Genetic Ablation of Extra Domain A of Fibronectin in Hypercholesterolemic Mice Improves Stroke Outcome by Reducing Thrombo-Inflammation.
BACKGROUND
The fibronectin-splicing variant containing extra domain A (Fn-EDA) is present in negligible amounts in the plasma of healthy humans but markedly elevated in patients with comorbid conditions, including diabetes mellitus and hypercholesterolemia, which are risk factors for stroke. It remains unknown, however, whether Fn-EDA worsens stroke outcomes in such conditions. We determined the role of Fn-EDA in stroke outcome in a model of hypercholesterolemia, the apolipoprotein E-deficient (Apoe(-/-)) mouse.


RESULTS
In a transient cerebral ischemia/reperfusion injury model, Apoe(-/-) mice expressing fibronectin deficient in EDA (Fn-EDA(-/-)Apoe(-/-) mice) exhibited smaller infarcts and improved neurological outcomes at days 1 and 8 (P<0.05 versus Apoe(-/-) mice). Concomitantly, intracerebral thrombosis [assessed by fibrin(ogen) deposition] and postischemic inflammation (phospho-nuclear factor-κB p65, phospho-IκB kinase α/β, interleukin 1β, and tumor necrosis factor-α) within lesions of Fn-EDA(-/-)Apoe(-/-) mice were markedly decreased (P<0.05 versus Apoe(-/-) mice). In an FeCl3 injury-induced carotid artery thrombosis model, thrombus growth rate and the time to occlusion were prolonged in Fn-EDA(-/-)Apoe(-/-) mice (P<0.05 versus Apoe(-/-) mice). Genetic ablation of TLR4 improved stroke outcome in Apoe(-/-) mice (P<0.05) but had no effect on stroke outcome in Fn-EDA(-/-)Apoe(-/-) mice. Bone marrow transplantation experiments revealed that nonhematopoietic cell-derived Fn-EDA exacerbates stroke through Toll-like receptor-4 expressed on hematopoietic cells. Infusion of a specific inhibitor of Fn-EDA into Apoe(-/-) mouse 15 minutes after reperfusion significantly improved stroke outcome.


CONCLUSIONS
Hypercholesterolemic mice deficient in Fn-EDA exhibit reduced cerebral thrombosis and less inflammatory response after ischemia/reperfusion injury. These findings suggest that targeting Fn-EDA could be an effective therapeutic strategy in stroke associated with hypercholesterolemia.
From the Department of Internal Medicine, University of Iowa, Iowa City.
Nirav
N
Dhanesha
From the Department of Internal Medicine, University of Iowa, Iowa City.
Ajmal
A
Ahmad
From the Department of Internal Medicine, University of Iowa, Iowa City.
Prem
P
Prakash
From the Department of Internal Medicine, University of Iowa, Iowa City.
Prakash
P
Doddapattar
From the Department of Internal Medicine, University of Iowa, Iowa City.
Steven R
SR
Lentz
From the Department of Internal Medicine, University of Iowa, Iowa City. anil-chauhan@uiowa.edu.
Anil K
AK
Chauhan
D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D005353:Fibronectins / Q000172:deficiency / Q000235:genetics*; D017353:Gene Deletion; D006937:Hypercholesterolemia / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D020521:Stroke / Q000235:genetics* / Q000473:pathology / Q000517:prevention & control; D016896:Treatment Outcome
D005353:Fibronectins; C572454:extra domain A fibronectin, mouse
fibronectins; hypercholesterolemia; inflammation; stroke; thrombosis