title: Silencing of Tumor Necrosis Factor Receptor-1 in Human Lung Microvascular Endothelial Cells Using Particle Platforms for siRNA Delivery.
abstract: Acute lung injury (ALI) and its most severe manifestation, acute respiratory distress syndrome (ARDS), is a clinical syndrome defined by acute hypoxemic respiratory failure and bilateral pulmonary infiltrates consistent with edema. In-hospital mortality is 38.5% for AL, and 41.1% for ARDS. Activation of alveolar macrophages in the donor lung causes the release of pro-inflammatory chemokines and cytokines, such as TNF-α. To determine the relevance of TNF-α in disrupting bronchial endothelial cell function, we stimulated human THP-1 macrophages with lipopolysaccharide (LPS) and used the resulting cytokine-supplemented media to disrupt normal endothelial cell functions. Endothelial tube formation was disrupted in the presence of LPS-activated THP- 1 conditioned media, with reversal of the effect occurring in the presence of 0.1µg/ml Enbrel, indicating that TNF-α was the major serum component inhibiting endothelial tube formation. To facilitate lung conditioning, we tested liposomal and porous silicon (pSi) delivery systems for their ability to selectively silence TNFR1 using siRNA technology. Of the three types of liposomes tested, only cationic liposomes had substantial endothelial uptake, with human cells taking up 10-fold more liposomes than their pig counterparts; however, non-specific cellular activation prohibited their use as immunosuppressive agents. On the other hand, pSi microparticles enabled the accumulation of large amounts of siRNA in endothelial cells compared to standard transfection with Lipofectamine(®) LTX, in the absence of non-specific activation of endothelia. Silencing of TNFR1 decreased TNF-α mediated inhibition of endothelial tube formation, as well as TNF-α-induced upregulation of ICAM-1, VCAM, and E-selection in human lung microvascular endothelial cells.
authors.forename: Litao
authors.initials: L
authors.lastname: Bai
authors.forename: Helen A
authors.initials: HA
authors.lastname: Andersson
authors.forename: Kellie I
authors.initials: KI
authors.lastname: McConnell
authors.forename: Diana L
authors.initials: DL
authors.lastname: Chan
authors.forename: Michael
authors.initials: M
authors.lastname: Hernandez
authors.forename: Javier
authors.initials: J
authors.lastname: Gonzalez
authors.forename: Xuewu
authors.initials: X
authors.lastname: Liu
authors.forename: Saverio
authors.initials: S
authors.lastname: La Francesca
authors.forename: Jason H
authors.initials: JH
authors.lastname: Sakamoto
authors.affiliation: Baylor College of Medicine, Michael E. DeBakey Department of Surgery, One Baylor Plaza, R512, Houston, TX 77030, USA. rita.serda@bcm.edu.
authors.forename: Rita E
authors.initials: RE
authors.lastname: Serda
mesh_terms: D055371:Acute Lung Injury / Q000503:physiopathology*; D000818:Animals; D002412:Cations / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D019040:E-Selectin / Q000235:genetics; D042783:Endothelial Cells / Q000378:metabolism; D020868:Gene Silencing; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D008081:Liposomes; D008264:Macrophages / Q000378:metabolism; D055806:Microvessels / Q000166:cytology / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage*; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000235:genetics*; D012128:Respiratory Distress Syndrome / Q000503:physiopathology*; D013045:Species Specificity; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D015854:Up-Regulation / Q000235:genetics; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics
chemical_list: D002412:Cations; D016207:Cytokines; D019040:E-Selectin; D008070:Lipopolysaccharides; D008081:Liposomes; D034741:RNA, Small Interfering; D047888:Receptors, Tumor Necrosis Factor, Type I; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1