Objective. The outcome of directed differentiation for cell-based therapies is profoundly influenced by the starting cell's intrinsic state, or cellular context. How pluripotent versus lineage-primed cells interpret the same developmental cues remains unclear. To address this, we performed a comparative transcriptomic analysis using two mouse cell lines engineered for episomal expression: a pluripotent embryonic stem cell line (PLT-ES) and a multipotent pancreatic stem cell line (PLT-PPPD). Methods. We introduced a core set of pancreatic transcription factors - PMN: Pdx1 (pancreatic and duodenal homeobox 1), MafA (MAF BZIP transcription factor A), and NeuroD1 (neurogenic differentiation 1) - into both cell types and analyzed global gene expression and insulin promoter activity. Results. The two cell types exhibited strikingly divergent fates. In lineage-primed PLT-PPPD cells, PMN expression induced a mixed pancreatic/hepatic phenotype. In stark contrast, PMN expression in pluripotent PLT-ES cells failed to initiate an endodermal program, instead robustly driving differentiation towards mesodermal lineages. Intriguingly, reporter assays revealed that the PMN factors could activate the insulin promoter in both PLT-ES and PLT-PPPD cells, regardless of the overall differentiation outcome. Conclusions. Our findings demonstrate that cellular context is a paramount determinant of cell fate. The activation of a single promoter is insufficient to orchestrate a complete differentiation program. While pancreatic stem cells exhibit plasticity within the endodermal lineage, pluripotent cells are misdirected towards entirely different germ layers if not appropriately primed. This study underscores the critical importance of using lineage-committed cells for predictable therapeutic outcomes.
Objective. Glioblastomas are the most common and highly aggressive malignant brain tumors that are difficult to treat. Hypoxia is a significant factor in glioblastoma growth. Previous studies have shown that the inhibition of ERN1 (endoplasmic reticulum to nucleus signaling 1) significantly suppresses the glioblastoma cell proliferation and modifies the hypoxic regulation. The present study aimed to investigate the impact of hypoxia on the expression of endoplasmic reticulum stress-dependent genes including EGLN1 in U87MG glioblastoma cells in relation to ERN1 inhibition to reveal a possible role of the ERN1 signaling pathway in the hypoxic regulation of these genes' expression. Methods. The control U87MG glioblastoma cells (transfected by an empty vector) and ERN1 knockdown cells with inhibited ERN1 endoribonuclease and protein kinase (dnERN1) were used. Hypoxia was induced at normoxic conditions with dimethyloxalylglycine (0.5 mM for 4 h). RNA was extracted and reverse transcribed. The expression levels of EGLN1, HSPA5, DDIT4, PA2G4, XIAP, and MTOR genes were studied by real-time qPCR and normalized to ACTB mRNA. Results. It was established that hypoxia increases the expression level of EGLN1, HSPA5, DDIT4, and XIAP in glioblastoma cells with native ERN1. However, the expression levels of two other stress-dependent genes (PA2G4 and MTOR) were reduced in these glioblastoma cells under hypoxia. Furthermore, inhibition of ERN1 enzymatic activity attenuated the effects of hypoxia on the expression of most of the genes studied in glioblastoma cells, while XIAP gene expression was dramatically increased. Conclusion. The results of this study showed that hypoxia differentially affects the expression of genes related to endoplasmic reticulum stress in glioblastoma cells and is dependent on ERN1 activity. This reflected the ERN1-mediated reprogramming of hypoxic regulation of gene expression.
