SPARC Expression by Cerebral Microvascular Endothelial Cells in Vitro and Its Influence on Blood-brain Barrier Properties

Overview

Journal J Neuroinflammation

Publisher Biomed Central

Date 2016 Sep 2

PMID 27581191

Citations 20

Authors

Samir Alkabie

Jayasree Basivireddy

Lixin Zhou

Jane Roskams

Peter Rieckmann

Jacqueline A Quandt

Affiliations

Soon will be listed here.

Abstract

Background: SPARC (secreted protein acidic and rich in cysteine) is a nonstructural, cell-matrix modulating protein involved in angiogenesis and endothelial barrier function, yet its potential role in cerebrovascular development, inflammation, and repair in the central nervous system (CNS) remains undetermined.

Methods: This study examines SPARC expression in cultured human cerebral microvascular endothelial cells (hCMEC/D3)-an in vitro model of the blood-brain barrier (BBB)-as they transition between proliferative and barrier phenotypes and encounter pro-inflammatory stimuli. SPARC protein levels were quantified by Western blotting and immunocytochemistry and messenger RNA (mRNA) by RT-PCR.

Results: Constitutive SPARC expression by proliferating hCMEC/D3s is reduced as cells mature and establish a confluent monolayer. SPARC expression positively correlated with the proliferation marker Ki-67 suggesting a role for SPARC in cerebrovascular development. The pro-inflammatory molecules tumor necrosis factor-α (TNF-α) and endotoxin lipopolysaccharide (LPS) increased SPARC expression in cerebral endothelia. Interferon gamma (IFN-γ) abrogated SPARC induction observed with TNF-α alone. Barrier function assays show recombinant human (rh)-SPARC increased paracellular permeability and decreased transendothelial electrical resistance (TEER). This was paralleled by reduced zonula occludens-1 (ZO-1) and occludin expression in hCMEC/D3s exposed to rh-SPARC (1-10 μg/ml) compared with cells in media containing a physiological dose of SPARC.

Conclusions: Together, these findings define a role for SPARC in influencing cerebral microvascular properties and function during development and inflammation at the BBB such that it may mediate processes of CNS inflammation and repair.

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References

Yunker C, Golembieski W, Lemke N, Schultz C, Cazacu S, Brodie C . SPARC-induced increase in glioma matrix and decrease in vascularity are associated with reduced VEGF expression and secretion. Int J Cancer. 2008; 122(12):2735-43. PMC: 3644882. DOI: 10.1002/ijc.23450. View

Forster C, Burek M, Romero I, Weksler B, Couraud P, Drenckhahn D . Differential effects of hydrocortisone and TNFalpha on tight junction proteins in an in vitro model of the human blood-brain barrier. J Physiol. 2008; 586(7):1937-49. PMC: 2375735. DOI: 10.1113/jphysiol.2007.146852. View

Chlenski A, Guerrero L, Peddinti R, Spitz J, Leonhardt P, Yang Q . Anti-angiogenic SPARC peptides inhibit progression of neuroblastoma tumors. Mol Cancer. 2010; 9:138. PMC: 2895596. DOI: 10.1186/1476-4598-9-138. View

Minagar A, Alexander J . Blood-brain barrier disruption in multiple sclerosis. Mult Scler. 2003; 9(6):540-9. DOI: 10.1191/1352458503ms965oa. View

Mendis D, Brown I . Expression of the gene encoding the extracellular matrix glycoprotein SPARC in the developing and adult mouse brain. Brain Res Mol Brain Res. 1994; 24(1-4):11-9. DOI: 10.1016/0169-328x(94)90112-0. View

Said N, Elmarakby A, Imig J, Fulton D, Motamed K . SPARC ameliorates ovarian cancer-associated inflammation. Neoplasia. 2008; 10(10):1092-104. PMC: 2546586. DOI: 10.1593/neo.08672. View

Charest A, Pepin A, Shetty R, Cote C, Voisine P, Dagenais F . Distribution of SPARC during neovascularisation of degenerative aortic stenosis. Heart. 2006; 92(12):1844-9. PMC: 1861285. DOI: 10.1136/hrt.2005.086595. View

