Stem cell-derived extracellular vesicles (EVs) hold therapeutic potential for hypoxia-associated injury, yet the molecular mechanisms underlying their protective effects remain incompletely defined. Herein, we performed comparative proteomic profiling of extracellular vesicles (EVs) secreted by human placenta-derived perivascular stem cells (hPPSCs) and human umbilical cord mesenchymal stem cells (hUCMSCs). We found that hPPSCs-EVs are enriched in proteasome-related proteins. Enzymatic activity assays further confirmed that proteasome activity in hPPSCs-EVs was significantly higher than that in hUCMSCs-EVs. Additionally, we observed that hPPSCs-EVs were efficiently endocytosed by human umbilical vein endothelial cells (hUVECs), leading to marked downregulation of hypoxia-inducible factor 1-alpha (HIF-1α) and intracellular ubiquitinated proteins. Importantly, this HIF-1α degradation persisted even when the host ubiquitin-proteasome system was blocked, indicating that hPPSCs-EVs can function independently of the host proteasomal pathway. These results reveal that hPPSCs-EVs deliver functional proteasomes to endothelial cells, thereby compensating for impaired protein degradation and protecting cells under hypoxic stress. Collectively, our findings provide evidence for an intercellular transfer of proteolytic capacity via stem cell-derived EVs, a mechanism that preserves endothelial proteostasis and highlights the therapeutic potential of proteasome-rich EVs for hypoxia-associated diseases.

Sepsis is a life-threatening syndrome characterized by severe immune dysregulation, frequently culminating in acute respiratory distress syndrome (ARDS) and acute kidney injury (AKI). Current supportive therapies fail to reverse the underlying pathophysiological damage. However, mesenchymal stromal cells (MSCs) have emerged as a promising therapeutic frontier due to their robust immunomodulatory, anti-inflammatory, and tissue-regenerative properties. Despite compelling preclinical evidence, translating these benefits into consistent clinical efficacy remains a major challenge. This review critically examines the biological and anatomical barriers limiting the efficacy of MSCs, particularly the pulmonary first-pass effect, which restricts the systemic delivery of viable cells to distant organs such as the kidneys. To overcome these physical limitations, we highlight the recent paradigm shift toward nanoscale, cell-free therapies, specifically MSC-derived extracellular vesicles (MSC-EVs). EVs effectively bypass pulmonary sequestration and thromboembolic risks, exerting their potent therapeutic effects through the horizontal transfer of bioactive cargo, notably microRNAs, to reprogram cellular fate and restore immune homeostasis. We also discuss the critical need for rigorous clinical trial designs, scalable good manufacturing practice protocols, and the integration of a precision medicine approach. Ultimately, incorporating validated biomarkers for targeted patient stratification will be the decisive step in unlocking the full therapeutic potential of MSCs and their derivatives in critical care.

The RNA exosome plays a critical role in surveilling nuclear mRNA biogenesis and regulating co-translational mRNA decay in the cytoplasm. Unlike canonical translation, repeat-associated non-AUG (RAN) translation of a GGGGCC hexanucleotide repeat expansion (HRE) within an intron of the C9orf72 locus leads to the synthesis of neurotoxic dipeptide-repeat (DPR) proteins, contributing to the pathogenesis of frontotemporal dementia and amyotrophic lateral sclerosis (C9-ALS/FTD). However, it remains unclear whether aberrant RAN translation is monitored and regulated co-translationally or how C9orf72 HRE (C9-HRE) mRNA is degraded during this process. Here, we demonstrate that RAN translation triggers the rapid decay of C9-HRE mRNA. During this process, the RNA exosome engages the translating ribosome-C9-HRE mRNA complex to mediate RAN translation-coupled mRNA decay. Moreover, overexpression of EXOSC3, a key subunit of the RNA exosome cap, promotes RAN translation-coupled decay of C9-HRE mRNA and suppresses DPR production. In iPSC-derived neurons, a reduction in EXOSC3 levels blocks C9-HRE mRNA decay in a translation-dependent manner, further confirming its role in RAN translation surveillance. These findings highlight the essential function of the RNA exosome, particularly EXOSC3, in mitigating RAN translation-associated toxicity and preventing pathological DPR production. This work provides insights into potential therapeutic strategies for C9-ALS/FTD and may have broader implications for other disorders involving RAN translation.

