Neurons enhance blood–brain barrier function via upregulating claudin-5 and VE-cadherin expression due to glial cell line-derived neurotrophic factor secretion

eLife, Journal Year: 2024, Volume and Issue: 13

Published: May 29, 2024

Blood–brain barrier (BBB) prevents neurotoxins from entering central nervous system. We aimed to establish and characterize an in vitro triple co-culture BBB model consisting of brain endothelial cells hCMEC/D3, astrocytoma U251 cells, neuroblastoma SH-SY5Y cells. Co-culture markedly enhanced claudin-5 VE-cadherin expression hCMEC/D3 accompanied by increased transendothelial electrical resistance decreased permeability. Conditioned medium (CM) (S-CM), (U-CM), (US-CM) also promoted expression. Glial cell line-derived neurotrophic factor (GDNF) levels S-CM US-CM were significantly higher than CMs U-CM. Both GDNF upregulated expression, which attenuated anti-GDNF antibody signaling inhibitors. via the PI3K/AKT/FOXO1 MAPK/ERK pathways. Meanwhile, activating PI3K/AKT/ETS1 MAPK/ERK/ETS1 signaling. The roles integrity validated using brain-specific Gdnf silencing mice. developed was successfully applied predict In conclusion, neurons enhance upregulating through secretion established may be used drugs’

Language: Английский

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25-Hydroxycholesterol attenuates tumor necrosis factor alpha-induced blood-brain barrier breakdown in vitro

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, Journal Year: 2024, Volume and Issue: 1870(8), P. 167479 - 167479

Published: Aug. 23, 2024

Intracellular cholesterol metabolism is regulated by the SREBP-2 and LXR signaling pathways. The effects of inflammation on these molecular mechanisms remain poorly studied, especially at blood-brain barrier (BBB) level. Tumor necrosis factor α (TNFα) a proinflammatory cytokine associated with BBB dysfunction. Therefore, aim our study was to investigate TNFα metabolism, focusing its underlying Using human in vitro model composed brain-like endothelial cells (hBLECs) brain pericytes (HBPs), we observed that increases permeability degrading tight junction protein CLAUDIN-5 activating stress pathways both cell types. also promotes release decreases accumulation APOE secretion. In hBLECs, expression targets (LDLR HMGCR) increased, while ABCA1 decreased. HBPs, only LDLR increased. treatment induces 25-hydroxycholesterol (25-HC) production, metabolite involved immune response intracellular metabolism. 25-HC pretreatment attenuates TNFα-induced leakage partially alleviates ABCA1, LDLR, HMGCR expression. Overall, results suggest favors efflux via an LXR/ABCA1-independent mechanism BBB, it activates pathway. Treatment reversed effect LXR/SREBP-2 Our provides novel perspectives for better understanding cerebrovascular events linked dysfunction neuroinflammatory diseases.

Language: Английский

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Identification of key regulators in pancreatic ductal adenocarcinoma using network theoretical approach

PLoS ONE, Journal Year: 2025, Volume and Issue: 20(1), P. e0313738 - e0313738

Published: Jan. 27, 2025

Pancreatic Ductal Adenocarcinoma (PDAC) is a devastating disease with poor clinical outcomes, which mainly because of delayed detection, resistance to chemotherapy, and lack specific targeted therapies. The disease’s development involves complex interactions among immunological, genetic, environmental factors, yet its molecular mechanism remains elusive. A major challenge in understanding PDAC etiology lies unraveling the genetic profiling that governs network. To address this, we examined gene expression profile compared it healthy controls, identifying differentially expressed genes (DEGs). These DEGs formed basis for constructing protein interaction network, their network topological properties were calculated. It was found self-organizes into scale-free fractal state weakly hierarchical organization. Newman Girvan’s algorithm (leading eigenvector (LEV) method) community detection enumerated four communities leading at least one motif defined by G (3,3). Our analysis revealed 33 key regulators predominantly enriched neuroactive ligand-receptor interaction, Cell adhesion molecules, Leukocyte transendothelial migration pathways; positive regulation cell proliferation, kinase B signaling biological functions; G-protein beta-subunit binding, receptor binding functions etc. Transcription Factor mi-RNA obtained. Recognizing therapeutic potential biomarker significance Key regulators, also identified approved drugs genes. However, imperative subject experimental validation establish efficacy context PDAC.

