Expression data from adult mice pulmonary endothelial cells

Sepsis patients are at increased risk for hospital-acquired pulmonary infections, potentially due to post-septic immunosuppression known as the compensatory anti-inflammatory response syndrome (CARS). CARS has been attributed to leukocyte dysfunction, with an unclear role for endothelial cells. The pulmonary circulation is lined by an endothelial glycocalyx, a heparan sulfate-rich layer essential to pulmonary homeostasis. Heparan sulfate degradation occurs early in sepsis, leading to lung injury. Endothelial synthesis of new heparan sulfates subsequently allows for glycocalyx reconstitution and endothelial recovery. We hypothesized that remodeling of the reconstituted endothelial glycocalyx, mediated by alterations in the endothelial machinery responsible for heparan sulfate synthesis, contributes to CARS. Our experimental animal model of CARS recapitulated post-septic immunosuppression, coincidentally with structural remodeling of endothelial glycocalyx heparan sulfate. We used microarray to identify which heparan sulfate modifying enzyme is responsible for the remodeling of post-septic reconstituted glycocalyx, characterized with enrichment of heparan sulfate disaccharides sulfated at the 6-O position of glucosamine. Experimental sepsis was induced in adult male mice with cecal ligation and puncture. Sham surgery was performed similarly, albeit without ligation and puncture of the cecum. The whole lungs were collected from each mouse at 48 hours after the induction of sepsis, immediately prior to the completion of heparan sulfate reconstitution. Pulmonary endothelial cells were isolated with FAC sorting (CD31+/CD144+), and total RNA was extracted and hybridized on Affymetrix microarrays.