Cancer stem cells (CSCs) drive therapeutic resistance, metastasis, and tumor recurrence through reversible transitions among stem-like, differentiated, epithelial, and mesenchymal states, which are sustained by interconnected epigenetic mechanisms. To our knowledge, this is the first review to integrate curcumin-mediated regulation of DNA methylation, chromatin remodeling, and non-coding RNAs within a single CSC plasticity framework and to propose the concept of an "epigenetic collapse of CSC plasticity" as a mechanistic explanation for how curcumin may weaken stemness, state switching, and adaptive treatment resistance. Evidence was critically evaluated through structured searches of PubMed/MEDLINE, Scopus, Web of Science Core Collection, Google Scholar, and citation tracking, while direct curcumin-epigenetic evidence was distinguished from independent CSC evidence and inferential mechanistic links. Curcumin has been reported to modulate DNMT1 and locus-specific DNA methylation; regulate HDACs, p300/CBP, EZH2, H3K27me3, and BMI1; and alter selected microRNA, long non-coding RNA, and circular RNA pathways, with comparatively stronger evidence involving the miR-34 family, miR-200c, miR-21, H19, and circHN1. However, current evidence is constrained by the predominance of bulk cancer-cell models, heterogeneous formulations and exposure conditions, and the scarcity of epigenetic rescue experiments combined with rigorous functional CSC assays. By unifying previously fragmented epigenetic evidence, this review advances a new evidence-weighted model in which curcumin may suppress CSC persistence not through a single molecular target, but by destabilizing the multilayer epigenetic circuitry that enables plasticity. Curcumin should therefore be regarded as a context-dependent, multilayer epigenetic modulator rather than an established CSC-eradicating therapy, and its translational relevance requires validation in prospectively defined CSC models with pharmacologically justified delivery and exposure conditions.

Duchenne muscular dystrophy (DMD) is caused by mutations in the DMD gene that abolish dystrophin expression. Prime editing enables precise genome modification without generating double-strand DNA breaks or requiring donor DNA templates. We established an in vitro prime editing workflow to generate and subsequently correct the mdx5cv mutation in mouse C2C12 myoblasts. Following optimization of engineered prime editing guide RNAs (epegRNAs) and PAM-flexible prime editors, wild-type cells were edited, clonally isolated, and genotyped. Mutation correction was then evaluated using optimized epegRNA designs. Two rounds of prime editing introduced the mdx5cv mutation into approximately 20% of alleles in C2C12 cells creating the mdx5cv C2C12 cell line. Clonal isolation yielded five homozygous mutant clones among 59 expanded colonies. Optimization studies identified an epegRNA containing a 16 nucleotide reverse transcription template and a 10 nucleotide primer binding site (RTT16/PBS10) as the most efficient design. Correction of the pathogenic allele reached approximately 26%, whereas longer PBS lengths reduced editing efficiency. In silico off-target analysis using Cas-OFFinder identified no candidate genomic loci with fewer than three mismatches for the spacer sequences used in either mutation generation or correction, suggesting a favorable predicted specificity profile. Corrected mdx5cv C2C12 myoblasts retained their capacity to differentiate into multinucleated myotubes. Representative Western blot analysis detected dystrophin protein expression in differentiated corrected mdx5cv myotubes, consistent with successful correction of the pathogenic mutation. These findings establish a robust prime editing platform for both the generation and correction of the mdx5cv mutation and provide proof of concept that precise correction of the pathogenic mutation is associated with restoration of dystrophin expression following myogenic differentiation.