Language: Английский

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Changes in VEGFR1 and VEGFR2 expression and endothelial cell maturity in laboratory animals with a model of Alzheimer’s disease

Bulletin of Siberian Medicine, Journal Year: 2025, Volume and Issue: 23(4), P. 47 - 54

Published: Jan. 20, 2025

Aim . To evaluate the expression of VEGFR1 and VEGFR2 maturity endothelial cells in neurogenic niches model Alzheimer’s disease. Materials methods The study was carried out on 6-month-old male C57BL/6 mice. experimental group (n = 15) received 2 µl 1 mM Aβ25-35 solution CA1 hippocampal region, while control normal saline. Brain plasticity assessed at day 10, 17, 38 after surgery by passive avoidance test. VEGFR1, VEGFR2, CLDN5 immunohistochemistry Image ExFluorer imaging system. Results In group, cognitive training stimulated angiogenesis brain, which accompanied formation microvasculature with fully mature endothelium. an early pronounced increase observed 7 training, followed impaired barrier high 28 training. These changes were associated small vessels structural incompetence cells. Conclusion Angiogenesis animals disease is characterized incompetent mechanisms regulating subpopulation composition cells, stabilization layer, a decrease maturation rate newly formed microvessels time deficit manifestation. This may contribute to microcirculatory dysfunction neurogenesis as well development pathological permeability neuroinflammation. On whole, disruption animal suggests potential contribution this mechanism aberrant brain plasticity.

Language: Английский

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Tight junction proteins in glial tumors development and progression

Frontiers in Cellular Neuroscience, Journal Year: 2025, Volume and Issue: 19

Published: Feb. 3, 2025

Tight junctions form a paracellular barrier in epithelial and endothelial cells, they regulate the diffusion of fluids, molecules, penetration cells across tissue compartments. are composed group integral membrane proteins, which include claudin family, tight junction-associated Marvel protein junctional adhesion molecule proteins that anchor cytoskeleton, such as zonula occludens cingulin family. Several factors, neurotransmitters or cytokines, processes like ischemia/hypoxia, inflammation, tumorigenesis, phosphorylation/dephosphorylation, ubiquitination, palmitoylation, junction proteins. Claudins involved tumorigenesis lead to glioma formation. In gliomas, there is noticeable dysregulation claudins, occludin, occludens-1 abundance, their dislocation has been observed. The weakening intercellular cell detachment responsible for infiltration into surrounding tissues. Furthermore, permeability blood–brain barrier, formed with involvement influences development peritumoral edema – and, simultaneously, rate drug delivery glial tumor. Understanding environments brain tumors crucial predicting tumor progression feasibility chemotherapeutic delivery. This knowledge may also illuminate differences between high low-grade gliomas.

Language: Английский

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Generation of Advanced Blood–Brain Barrier Spheroids Using Human‐Induced Pluripotent Stem Cell‐Derived Brain Capillary Endothelial‐Like Cells

Advanced Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 6, 2025

Abstract Extensively studied blood–brain barrier (BBB) in‐vitro models are established on 2D cell culture inserts. However, they do not accurately represent 3D in‐vivo microenvironments due to lack of direct neurovascular unit cellular contacts. Here, the establishment and characterization a self‐assembled BBB spheroid model using human‐induced pluripotent stem (hiPSC)‐derived brain capillary endothelial‐like cells (iBCECs) in combination with primary human astrocytes (ACs) pericytes (PCs) reported. This investigation compares spheroids mono‐cultured iBCECs derived from two different hiPSC lines differentiation strategies. It is observed that properties vary depending strategy or type line applied for generation. demonstrate like tight junction ultrastructure and, comparison models, higher transcript expression specific genes. Furthermore, possess characteristic integrity, functionality, protein expression. inferred hiPSC‐derived hold strong potential as reliable future test system.