Malaval L, Fournier B, Delmas P . Radioimmunoassay for osteonectin. Concentrations in bone, nonmineralized tissues, and blood. J Bone Miner Res. 1987; 2(5):457-65. DOI: 10.1002/jbmr.5650020514. View

Au E, Richter M, Vincent A, Tetzlaff W, Aebersold R, Sage E . SPARC from olfactory ensheathing cells stimulates Schwann cells to promote neurite outgrowth and enhances spinal cord repair. J Neurosci. 2007; 27(27):7208-21. PMC: 6794587. DOI: 10.1523/JNEUROSCI.0509-07.2007. View

10.

Argaw A, Gurfein B, Zhang Y, Zameer A, John G . VEGF-mediated disruption of endothelial CLN-5 promotes blood-brain barrier breakdown. Proc Natl Acad Sci U S A. 2009; 106(6):1977-82. PMC: 2644149. DOI: 10.1073/pnas.0808698106. View

11.

Lane T, Iruela-Arispe M, Johnson R, Sage E . SPARC is a source of copper-binding peptides that stimulate angiogenesis. J Cell Biol. 1994; 125(4):929-43. PMC: 2120067. DOI: 10.1083/jcb.125.4.929. View

12.

Iruela-Arispe M, Lane T, Redmond D, Reilly M, Bolender R, Kavanagh T . Expression of SPARC during development of the chicken chorioallantoic membrane: evidence for regulated proteolysis in vivo. Mol Biol Cell. 1995; 6(3):327-43. PMC: 301191. DOI: 10.1091/mbc.6.3.327. View

13.

Goldblum S, Ding X, Funk S, Sage E . SPARC (secreted protein acidic and rich in cysteine) regulates endothelial cell shape and barrier function. Proc Natl Acad Sci U S A. 1994; 91(8):3448-52. PMC: 43594. DOI: 10.1073/pnas.91.8.3448. View

14.

Iruela-Arispe M, Diglio C, Sage E . Modulation of extracellular matrix proteins by endothelial cells undergoing angiogenesis in vitro. Arterioscler Thromb. 1991; 11(4):805-15. DOI: 10.1161/01.atv.11.4.805. View

15.

Kato Y, Lewalle J, Baba Y, Tsukuda M, Sakai N, Baba M . Induction of SPARC by VEGF in human vascular endothelial cells. Biochem Biophys Res Commun. 2001; 287(2):422-6. DOI: 10.1006/bbrc.2001.5622. View

16.

Poller B, Gutmann H, Krahenbuhl S, Weksler B, Romero I, Couraud P . The human brain endothelial cell line hCMEC/D3 as a human blood-brain barrier model for drug transport studies. J Neurochem. 2008; 107(5):1358-68. DOI: 10.1111/j.1471-4159.2008.05730.x. View

17.

Bennett J, Basivireddy J, Kollar A, Biron K, Reickmann P, Jefferies W . Blood-brain barrier disruption and enhanced vascular permeability in the multiple sclerosis model EAE. J Neuroimmunol. 2010; 229(1-2):180-91. DOI: 10.1016/j.jneuroim.2010.08.011. View

18.

Porter P, Sage E, Lane T, Funk S, Gown A . Distribution of SPARC in normal and neoplastic human tissue. J Histochem Cytochem. 1995; 43(8):791-800. DOI: 10.1177/43.8.7622842. View

19.

Cucullo L, Couraud P, Weksler B, Romero I, Hossain M, Rapp E . Immortalized human brain endothelial cells and flow-based vascular modeling: a marriage of convenience for rational neurovascular studies. J Cereb Blood Flow Metab. 2007; 28(2):312-28. DOI: 10.1038/sj.jcbfm.9600525. View

20.

Abbott N, Ronnback L, Hansson E . Astrocyte-endothelial interactions at the blood-brain barrier. Nat Rev Neurosci. 2005; 7(1):41-53. DOI: 10.1038/nrn1824. View