Perlecan domain V is neuroprotective and proangiogenic following ischemic stroke in rodents
Boyeon Lee, … , Sarah A. Thomas, Gregory J. Bix
Boyeon Lee, … , Sarah A. Thomas, Gregory J. Bix
Published July 11, 2011
Citation Information: J Clin Invest. 2011;121(8):3005-3023. https://doi.org/10.1172/JCI46358.
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Research Article Angiogenesis

Perlecan domain V is neuroprotective and proangiogenic following ischemic stroke in rodents

Abstract

Stroke is the leading cause of long-term disability and the third leading cause of death in the United States. While most research thus far has focused on acute stroke treatment and neuroprotection, the exploitation of endogenous brain self-repair mechanisms may also yield therapeutic strategies. Here, we describe a distinct type of stroke treatment, the naturally occurring extracellular matrix fragment of perlecan, domain V, which we found had neuroprotective properties and enhanced post-stroke angiogenesis, a key component of brain repair, in rodent models of stroke. In both rat and mouse models, Western blot analysis revealed elevated levels of perlecan domain V. When systemically administered 24 hours after stroke, domain V was well tolerated, reached infarct and peri-infarct brain vasculature, and restored stroke-affected motor function to baseline pre-stroke levels in these multiple stroke models in both mice and rats. Post-stroke domain V administration increased VEGF levels via a mechanism involving brain endothelial cell α5β1 integrin, and the subsequent neuroprotective and angiogenic actions of domain V were in turn mediated via VEGFR. These results suggest that perlecan domain V represents a promising approach for stroke treatment.

Authors

Boyeon Lee, Douglas Clarke, Abraham Al Ahmad, Michael Kahle, Christi Parham, Lisa Auckland, Courtney Shaw, Mehmet Fidanboylu, Anthony Wayne Orr, Omolara Ogunshola, Andrzej Fertala, Sarah A. Thomas, Gregory J. Bix

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Figure 10

DV’s effects are mediated via the α5β1 integrin in vitro.

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DV’s effects are mediated via the α5β1 integrin in vitro. (A) Mean perce...
(A) Mean percentage of brain endothelial cells (±SD) migrating toward 3% FBS (control) or DV with or without α5β1-GST soluble protein or α5 knockdown with α5 siRNA (as normalized to no chemoattractant negative control) (*P < 0.05, **P < 0.01, n = 5 separate experiments, each condition performed in triplicate per experiment) (B) Brain endothelial cells, either untreated or treated with DV or DV plus the α5β1-specific binding peptide CRRETAWAC, on Matrigel after 12 hours. Scale bar: 10 μm (C) Quantification of proliferation of brain endothelial cells ± α5 integrin plasmid after 48 hours ± DV measured via MTS assay (n = 15, mean ± SD normalized to control proliferation arbitrarily set to 100%, **P < 0.01, n = 5 separate experiments, each condition performed in triplicate per experiment). (D) Optical biosensor traces showing the association and dissociation of DV and BSA (control) with immobilized α5β1integrin at the concentrations (in M) listed; RU, relative units. (E) VEGF ELISA data from brain endothelial cell–secreted medium treated as labeled (**P < 0.01. n = 5 separate experiments, each condition performed in triplicate per experiment). (F) Schematic for DV-induced VEGF production and release in brain microvascular endothelial cells via α5β1 integrin. Inhibitors used in this study are indicated with red lines. The various proposed effects of VEGF (i.e., stimulation of angiogenesis and neuroprotection) are also schematically illustrated.