Transport of the Proinflammatory Chemokines C-C Motif Chemokine Ligand 2 (MCP-1) and C-C Motif Chemokine Ligand 5 (RANTES) across the Intact Mouse Blood-Brain Barrier Is Inhibited by Heparin and Eprodisate and Increased with Systemic Inflammation

Quaranta, Daniel V; Weaver, Riley R; Baumann, Kristen K.; Fujimoto, Takashi; Williams, Lindsey; Kim, Hyung Chan; Logsdon, Aric F.; Omer, Mohamed; Reed, May J.; Banks, William A.; Erickson, Michelle A.

doi:10.1124/jpet.122.001380

Cited by 18 publications

(17 citation statements)

References 82 publications

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“…Basement membrane HSPGs such as agrin have also been shown to contribute to BBB maintenance and integrity (47), however we note that neither GalNaz nor HSases applied to the luminal side of iBECs reduced the barrier properties in our studies. How ow-dependent maturation of the iBEC glycocalyx impacts BBB integrity and transport of CCL2 and other substances that use HS to cross the BBB is presently unclear, although we note that CCL2 transport follows similar mechanisms in vitro as it does in vivo under ow conditions (9).…”

Section: Discussionmentioning

confidence: 77%

“…HS is a major component of the endothelial glycocalyx whose functions at the BBB are newly realized and underexplored (46,57). HS is known to be involved in the binding and activity of proteins such as growth factors and chemokines (44,58), but only recently has been identi ed as a mediator of blood-to-brain transcytosis (9,24). Our work highlights HS as an important component of some BBB transporter systems, and demonstrates a novel use of iBEC models to study the involvement of HS in the transport of substances across the intact BBB.…”

Section: Discussionmentioning

confidence: 99%

“…Blood-to-brain transport of chemokines, a subset of structurally related chemotactic cytokines, has also been demonstrated in rodents using in vitro and in vivo models (8,9). Chemokines are best known for their role in leukocyte tra cking (10), but they have also been implicated in aspects of CNS dysfunction that may be leukocyte-independent.…”

Section: Introductionmentioning

confidence: 99%

“…Chemokine receptors such as CCR2 and the duffy antigen receptor for chemokines (DARC) were shown to be particularly important transport mediators of CCL2 from the brainto-blood across the BBB (14,15). Current evidence supports that binding to HS, a glycosaminoglycan that is abundant in the glycocalyx of endothelial cells (16), contributes to the blood-to-brain transport of both CCL2 and CCL5 across the intact mouse BBB (9). Further, HS and other extracellular matrix components do not mediate binding of CCL2 to the abluminal surface of the BBB (17), suggesting that brain-to-blood and blood-to-brain transport of CCL2 and perhaps other chemokines as well are regulated by distinct mechanisms.…”

Section: Introductionmentioning

confidence: 99%

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Heparan sulfate-dependent transport of CCL2 across an in vitro model of the human blood-brain barrier

Williams

Fujimoto

Weaver

et al. 2023

Preprint

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Background: Transport of immune-active substances across the blood-brain barrier (BBB) is an important mechanism of neuroimmune regulation. CCL2 is among the chemokines known to cross the intact BBB in the blood-to-brain direction and is supported to do so in mice through interactions with heparan sulfate (HS)-containing components of the endothelial glycocalyx. The goal of this study was to characterize blood-to-brain transport mechanisms of human CCL2 in a human induced pluripotent stem-cell (iPSC)- derived in vitro model of the BBB. Methods: Human brain endothelial-like cells (iBECs) were differentiated using established methods and then changed to heparin-free medium. All experiments were conducted 9 days after seeding differentiated iBECs on permeable culture inserts or tissue culture plates. Human recombinant CCL2 and bovine serum albumin (Alb) as a leakage tracer was labeled with 125I and 131I, respectively, and their flux across the monolayer was quantified by calculating the permeability-surface area coefficient. Transport of 125I-CCL2 and 131I-Alb was evaluated at baseline, in the presence of a CCR2 inhibitor and heparin, following treatment with heparinases, and following treatment with the heparan sulfate synthesis inhibitor GalNaz to evaluate HS-dependent mechanisms of transport. We further determined the mechanism of 125I-CCL2 transcytosis using inhibitors of clathrin, caveolae, and dynamin. Results: We found that iBECs have a functional blood-to-brain transport system for CCL2. Similar to our previous findings in mice, heparin inhibited CCL2 transport whereas the CCR2 inhibitor did not. We further showed that both heparinase treatment and treatment with GalNaz inhibited CCL2 transport across the BBB, supporting the requirement for HS in CCL2 transport. CCL2 transcytosis was clathrin-independent and caveolae and dynamin-dependent. Conclusions: Our findings support that human CCL2 is transported across the human BBB in vitro by a mechanism that was HS-dependent, caveolae and dynamin-dependent, and clathrin-independent. Our findings underscore the utility of iBECs for the study of mechanisms of heparan sulfate/glycocalyx interactions in the transport of substances across the BBB.

