardiac output and stroke volume, thereby presenting CH and heart failure (HF). CH markers like ANF, BNF, and aSKA mRNAs were increased in RyR1 OE heart. RyR1, but not RyR2 or RyR3, expression was increased in the RyR1 OE mouse heart. Similar results were found in mice with TAC-induced CH. RyR1, but not RyR2 mRNA, was increased in cardiac muscle from dogs and humans with CH and/or HF. Maximum [³H]-ryanodine binding was increased, whereas the binding dissociation constant decreased in left ventricular cardiomyocytes from RyR1 OE mice. RyR2-dependent Ca²⁺ sparks were increased, which was blocked by riluzole, a small molecule known to inhibit RyR2. Consistently, ROS was remarkably increased in RyR1 OE cardiac cells. We first generated cardiac-specific RyR1 OE mice; these mice had CH, HF, and increased RyR1 expression with no RyR2 or RyR3 alteration. Similar changes were observed in mice, dogs, and humans with CH and HF. Increased mitochondrial ROS-dependent RyR2 Ca²⁺ release was essential for RyR1-induced CH and HF. Full article

Objective: This study aims to investigate the protective effect of Shengmai San (SMS) against high-glucose (HG)-induced injury in neonatal rat ventricular myocytes (NRVMs) and to elucidate the underlying pharmacological molecular mechanisms. We hypothesize that SMS ameliorates HG-induced calcium homeostasis imbalance in NRVMs by improving mitochondrial energy metabolism disorder, and this protective effect is associated with the downregulation of oxidized and phosphorylated CaMKII expression to inhibit CaMKII signaling pathway overactivation. Herein, we verify this hypothesis by assessing mitochondrial function, calcium transients, sarcoplasmic reticulum (SR) calcium handling and CaMKII phosphorylation levels in NRVMs. Methods: First, ultra-high performance liquid chromatography–high resolution mass spectrometry was used to identify the chemical components of SMS to clarify its material basis. Primary NRVMs were then cultured under low-glucose (LG) or HG conditions, with 2% SMS-medicated serum (SMS-MS) as the experimental intervention, and NAC (ROS scavenger) and KN93 (CaMKII inhibitor) as positive controls. Following intervention, we sequentially detected key indicators corresponding to the proposed pathological pathway: intracellular reactive oxygen species (ROS) levels (oxidative stress), mitochondrial ROS, mitochondrial function indices including oxygen consumption rate (OCR) (energy metabolism), calcium transients and diastolic intracellular free calcium concentration (global calcium homeostasis), sarcoplasmic reticulum (SR) calcium leak (calcium handling disorder), and, finally, the phosphorylation, oxidation levels of CaMKII and RyR2 phosphorylation (Ser2814) (p-RyR2) (key regulatory pathway) via Western blot to systematically elucidate the mechanistic link between SMS intervention and HG-induced NRVM injury. Results: Quantitative analysis revealed that high-glucose (HG) induction significantly reduced calcium transient amplitude and prolonged the decay time constant (tau) in NRVMs at 72 h (p < 0.01 vs. LG), with these parameters normalizing by 120 h—an effect indicative of a compensatory adaptive response. The 2%SMS-MS markedly ameliorated HG-induced calcium transient abnormalities at 72 h (p < 0.01 vs. HG). Additionally, 2%SMS-MS significantly enhanced mitochondrial basal oxygen consumption rate, spare respiratory capacity, ATP production, and maximal respiration in HG-exposed NRVMs (p < 0.01 vs. HG). SMS also significantly reduced intracellular reactive oxygen species (ROS) levels (p < 0.01 vs. HG), mitochondrial ROS levels (p < 0.01 vs. HG), diastolic intracellular free calcium concentration (p < 0.01 vs. HG), and SR calcium leak (p < 0.05 vs. HG). Western blot analysis revealed that 2%SMS-MS intervention effectively downregulated the expression of oxidized CaMKII (Ox-CaMKII) (p < 0.01 vs. HG), phosphorylated CaMKII (p-CaMKII) (p < 0.01 vs. HG), and RyR2 phosphorylation (Ser2814) (p < 0.05 vs. HG), which may be the potential mechanism in maintaining calcium homeostasis in HG-induced NRVMs. Conclusions: This study suggests that SMS enhances mitochondrial energy metabolism and exerts a protective effect against high-glucose-induced calcium homeostasis imbalance in NRVMs, which supports our proposed hypothesis. Its potential mechanism indicates that the protective effects of SMS are associated with its ability to downregulate the expression of oxidized and phosphorylated CaMKII. These findings highlight SMS as a potential therapeutic candidate for alleviating HG-related myocardial injury and provide evidence for its application in the prevention of early diabetic cardiomyopathy. Full article