Objectives. Atherosclerosis contributes to higher mortality and morbidity in people with diabetes. Preventing future complications largely depends on the early detection of imminent atherosclerosis. This study aimed to explore the relationship between the subclinical atherosclerosis measured by carotid intima-media thickness (CIMT), ankle-brachial index (ABI), and circulating levels of adipocyte fatty acid-binding protein (AFABP) in individuals with type 2 diabetes mellitus (T2DM). Methods. A total of 80 participants evenly divided into two groups: 40 newly diagnosed patients with T2DM and 40 healthy controls. Patients with T2DM had no prior history or clinical signs of macrovascular complications, such as coronary artery disease, cerebrovascular stroke or peripheral artery disease. Serum total cholesterol, triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), fasting plasma glucose (FPG), and glycated hemoglobin (HbA1c) were measured. Plasma AFABP levels were determined using a human enzyme-linked immunosorbent assay (ELISA) kit. ABI was assessed using a sphygmomanometer with a cuff that fits the limb's circumference and a portable Doppler device. Doppler ultrasound of the carotid artery was used to measure CIMT. Results. In patients with T2DM, circulating AFABP levels were significantly higher than those in healthy subjects (p=0.007). Multivariate regression analysis indicated that AFABP is the only independent factor influencing the maximum CIMT (β=0.004, 95% CI: 0.001-0.007, p=0.017). Conclusion. Plasma AFABP level could serve as a biomarker for early diagnosis of atherosclerosis in patients with T2DM.
Multiple sclerosis (MS) is a chronic immune-mediated demyelinating disease of the central nervous system characterized by inflammation, reactive gliosis, and progressive neuroaxonal damage resulting in heterogeneous clinical and histopathological manifestations. As MS often leads to disability at a young age, it represents a substantial socio-economic burden in developed countries. The etiopathogenesis of MS is multifactorial and incompletely understood, involving genetic, immunologic, and environmental factors. Recent research highlights immune responses to Epstein-Barr virus, blood-brain barrier disruption, microbiome-gut-brain axis alterations, oxidative damage, and mitochondrial dysfunction. Studying patients with newly diagnosed MS without significant comorbidities provides insight into early disease mechanisms before disability development or long-term treatment effects. This mini-review focuses on early vascular and metabolic alterations that may contribute to MS, including lipoprotein subfractions as markers of incipient atherosclerosis, endothelial dysfunction as an initiating vascular event, and autonomic nervous system imbalance during disease progression. It also addresses insulin sensitivity as a key metabolic factor alongside chronic inflammation and oxidative damage as interconnected mechanisms driving tissue injury. Metabolic changes reflecting neuronal impairment, mitochondrial dysfunction, and astroglial activation are detectable in both lesional and normal-appearing white matter in early stages. Reduced antioxidant capacity supports a role of oxidative damage in MS pathogenesis. Accelerated vascular aging, independent of traditional cardiovascular risk factors, may progress from endothelial dysfunction to structural atherosclerotic changes. Subtle alterations in lipoprotein profiles further suggest an increased risk of atherosclerosis, potentially influenced by inflammatory activity and oxidative damage, with possible sex-specific differences. Autonomic dysfunction appears to develop secondary to disease progression rather than as a primary driver of pathogenesis.
The early life experiences have an important impact on the development of the brain and behavior and early life adversities (ELA) may affect several biological systems including the hypothalamic-pituitary-adrenal (HPA) axis, neurotransmitter and immune signaling systems, and microbiota composition. Dysregulation of these systems may result in an altered stress reactivity in both early life and the adulthood periods leading to maladaptive responses to the environmental stimuli. The activation of certain neuropeptides, including oxytocin, stimulation of the HPA axis, and increased glucocorticoid levels, may also play an important role in the early adaptive processes. In terms of brain maturation, ELA can directly or indirectly elicit structural changes in neurite growth, neurogenesis, neuronal connectivities, and signaling processes, which may contribute to the production of the long-term behavioral changes associated with an increased risk of the neuropsychiatric disorders' development in later periods of the life. In this review, we summarize the effect of ELA on the HPA axis function, stress-related hormonal balance, immune responses, and the gut microbiome indicating how these changes may affect the brain function and behavior in the early stages of the life and adulthood. We also provide insight into animal studies revealing the responses of corticotropin-releasing hormone, urocortins, and corticosterone in various neural circuits in response to ELA evoked by maternal separation and limited bedding paradigms.