Castration-resistant prostate cancer (CRPC) is characterised by persistent androgen receptor (AR)-axis activity, therapy-driven resistance, and limited durability of available systemic treatments. Tumour-tropic mesenchymal stem/stromal cells (MSCs), including adipose-derived MSCs (ADSCs), have emerged as promising vehicles for targeted gene therapeutics. This review synthesises our three experimental studies examining stem cell-delivered gene-directed enzyme prodrug therapy (GDEPT) using cytosine deaminase (CD)/5-fluorocytosine (5-FC) and secreted TRAIL in CRPC xenograft models. We performed a comparative analysis of three studies in which hTERT-immortalised human ADSCs were engineered via lentiviral vectors to deliver CD alone, secreted TRAIL alone, or CD+TRAIL in combination, and were administered by intracardiac injection into male nude mice bearing PC3 xenografts. In vitro conversion efficiency, cell viability, apoptosis markers, and in vivo tumour volume endpoints were compared across studies. All three therapeutic platforms demonstrated measurable tumour growth inhibition relative to controls. The CD+TRAIL combination achieved the greatest in vivo efficacy (tumours approximately 26% of control at day 14), compared with CD alone (approximately 71%) or TRAIL paired with irinotecan. Enzymatic conversion of 5-FC to 5-FU exceeded 93% in conditioned medium. Primary translational risks include thrombotic events associated with systemic MSC dosing, tumourigenicity and genotoxicity of hTERT-immortalised, integrating-vector-engineered cells, immunogenicity of xenogeneic CD enzyme, and systemic 5-fluorouracil leakage from flucytosine metabolism. Stem cell-delivered CD/5-FC and TRAIL constitutes a biologically rational, modular strategy for local cytotoxicity and resistance circumvention in CRPC. Successful clinical translation will require resolution of delivery-route feasibility, thrombosis risk mitigation, and a rigorous investigational new drug (IND)-enabling safety package.

Altered post-lanosterol cholesterol biosynthesis causes a heterogeneous group of rare inherited metabolic disorders, including Smith-Lemli-Opitz syndrome, desmosterolosis, lathosterolosis, and congenital hemidysplasia with ichthyosiform nevus and limb defects syndrome. These conditions are characterized by impaired cholesterol synthesis together with the accumulation of disease-specific sterol intermediates. Current experimental and clinical evidence suggests that pathogenesis reflects both cholesterol insufficiency and sterol-mediated toxicity, including oxidative stress, perturbed developmental signaling, membrane dysfunction, and impaired neurodevelopment. Experimental models have played a central role in elucidating these mechanisms and in evaluating emerging therapeutic strategies. This review provides a comprehensive overview of currently available experimental models used to investigate inherited cholesterol biosynthesis disorders, including genetically engineered animal models, patient-derived fibroblasts, immortalized and CRISPR/Cas9-edited cell lines, and induced pluripotent stem cell-based systems. Particular emphasis is placed on Smith-Lemli-Opitz syndrome, the most extensively studied disorder within this group, while recent advances in modeling desmosterolosis, lathosterolosis, and congenital hemidysplasia with ichthyosiform nevus and limb defects syndrome are also critically discussed. We compare the strengths and limitations of each experimental platform, highlighting their contributions to understanding sterol metabolism, developmental abnormalities, and cell-type-specific disease mechanisms. Finally, we discuss current challenges and future perspectives, including the development of patient-specific induced pluripotent stem cell models, genome editing approaches, and next-generation multicellular systems. Collectively, this review provides an updated framework for selecting appropriate experimental models to investigate cholesterol biosynthesis disorders and accelerate the development of mechanism-based therapeutic strategies.

Stress urinary incontinence (SUI) is a highly prevalent condition associated with pelvic floor damage, fibroblast dysfunction, and impaired extracellular matrix (ECM) remodeling. This study aimed to investigate the regenerative potential and underlying molecular mechanisms of decidua-derived mesenchymal stromal cells (DMSCs) in a rat model of vaginal distension (VD) and in human suburethral fibroblasts from SUI patients. Adult female rats were subjected to VD and treated with periurethral DMSC injections, followed by functional and transcriptomic analyses, one and six weeks after VD. In parallel, an in vitro co-culture system was used to evaluate the paracrine effects of DMSCs on SUI fibroblasts. DMSC treatment significantly increased leak point pressure (LPP) one week after VD, approaching values observed in control animals (19.8 ± 0.45 vs. 22.3 ± 2.08 mmHg, p = 0.09), while promoting transcriptional changes consistent with tissue repair. Gene expression analyses revealed transient increases in proliferative and inflammatory markers, followed by earlier normalization compared to untreated animals. In vitro, DMSCs reduced p16 expression, increased p21 and Klotho levels, and rebalanced ECM remodeling by decreasing MMP-1 and increasing MMP-2. These findings indicate that DMSCs promote a regenerative microenvironment by modulating senescence and ECM dynamics. Overall, DMSCs may represent a promising disease-modifying strategy for SUI by enhancing tissue repair and functional recovery.