Red yeast rice (RYR), a traditional fermented product obtained via rice fermentation with Monascus purpureus, has a millennia-long history of culinary and medicinal use in East Asia and has gained global attention as a prominent functional food ingredient for its well-recognized cholesterol-lowering properties. This review is driven by one core question: How can the dual challenges of standardizing key bioactive constituents, particularly monacolin K (MK), while eliminating the mycotoxin citrinin be addressed through biotechnological and analytical advances? This narrative review consolidates the latest research progress on RYR-derived bioactive compounds, with a specific focus on their production optimization, multifaceted health-promoting potentials, safety regulation, and application prospects in health food development. We elaborate on key advances in fermentation biotechnology and strain engineering for enhancing the yield of the core lipid-lowering component MK while eliminating the nephrotoxic mycotoxin citrinin, and comprehensively summarize the synergistic bioactivities of RYR metabolites beyond MK. The current applications of RYR in functional foods, dietary supplements, and traditional fermented products are detailed, alongside a comparison of the divergent regulatory frameworks for RYR across major global markets. Finally, we identify critical bottlenecks restricting RYR industrialization, including extreme inter-product heterogeneity and global regulatory fragmentation, and propose evidence-based future research directions to facilitate the development of safe, standardized, and effective RYR-based health foods. Full article

Cardiac Ca²⁺ handling is critical for excitation–contraction coupling (ECC), with the ryanodine receptor type 2 (RyR2) serving as the key sarcoplasmic reticulum (SR) Ca²⁺ release channel in cardiomyocytes. The dysfunction of RyR2 is linked to fatal cardiac arrhythmias, including heart failure (HF) and catecholaminergic polymorphic ventricular tachycardia (CPVT). This review aims to elucidate the structural basis of RyR2, its core role in cardiac ECC and Ca²⁺ homeostasis, and the regulatory mechanisms of key modulators on its activity. By integrating recent high-resolution cryo-EM structural analyses with molecular and cellular studies on RyR2 regulation, as well as clinical evidence of RyR2 mutations in arrhythmogenic heart diseases, we provide a comprehensive overview of the field. Cryo-EM has unraveled RyR2’s gating mechanisms, ligand-binding sites, and structural features. Functionally, RyR2 mediates calcium-induced calcium release (CICR) and maintains Ca²⁺ homeostasis through coordination with SERCA2a and NCX. Key modulators (CaM, FKBP12.6, and PKA/CaMKII) and disease-linked mutations regulate RyR2 activity through distinct pathways, with defective RyR2 leading to store-overload-induced Ca²⁺ release (SOICR) and arrhythmias. Furthermore, reactive oxygen species (ROS) can induce RyR2 oxidation, establishing a pathological Ca²⁺ leak-ROS cycle in heart disease. In conclusion, RyR2 is a pivotal sensor of myocardial function, with its structural and regulatory mechanisms now well-characterized by recent studies. However, the effects of numerous RyR2 mutations remain unclear, and deeper mechanistic insights will lay a key foundation for developing novel therapies against RyR2-related cardiac diseases. Full article

