Transcriptomic Analysis of Human Brain -Microvascular Endothelial Cell Driven Changes in -Vascular Pericytes

Kurmann, Lisa; Okoniewski, Michał; Dubey, Raghvendra K.

doi:10.3390/cells10071784

Cited by 9 publications

(14 citation statements)

References 92 publications

(132 reference statements)

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“…Banks and colleagues found that full-length tau and various truncated tau proteins readily crossed the BBB bidirectionally and entered the blood following injection into the brain [ 8 ], consistent with our intracranial tau injection studies. In line with these BBB findings, our studies employed a recently described indirect co-culture BBB model [ 30 ] consisting of brain endothelia and pericytes, and found the BBB transit of each tau species was greater than that observed for LyD (basolateral-to-apical). In comparing the tau species, the aggregate enriched tau showed increased BBB transcytosis compared to monomeric tau, potentially explaining the enhanced elimination of these species in the tau residence studies above.…”

Section: Discussionsupporting

confidence: 67%

Influence of traumatic brain injury on extracellular tau elimination at the blood–brain barrier

Eisenbaum

Pearson

Gratkowski

et al. 2021

Fluids Barriers CNS

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Repetitive head trauma has been associated with the accumulation of tau species in the brain. Our prior work showed brain vascular mural cells contribute to tau processing in the brain, and that these cells progressively degenerate following repetitive mild traumatic brain injury (r-mTBI). The current studies investigated the role of the cerebrovasculature in the elimination of extracellular tau from the brain, and the influence of r-mTBI on these processes. Following intracranial injection of biotin-labeled tau, the levels of exogenous labeled tau residing in the brain were elevated in a mouse model of r-mTBI at 12 months post-injury compared to r-sham mice, indicating reduced tau elimination from the brain following head trauma. This may be the result of decreased caveolin-1 mediated tau efflux at the blood–brain barrier (BBB), as the caveolin inhibitor, methyl-β-cyclodextrin, significantly reduced tau uptake in isolated cerebrovessels and significantly decreased the basolateral-to-apical transit of tau across an in vitro model of the BBB. Moreover, we found that the upstream regulator of endothelial caveolin-1, Mfsd2a, was elevated in r-mTBI cerebrovessels compared to r-sham, which coincided with a decreased expression of cerebrovascular caveolin-1 in the chronic phase following r-mTBI (> 3 months post-injury). Lastly, angiopoietin-1, a mural cell-derived protein governing endothelial Mfsd2a expression, was secreted from r-mTBI cerebrovessels to a greater extent than r-sham animals. Altogether, in the chronic phase post-injury, release of angiopoietin-1 from degenerating mural cells downregulates caveolin-1 expression in brain endothelia, resulting in decreased tau elimination across the BBB, which may describe the accumulation of tau species in the brain following head trauma.

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Section: Discussionsupporting

confidence: 67%

Influence of traumatic brain injury on extracellular tau elimination at the blood–brain barrier

Eisenbaum

Pearson

Gratkowski

et al. 2021

Fluids Barriers CNS

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“…In two recent publications [ 38 , 40 ], we have shown the modulatory effects of brain microvascular ECs and PCs on genes and BBB function. We also observed dramatic changes in interferon-associated genes and pro- as well as anti-inflammatory cytokines.…”

Section: Discussionmentioning

confidence: 99%

“…hBVPs: Human Brain Vascular Pericytes (HBVPs, ScienCell, Carlsbad, CA, USA) between the 4th and 10th passage were cultured as described previously [ 38 ]. Briefly, hBVPs were grown in flasks coated with Poly-L-Lysine (PLL; 2 µg/cm 2 ) under standard tissue culture conditions (37 °C, 5% CO 2 ) in pericyte growing media (DMEM/F12 supplemented with antibiotic-antimycotic (AA; 100 μg/mL streptomycin, 100 μg/mL penicillin and 0.025 μg/mL amphotericin B), Glutamax (1×) and 10% FBS).…”

Section: Methodsmentioning

confidence: 99%

“…hCMEC/D3: The human Cerebral Microvascular Endothelial Cell line (hCMEC/D3) [ 39 ] was kindly provided by Dr. Pierre-Olivier Couraud (Institute COCHIN, Paris, France). Cells between the 34th and 39th passage were cultured as described before [ 38 ] on rat-tail-collagen-coated (250 μg/mL in 80% EtOH) flasks under standard tissue culture conditions (37 °C, 5% CO 2 ) in complete growing media (EC basal media (EndoGRO Basal Medium supplemented with 0.2% EndoGRO-LS Supplement, 5 ng/mL rh EGF, 4 mM L-Glutamine, 0.75 U/mL Heparin Sulfate, 50 μg/mL Ascorbic Acid, 1 ng/mL bFGF, antibiotic-antimycotic (100 μg/mL streptomycin, 100 μg/mL penicillin and 0.025 μg/mL amphotericin B) supplemented with 5% FBS. Media was changed every two or three days and cells were passaged after confluency was reached.…”