The adeno-associated virus (AAV) has become the vector of choice for gene therapy and experimental gene delivery, owing to its non-pathogenic nature and ability to achieve persistent gene expression across diverse tissues. AAV has emerged as a key platform in cellular reprogramming and regenerative medicine, with applications spanning transcription factor delivery for in vivo lineage conversion and tissue repair across the CNS, heart, and musculoskeletal systems. However, significant limitations remain, particularly in the context of induced pluripotent stem cell (iPSC) engineering. We assess barriers to efficient iPSC transduction including receptor-dependent entry deficits and activation of p53-dependent DNA damage responses. Although AAV is widely described as non-integrating, evidence indicates that integration events occur in rapidly proliferating and actively reprogramming cells. Critically, we synthesize evidence that cell-type-specific promoters lose fidelity when paired with neurogenic transgene payloads, a cross-tissue problem not addressed in existing AAV reviews, and that published in vivo reprogramming efficiencies may be substantially confounded by promoter leakage in the absence of formal lineage tracing. These aspects, underrepresented in recent platform-level reviews, are specifically emphasized here as a resource for researchers designing rigorous AAV-based reprogramming and gene therapy strategies.

Obstructive sleep apnea (OSA) syndrome is characterized by repetitive nocturnal airway obstruction and is associated with intermittent hypoxia (IH). The leading cause of death among OSA patients is cardiovascular morbidity, which is greatly enhanced by IH. Despite the existence of standard treatment, cardiovascular morbidity remains unaddressed. Given the central role of IH in OSA-related cardiac damage, the present study aimed to elucidate the mechanisms underlying IH-induced cardiac injury in order to better understand and potentially improve upon current therapeutic approaches. Using human embryonic stem cell-derived cardiomyocytes (hESC-CMs) as a novel in vitro model, IH was successfully induced, and its effects on key signaling pathways were investigated. Following IH exposure, significant activation of ERK1/2, ERK5, and Erbin was demonstrated. Notably, the concurrent increase in both ERK1/2 activation and Erbin expression following IH suggests a more complex regulatory relationship between these molecules than previously appreciated. Furthermore, pathway-specific inhibition of ERK1/2 and ERK5 attenuated the IH-induced decline in beating rate, with significant restoration, following normoxic recovery. This study provides an innovative approach for in vitro investigation of OSA-associated cardiovascular morbidity and supports the search for novel pharmacological agents and molecular targets to improve the diagnosis and treatment of affected patients.

People with diabetes often have a high rate of oral implant failure and unstable long-term treatment effects. It is urgent to find the potential mechanisms and effective cure methods to address the increasing demand for dental restoration in recent years. This paper aims to find an influential factor, and a probable treatment resulting from it, to improve poor osseointegration and achieve favorable bone regeneration. To find out the influential factor of poor diabetic osseointegration, we compared marginal bone loss among patients taking different hypoglycemic drugs. To verify our pre-clinical findings, we performed in vitro experiments as well as micro-CT, fluorescence intensity, and histopathological analysis methods after establishing a diabetic rat model controlled by different systemic hypoglycemic drugs. To enhance implant osseointegration and find out the underlying mechanism, we proposed the GelMA-alogliptin hydrogel system and tested the scratch wound healing, cell adhesion, CCK-8, live-dead cell staining, osteogenesis, qPCR, and Western blot methods based on diabetic rat-derived BMSCs, HUVECs and BMMS, as well as the in vivo experiments in diabetic rats. People with diabetes under DPP4i (alogliptin) exhibit the lowest marginal bone loss. The systemic alogliptin and locally delivered peri-implant GelMA-alogliptin hydrogel treatment promoted bone regeneration around diabetic implants, enhanced diabetic-derived BMSC activity and migration through GLP1R/GSK3β/β-catenin and induced osteogenic differentiation, and promoted vascular regeneration and inflammation control. Alogliptin has positive effects on the oral implant health of people with diabetes. A locally delivered GelMA-alogliptin hydrogel system exhibits competitive ability to improve diabetic bone microenvironment and enhance implant osseointegration.

