ABSTRACT
Intercellular adhesion molecule-1 (ICAM-1) is identified as an initiator of neuroinflammatory responses that lead to neurodegeneration and cognitive and sensory-motor deficits in several pathophysiological conditions including traumatic brain injury (TBI). However, the underlying mechanisms of ICAM-1-mediated leukocyte adhesion and transmigration and its link with neuroinflammation and functional deficits following TBI remain elusive. Here, we hypothesize that blocking of ICAM-1 attenuates the transmigration of leukocytes to the brain and promotes functional recovery after TBI. The experimental TBI was induced in vivo by fluid percussion injury (25â psi) in male and female wild-type and ICAM-1-/- mice and in vitro by stretch injury (3â psi) in human brain microvascular endothelial cells (hBMVECs). We treated hBMVECs and animals with ICAM-1 CRISPR/Cas9 and conducted several biochemical analyses and demonstrated that CRISPR/Cas9-mediated ICAM-1 deletion mitigates blood-brain barrier (BBB) damage and leukocyte transmigration to the brain by attenuating the paxillin/focal adhesion kinase (FAK)-dependent Rho GTPase pathway. For analyzing functional outcomes, we used a cohort of behavioral tests that included sensorimotor functions, psychological stress analyses, and spatial memory and learning following TBI. In conclusion, this study could establish the significance of deletion or blocking of ICAM-1 in transforming into a novel preventive approach against the pathophysiology of TBI.
Subject(s)
Brain Injuries, Traumatic , Intercellular Adhesion Molecule-1 , Animals , Female , Humans , Male , Mice , Brain/metabolism , Brain Injuries, Traumatic/metabolism , CRISPR-Cas Systems , Endothelial Cells/metabolism , Focal Adhesion Protein-Tyrosine Kinases/metabolism , Leukocytes , Paxillin , rho GTP-Binding Proteins/metabolismABSTRACT
Seven EA cluster microbacteriophages were isolated from soil collected around Philadelphia, PA, using the bacterial host Microbacterium foliorum All of these phages have a highly conserved genome with regions of diversity localized to the 3' end. In phage Phriends (EA1 cluster), this region contains an orpham gene with no known function.