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Section: Discussionmentioning

confidence: 77%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Heparan sulfate-dependent transport of CCL2 across an in vitro model of the human blood-brain barrier

Williams

Fujimoto

Weaver

et al. 2023

Preprint

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“…Such changes are readily observed when LPS is applied to in vitro BBB models using immunofluorescent microscopy techniques [ 9 ]. However, in vivo, a relatively high LPS dose is needed to cause widespread BBB disruption [ 9 , 23 ], whereas lower LPS doses can cause changes in transporters in the absence of detectable BBB disruption [ 45 ]. Additionally, it has been shown in another neuroinflammatory model of experimental autoimmune encephalomyelitis that tight junctions remain morphologically intact despite neuroinflammation, BBB disruption, and leukocyte trafficking [ 46 ].…”

Section: Discussionmentioning

confidence: 99%

Ultrastructural Remodeling of the Blood–Brain Barrier and Neurovascular Unit by Lipopolysaccharide-Induced Neuroinflammation

Erickson

Shulyatnikova

Banks

et al. 2023

IJMS

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The blood–brain barrier (BBB) is an interface primarily comprised of brain endothelial cells (BECs), separating the central nervous system (CNS) from the systemic circulation while carefully regulating the transport of molecules and inflammatory cells, and maintaining the required steady-state environment. Inflammation modulates many BBB functions, but the ultrastructural cytoarchitectural changes of the BBB with inflammation are understudied. Inflammation was induced in male 8–10-week-old CD-1 mice with intraperitoneal lipopolysaccharide (LPS), using a regimen (3 mg/kg at 0, 6, and 24 h) that caused robust BBB disruption but had minimal lethality at the study timepoint of 28 h. Perfusion-fixed brains were collected and the frontal cortical layer III regions were analyzed using a transmission electron microscopy (TEM). The LPS-treated mice had pronounced ultrastructural remodeling changes in BECs that included plasma membrane ruffling, increased numbers of extracellular microvesicles, small exosome formation, aberrant BEC mitochondria, increased BEC transcytosis, while tight junctions appeared to be unaltered. Aberrant pericytes were contracted with rounded nuclei and a loss of their elongated cytoplasmic processes. Surveilling microglial cells were attracted to the neurovascular unit (NVU) of BECs, and astrocyte detachment and separation were associated with the formation of a perivascular space and pericapillary edema. The LPS treatment resulted in numerous ultrastructural aberrant remodeling changes to the neurovascular unit’s BECs, microglia, pericytes, and astrocytes. In summary, a disturbance of the NVU morphology is a consequence of LPS treatment.

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The Blood–Brain Barriers

Banks,

Erickson

2024

Neuroimmune Pharmacology and Therapeutics

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Transport of the Proinflammatory Chemokines C-C Motif Chemokine Ligand 2 (MCP-1) and C-C Motif Chemokine Ligand 5 (RANTES) across the Intact Mouse Blood-Brain Barrier Is Inhibited by Heparin and Eprodisate and Increased with Systemic Inflammation

Abstract: Title: Transport of the pro-inflammatory chemokines CCL2 (MCP-1) and CCL5 (RANTES) across the intact mouse blood-brain barrier is inhibited by heparin and eprodisate and increased with systemic inflammation.

Cited by 18 publications

References 82 publications

Heparan sulfate-dependent transport of CCL2 across an in vitro model of the human blood-brain barrier

Heparan sulfate-dependent transport of CCL2 across an in vitro model of the human blood-brain barrier

Ultrastructural Remodeling of the Blood–Brain Barrier and Neurovascular Unit by Lipopolysaccharide-Induced Neuroinflammation

The Blood–Brain Barriers

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