Extracellular stimuli, including hormones, growth factors and nutrients in the milieu of cells often initiate intracellular changes via signaling pathways, of which the cyclic 5', 3'-adnosine monophosphate (cAMP)-dependent protein kinase (PKA) signaling pathway is prototypical. Research in the past decades has demonstrated that PKA plays versatile roles during cell proliferation and differentiation, mainly through phosphorylating a plethora of protein substrates by its protein kinase activity. Studies using model systems including yeast, neurons and mammalian germ cells indicate that PKA functionality is regulated by not only the cell-type-specific expression of its regulatory and catalytic subunits, but also the spatiotemporal distribution of its binding proteins and secondary messengers. How PKA elicits its functional specificity in a spatiotemporal manner constitutes fundamental mechanisms that regulate development, aging and regeneration. In this review, we first summarize basic aspects that drive the functional diversity of PKA and then focus on the less studied regulatory roles of PKA during synthesis of cellular proteins, the functional units of the cell. Direct links between PKA signaling and protein synthesis machinery are yet to be fully characterized. We anticipate that research in this area, combining model systems and newly developed methodologies, will continue to deepen our understanding of animal development and the etiology of human diseases.

Gastrointestinal acute radiation syndrome (GI-ARS) is a deadly consequence of radiation exposure. We hypothesized that the peptide ghrelin is enteroprotective after radiation injury, and that ghrelin promotes intestinal stem cell regeneration via the vagus nerve. We subjected mice to 12-Gy partial body irradiation (PBI) with 5% bone marrow sparing. Some mice were vagotomized prior to PBI. We then injected the mice with human ghrelin (6 nmol/mouse) or vehicle at 24, 48, and 72 h post-irradiation, and collected blood and tissues at 96 h. PBI caused an 80% reduction in plasma citrulline, 32% shorter villi, 59% fewer crypts, a 14-fold increase in TUNEL+ cells, a 9-fold increase in intestinal permeability (FD4), and 4- to 30-fold increases in bacterial translocation (16S rRNA) to the liver and mesentery. Ghrelin significantly improved all these parameters. Remarkably, vagotomy attenuated ghrelin's protective effects by 22-58%. Mechanistically, ghrelin increased proliferating crypt cells by 2.3-fold, Lgr5+ active stem cells by 2.6-fold, Clu+ revival stem cells by 3.3-fold (immunofluorescence), and Clu mRNA by 1.6-fold compared to PBI alone, and all these effects were significantly diminished by vagotomy. Thus, ghrelin mitigates GI-ARS through vagus nerve-dependent activation of Clu+ revival stem cells and Lgr5+ stem cells, identifying a novel vagal-dependent neuroenteric pathway that regulates intestinal crypt regeneration after radiation injury.

Bone regeneration remains a significant clinical challenge, particularly for large or critical-sized defects caused by trauma, disease, or congenital abnormalities. Mesenchymal stem cells (MSCs) have emerged as a promising cell source for bone tissue engineering, with their osteogenic differentiation playing a crucial role in bone repair. Biomaterials serve as scaffolds that facilitate MSC-mediated bone regeneration by providing structural support and mimicking the extracellular matrix (ECM). This review explores recent advancements in biomaterials designed to promote MSC osteogenesis through two primary approaches: biochemical and physicomechanical stimuli. Therapeutic agent-loaded scaffolds, incorporating growth factors, small molecules, gene materials, peptides, proteins, and extracellular vesicles (EVs), have been extensively studied for their ability to enhance osteogenic differentiation. However, concerns regarding toxicity, off-target effects, and regulatory limitations have led to increasing interest in biomaterials that utilize physicomechanical cues such as stiffness, viscoelasticity, topography, porosity, and dynamic forces (shear stress, compression, vibration) as alternative or complementary strategies. Furthermore, the synergistic effects of multiple physicomechanical cues are being explored to regulate MSC behavior for promoting bone regeneration. This review discusses current challenges, emerging trends, and future directions in the development of next-generation biomaterials that integrate biochemical and physicomechanical approaches for clinical applications in bone repair and regeneration.