To address the limited functional diversity of traditional kombucha, this study utilized red yeast rice (RYR) as an alternative substrate and prepared three samples: black tea kombucha (KBT), black tea-red yeast rice mixed kombucha (KBL, at a 1:1 ratio), and red yeast rice kombucha (KRY). After 9 days of fermentation, KRY exhibited the lowest pH, the highest total acidity, and notable sugar metabolic activity. It exhibited in vitro inhibition rates of 82.8%, 78.2%, 70.3%, and 76.9% against cholesterol esterase, pancreatic lipase, α-glucosidase, and α-amylase, respectively, indicating potential hypoglycemic and hypolipidemic activities. In contrast, KBT maintained the strongest antioxidant capacity, with scavenging rates exceeding 90% against both 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS). A total of 72 volatile flavor compounds (VFCs) were identified, with 7 key compounds enriched in KRY, which enhanced its sensory acceptance and received the highest scores in color, clarity, and aroma. Microbial community analysis revealed the post-fermentation dominance of Komagataeibacter, Acetobacter, and Saccharomyces, which correlated positively with key VFCs. These findings indicate that RYR as a substrate enhances functional microbial growth, sugar metabolism, organic acid production, flavor enrichment, and in vitro inhibitory activity of enzymes associated with hypoglycemic and hypolipidemic effects. Full article

Gestational diabetes mellitus (GDM) develops silently during early pregnancy, yet its earliest circulating molecular signatures remain poorly defined. In this exploratory biomarker study, we characterized first-trimester circulating microRNA (miRNAs) associated with later GDM using a pool-based small RNA sequencing approach. Using a systematic and unbiased sequencing strategy with locus-level miRNA resolution, we profiled the first-trimester plasma miRNome and prioritized a set of 18 mature miRNAs from among 255 detected species. Set-level functional enrichment analyses based on curated and predicted miRNA–target interactions derived primarily from cellular and tissue-based studies showed annotation-based convergence on pathways related to Ca²⁺ homeostasis, glucagon–insulin regulatory circuits, and PI3K–AKT signaling. Network analysis indicated coordinated associations among these miRNAs and shared target pathways involved in insulin secretion and insulin sensitivity. Key contributors—including miR-29a-3p, miR-29c-3p, miR-146a-5p, let-7a-5p, and miR-182-5p—were linked, through in silico target annotation, to central metabolic regulators such as PTEN, PIK3R1, AKT1, AKT2, and components of Ca²⁺ signaling (ATP2A2, CALM1/3, ITPR1, RYR2). These circulating miRNAs should be interpreted primarily as biomarkers reflecting coordinated metabolic states rather than as direct causal mediators. Most identified miRNAs have not been previously reported in the context of first-trimester GDM, supporting the exploratory and hypothesis-generating nature of this circulating miRNA signature in early gestational metabolic research. Full article

Intravenous thrombolysis with recombinant tissue-type plasminogen activator (tPA) remains a keystone of acute ischemic stroke treatment but in a subset of patients is complicated by angioedema, a potentially life-threatening adverse event largely mediated by bradykinin signaling. The unpredictable and idiosyncratic nature of this reaction has long suggested an underlying genetic contribution, yet its molecular architecture has remained poorly characterized. We hypothesized that alteplase-associated angioedema represents a multigenic susceptibility phenotype, arising from the convergence of rare genetic variants across multiple interacting physiological systems rather than from a single causal variant. To explore this hypothesis, we performed clinical exome sequencing in a cohort of 11 patients who developed angioedema following alteplase administration. Rather than identifying a shared pathogenic variant, we observed distinct yet convergent patterns of genetic vulnerability, allowing patients to be grouped according to dominant, but overlapping, biological axes. These included alterations affecting bradykinin regulation (e.g., ACE, SERPING1, XPNPEP2), endothelial structure and hemostasis (e.g., VWF, COL4A1), neurovascular and calcium signaling (e.g., SCN10A, RYR1), and vascular repair or remodeling pathways (e.g., PSEN2, BRCA2). Notably, many of the identified variants were classified as Variant of Uncertain Significance (VUS) or likely benign significance in isolation. However, when considered within an integrated, pathway-based framework, these variants can be interpreted as capable of contributing cumulatively to system level fragility, a phenomenon best described as “contextual pathogenicity”. Under the acute biochemical and proteolytic stress imposed by thrombolysis, this reduced physiological reserve may allow otherwise compensated vulnerabilities to become clinically manifest. Together, these findings support a model in which severe alteplase-associated angioedema appears as an emergent property of interacting genetic networks, rather than a monogenic disorder. This systems level perspective underscores the limitations of gene centric interpretation for adverse drug reactions and highlights the potential value of pathway informed, multi-genic approaches to risk stratification. Such frameworks may ultimately contribute to safer, more personalized thrombolytic decision, while providing a conceptual foundation for future functional and translational studies. Full article