Section: Methodsmentioning

confidence: 99%

“…Microarray samples were prepared as described previously [ 38 ]: PCs were seeded alone or with ECs on the opposite side of permeable PET membrane inserts of 0.4 μm pore size and six-well format (Corning Incorporated, Corning, NY, USA, Costar 3450) as described above. After 5 days in culture (2% steroid-free FCS (charcoal-stripped) in presence of hydrocortisone), cells were treated with estradiol (10 nM) or vehicle for 48 h. Cells were then trypsinized, centrifuged and lysed in 300 μL RNA lysis buffer (Zymo Research, Irvine, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

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Estradiol Inhibits Human Brain Vascular Pericyte Migration Activity: A Functional and Transcriptomic Analysis

Kurmann

Okoniewski

Dubey

2021

Cells

Self Cite

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Stroke is the third leading cause of mortality in women and it kills twice as many women as breast cancer. A key role in the pathophysiology of stroke plays the disruption of the blood–brain barrier (BBB) within the neurovascular unit. While estrogen induces vascular protective actions, its influence on stroke remains unclear. Moreover, experiments assessing its impact on endothelial cells to induce barrier integrity are non-conclusive. Since pericytes play an active role in regulating BBB integrity and function, we hypothesize that estradiol may influence BBB by regulating their activity. In this study using human brain vascular pericytes (HBVPs) we investigated the impact of estradiol on key pericyte functions known to influence BBB integrity. HBVPs expressed estrogen receptors (ER-α, ER-β and GPER) and treatment with estradiol (10 nM) inhibited basal cell migration but not proliferation. Since pericyte migration is a hallmark for BBB disruption following injury, infection and inflammation, we investigated the effects of estradiol on TNFα-induced PC migration. Importantly, estradiol prevented TNFα-induced pericyte migration and this effect was mimicked by PPT (ER-α agonist) and DPN (ER-β agonist), but not by G1 (GPR30 agonist). The modulatory effects of estradiol were abrogated by MPP and PHTPP, selective ER-α and ER-β antagonists, respectively, confirming the role of ER-α and ER-β in mediating the anti-migratory actions of estrogen. To delineate the intracellular mechanisms mediating the inhibitory actions of estradiol on PC migration, we investigated the role of AKT and MAPK activation. While estradiol consistently reduced the TNFα-induced MAPK and Akt phosphorylation, only the inhibition of MAPK, but not Akt, significantly abrogated the migratory actions of TNFα. In transendothelial electrical resistance measurements, estradiol induced barrier function (TEER) in human brain microvascular endothelial cells co-cultured with pericytes, but not in HBMECs cultured alone. Importantly, transcriptomics analysis of genes modulated by estradiol in pericytes showed downregulation of genes known to increase cell migration and upregulation of genes known to inhibit cell migration. Taken together, our findings provide the first evidence that estradiol modulates pericyte activity and thereby improves endothelial integrity.

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Secretome of brain microvascular endothelial cells promotes endothelial barrier tightness and protects against hypoxia-induced vascular leakage

Loiola,

Hachani,

Duban-Deweer

et al. 2024

Mol Med

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Cell-based therapeutic strategies have been proposed as an alternative for brain and blood vessels repair after stroke, but their clinical application is hampered by potential adverse effects. We therefore tested the hypothesis that secretome of these cells might be used instead to still focus on cell-based therapeutic strategies. We therefore characterized the composition and the effect of the secretome of brain microvascular endothelial cells (BMECs) on primary in vitro human models of angiogenesis and vascular barrier. Two different secretome batches produced in high scale (scHSP) were analysed by mass spectrometry. Human primary CD34+-derived endothelial cells (CD34+-ECs) were used as well as in vitro models of EC monolayer (CMECs) and blood–brain barrier (BBB). Cells were also exposed to oxygen–glucose deprivation (OGD) conditions and treated with scHSP during reoxygenation. Protein yield and composition of scHSP batches showed good reproducibility. scHSP increased CD34+-EC proliferation, tubulogenesis, and migration. Proteomic analysis of scHSP revealed the presence of growth factors and proteins modulating cell metabolism and inflammatory pathways. scHSP improved the integrity of CMECs, and upregulated the expression of junctional proteins. Such effects were mediated through the activation of the interferon pathway and downregulation of Wnt signalling. Furthermore, OGD altered the permeability of both CMECs and BBB, while scHSP prevented the OGD-induced vascular leakage in both models. These effects were mediated through upregulation of junctional proteins and regulation of MAPK/VEGFR2. Finally, our results highlight the possibility of using secretome from BMECs as a therapeutic alternative to promote brain angiogenesis and to protect from ischemia-induced vascular leakage.

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Transcriptomic Analysis of Human Brain -Microvascular Endothelial Cell Driven Changes in -Vascular Pericytes

Cited by 9 publications

References 92 publications

Influence of traumatic brain injury on extracellular tau elimination at the blood–brain barrier

Influence of traumatic brain injury on extracellular tau elimination at the blood–brain barrier

Estradiol Inhibits Human Brain Vascular Pericyte Migration Activity: A Functional and Transcriptomic Analysis

Secretome of brain microvascular endothelial cells promotes endothelial barrier tightness and protects against hypoxia-induced vascular leakage

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