Cortical immature neurons (cINs) represent a unique population of prenatally generated, non-dividing neurons that maintain an immature phenotype, characterized by doublecortin (DCX) and polysialylated neural cell adhesion molecule (PSA-NCAM) expression, into adulthood. Unlike canonical adult neurogenesis involving continuous neuron generation from stem cell niches, cINs constitute a distinct form of structural plasticity termed "neurogenesis without division". This review comprehensively examines the molecular markers, morphological diversity, developmental origins, and maturation trajectories of cINs across species. We highlight the striking inverse interspecies relationship between cIN abundance and canonical adult neurogenesis, reflecting distinct biophysical and structural shifts in neural plasticity mechanisms across mammalian lineages. Furthermore, we discuss factors modulating cIN phenotype, including neurotransmitter systems, stress, sensory experience, and aging. Clinical evidence implicating cIN alterations in temporal lobe epilepsy, traumatic brain injury, and stroke is evaluated, revealing potential roles in both pathological circuit remodeling and endogenous repair. Critical gaps remain regarding the molecular programs maintaining immaturity, differentiation triggers, and the functional consequences of circuit integration. Understanding cIN biology offers new perspectives on cortical plasticity and may inform therapeutic strategies targeting endogenous cellular reserves for brain repair.

Guided bone regeneration (GBR) heavily relies on barrier membranes, with collagen being the clinical standard material. Human amniotic epithelial cells (hAECs) represent a promising, non-controversial stem cell source with substantial osteogenic and immunomodulatory potential. This study aimed to comparatively evaluate the structural characteristics of three commercial collagen membranes (Biocollagen®, Derma®, and VantyColl®) and their influence on the biological behavior, viability, and osteogenic differentiation of hAECs and hFOB 1.19 human fetal osteoblasts. The microarchitecture was assessed via scanning electron microscopy (SEM). Biological response was evaluated over 14 days, using MTT assays, calcium and phosphorus quantification, alkaline phosphatase (ALP) activity, and quantitative real-time PCR (qRT-PCR) for osteogenic markers (Runx2, Osterix, ALP, and OPN). SEM revealed a dense lamellar structure for Biocollagen®, a fibrillar and oriented architecture for Derma®, and a highly porous network for VantyColl®. Both cell types adhered to and proliferated on all membranes. Derma® provided the best long-term proliferative support for both lineages. Conversely, VantyColl® induced robust early osteoblastic differentiation, marked by exceptional upregulation of Osterix (24.93-fold) and Runx2 (2.64-fold), though it exhibited diminished long-term hAEC viability. Ultimately, collagen membrane microarchitecture dictates cell fate; dense fibrillar networks (Derma®) favor sustained growth and late matrix maturation (OPN), whereas high-porosity scaffolds (VantyColl®) amplify early osteoinductive cascades.

Maternal obesity and a Western diet high in fat and sugars are increasing worldwide and may contribute to rising neurodevelopmental and neurobehavioral disorders in offspring. Both human and animal studies link these factors to adverse outcomes. However, the cellular mechanisms driving altered early-life neurogenesis, particularly in the hippocampus, remain unclear. To assess fructose effects on neural stem cell (NSC) differentiation and neuronal morphology, hippocampal NSCs from E12.5 C57BL/6 mouse embryos were cultured and treated with fructose (8.75 or 12.5 mM) for 7 days. Neuronal and astrocyte populations, along with neuronal morphology, were analyzed by immunofluorescence, and protein expression was assessed by Western blot. Fructose treatment significantly reduced neuronal and increased astrocyte counts, leading to a decreased neuron-to-astrocyte ratio compared to controls. These findings were supported by decreased MAP2 (neuronal) and increased GFAP (astrocyte) protein expression. Furthermore, fructose also significantly reduced neurite lengths without affecting neurite number. Morphological analysis revealed decreased soma size, reduced area/perimeter, and altered soma area-to-perimeter ratios, indicating impaired structural integrity. Thus, fructose exposure shifts NSC differentiation toward an astroglial lineage while suppressing neuronal development and impairs neuronal growth and structural complexity. Future studies are necessary to determine the influence of these cellular changes on synaptic plasticity and learning, as well as memory.