Language: Английский

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Progression of experimental autoimmune encephalomyelitis in mice and neutrophil-mediated blood-brain barrier dysfunction requires non-muscle myosin light chain kinase

Journal of Cerebral Blood Flow & Metabolism, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 7, 2025

Blood-brain barrier (BBB) dysfunction occurs in numerous central nervous system disorders. Unfortunately, a limited understanding of the mechanisms governing function hinders identification and assessment BBB-targeted therapies. Previously, we found that non-muscle myosin light chain kinase (nmMLCK) negatively regulates tight junction protein claudin-5 brain microvascular endothelial cells (BMVECs) under inflammatory conditions. Here, used complementary animal primary cell co-culture models to further investigate nmMLCK during neuroinflammation. We nmMLCK-knockout mice resisted experimental autoimmune encephalomyelitis (EAE), including paralysis, demyelination, neutrophil infiltration, BBB dysfunction. However, transiently silencing culminated fulminant disease course. In parallel, neutrophil-secreted factors triggered biphasic loss quality wild-type BMVEC monolayers, plus pronounced migration second phase. Conversely, monolayers migration. Lastly, an inverse relationship between expression BMVECs Overall, our findings support pathogenic role for EAE includes reveal contributes immune properties BMVECs, underscore harmful effects

Language: Английский

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Key Claudins at the Blood-Retina Barriers

Advances in experimental medicine and biology, Journal Year: 2025, Volume and Issue: unknown, P. 447 - 451

Published: Jan. 1, 2025

Language: Английский

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Oroxylin A Attenuates Homocysteine-Induced Blood–Brain Barrier (BBB) Dysfunction by Reducing Endothelial Permeability and Activating the CREB/Claudin-5 Signaling Pathway

ACS Chemical Neuroscience, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 25, 2025

Recent reports have indicated that elevated levels of homocysteine (Hcy) are closely linked to blood-brain barrier (BBB) dysfunction in neurological disorders. Oroxylin A (OA) is a key bioactive flavonoid has been reported regulate brain functions. However, the role OA Hcy-related BBB less reported. In this study, we aimed elucidate and molecular mechanism Hcy-mediated using both vivo vitro investigations. Our findings indicate expression tight junction (TJ) protein Claudin-5 declined, diffusion sodium fluorescein brains Hcy-challenged mice. These effects were notably rescued by administration OA. bEnd.3 microvascular endothelial cells, increased permeability, reduced trans-endothelial electrical resistance (TEER), downregulated observed. significantly reversed 25 50 μM Interestingly, treatment restored dephosphorylation CREB at Ser133 induced Hcy. addition kinase A/cAMP-response element binding (PKA/CREB) inhibitor H89 counteracted protective on inhibiting permeability promoting expression. Together, demonstrate protects against Hcy-induced maintaining integrity barriers. This effect achieved through activation CREB/Claudin-5 signaling pathway, highlighting potential therapeutic value addressing BBB-related

Language: Английский

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Mechanisms of receptor-mediated transcytosis at the blood-brain barrier

Journal of Controlled Release, Journal Year: 2025, Volume and Issue: 381, P. 113595 - 113595

Published: March 6, 2025

In receptor-mediated transcytosis (RMT) of large therapeutics across the blood-brain barrier (BBB), construct - a macromolecule or larger carrier with therapeutic payload binds protein on brain capillary endothelial cells (BCEC), internalization and release into parenchyma. The construct's into, trafficking from, but also possible entrapment within BCEC are affected by its engineered properties whose optimization has helped derive insights transport mechanisms at BCEC. Furthermore, advances in multi-omics, as well large-scale screening directed evolution campaigns have identify new targets for RMT this perspective, I raise reflect some fundamental questions one can arrive comparing BBB-targeted constructs different target proteins. These concern underlying, transcytosis-promoting factors that constructs' appears to converge on, precise role proteins RMT, through which these may mediate trafficking, tentative criteria selection Based considerations propose several scenarios strategies interfere more efficient internalization, endosomal network toward abluminal membrane, from BCEC, both smaller macromolecules carriers.

Language: Английский

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