Cancer stem cell (CSC) persistence drives recurrence and therapy resistance in oral squamous cell carcinoma (OSCC), but what keeps cells locked in this stem-like state is poorly understood. In this narrative review, we propose that CSC state is sustained not by any single pathway but by joint dysregulation of three interacting cell-biological systems: membrane potential (Vmem), communication between neighbouring cells through gap junctional intercellular communication (GJIC), and the Hippo-YAP pathway. We argue that these systems act together on one common point-the YAP protein, retained in the nucleus-which switches on a SOX2-centred stemness gene programme and stabilises a self-reinforcing CSC state. Drawing on evidence from cancer genomics, developmental bioelectricity, connexin biology, and OSCC-specific studies, we reconstruct how membrane depolarisation, loss of gap junction coupling, FAT1 mutation, and Hippo pathway inactivation could converge on persistent nuclear YAP, and how betel quid-the principal risk factor across South and Southeast Asia-may engage all three systems at once. Because the model holds that each input reinforces the others, it predicts that targeting several together should displace CSC state more durably than targeting any one alone. We set out the testable predictions this framework generates.

Small-molecule kinase inhibitors offer a compelling therapeutic strategy for glioblastoma, yet their clinical efficacy remains severely limited by blood-brain barrier penetration and active efflux transporter extrusion. This review evaluates current medicinal chemistry approaches and translational paradigms to overcome these drug delivery and biological constraints. A critical analysis of the literature reveals that direct structural optimization faces a multidimensional balancing act; next-generation design must prioritize macrocyclization, structural rigidification, and bioisosteric capping to lower polar surface area and evade P-glycoprotein and BCRP efflux. Furthermore, carrier-mediated prodrugs targeting the LAT1 transporter provide a viable rescue strategy for highly potent scaffolds. Reviewing recent clinical failures, such as paxalisib and osimertinib, underscores that single-node monotherapies fail due to compensatory pathway hyperactivation and clonal heterogeneity, whereas multi-targeted agents or rational dual-node combinations prevent rapid tumor adaptation. Additionally, combining kinase inhibitors with DNA damage repair inhibitors, immune checkpoint modulation, or MR-guided focused ultrasound could provide powerful synergistic networks. Finally, bridging the translational gap requires complementing conventional serum-cultured cell lines with patient-derived glioma stem cells and orthotopic xenografts to better recapitulate the cellular architecture of the disease. Ultimately, overcoming the therapeutic challenges in glioblastoma demands a fundamental pivot toward rigorous neuro-pharmacological design and multi-lineage network oncology.

The automaticity of human-induced Pluripotent Stem Cell-derived cardiomyocytes (hiPSC-CMs) is governed by coupled Ca2+ and membrane clocks, coordinated through local Ca2+ releases (LCRs) and cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling. We investigated the role of PKA in hiPSC-CM energetics by measuring its dynamics in the cytosol and mitochondria, and its crosstalk with Ca2+. We tested three hypotheses: (i) Ca2+-activated PKA signaling regulates energy balance; (ii) adenylyl cyclase activity correlates with spontaneous beating; and (iii) PKA compartmentalization is Ca2+-dependent. We also compared hiPSC-CMs with rabbit sinoatrial node cells (SANCs). The key findings are: (i) PKA inhibition (H-89), Ca2+ chelation (BAPTA), or mitochondrial Ca2+ blockade (Ru360) led to energy imbalance; (ii) H-89 induced compartmentalized PKA activity in the cytosol, mitochondrial matrix, and outer mitochondrial membrane; (iii) Ca2+ chelation with BAPTA reduced PKA activity globally; and (iv) PKA dynamics and Ca2+-dependent regulation were similar in hiPSC-CMs and rabbit SANCs. In conclusion, intracellular Ca2+-mediated PKA compartmentalization is present in hiPSC-CMs and rabbit SANCs.