title: Microparticles engineered to highly express peroxisome proliferator-activated receptor-γ decreased inflammatory mediator production and increased adhesion of recipient monocytes.
abstract: Circulating blood microparticles are submicron vesicles released primarily by megakaryocytes and platelets that act as transcellular communicators. Inflammatory conditions exhibit elevated blood microparticle numbers compared to healthy conditions. Direct functional consequences of microparticle composition, especially internal composition, on recipient cells are poorly understood. Our objective was to evaluate if microparticle composition could impact the function of recipient cells, particularly during inflammatory provocation. We therefore engineered the composition of megakaryocyte culture-derived microparticles to generate distinct microparticle populations that were given to human monocytes to assay for influences recipient cell function. Herein, we tested the responses of monocytes exposed to either control microparticles or microparticles that contain the anti-inflammatory transcription factor, peroxisome proliferator-activated receptor-γ (PPARγ). In order to normalize relative microparticle abundance from two microparticle populations, we implemented a novel approach that utilizes a Nanodrop Spectrophotometer to assay for microparticle density rather than concentration. We found that when given to peripheral blood mononuclear cells, microparticles were preferentially internalized by CD11b+ cells, and furthermore, microparticle composition had a profound functional impact on recipient monocytes. Specifically, microparticles containing PPARγ reduced activated monocyte production of the proinflammatory cytokines interleukin-8 and monocyte chemotactic protein-1 compared to activated monocytes exposed to control microparticles. Additionally, treatment with PPARγ microparticles greatly increased monocyte cell adherence. This change in morphology occurred simultaneously with increased production of the key extracellular matrix protein, fibronectin and increased expression of the fibronectin-binding integrin, ITGA5. PPARγ microparticles also changed monocyte mRNA levels of several genes including those under PPARγ control. Overall, the delivery of PPARγ from microparticles to human monocytes influenced gene expression, decreased inflammatory mediator production and increased monocyte adherence. These results support the concept that the composition of blood microparticles has a profound impact on the function of cells with which they interact, and likely plays a role in vascular inflammation.
authors.affiliation: Department of Microbiology and Immunology, School of Medicine and Dentistry, University of Rochester, Rochester, United States of America; Department of Environmental Medicine, School of Medicine and Dentistry, University of Rochester, Rochester, United States of America.
authors.forename: Julie
authors.initials: J
authors.lastname: Sahler
authors.affiliation: Department of Environmental Medicine, School of Medicine and Dentistry, University of Rochester, Rochester, United States of America.
authors.forename: Collynn F
authors.initials: CF
authors.lastname: Woeller
authors.affiliation: Department of Microbiology and Immunology, School of Medicine and Dentistry, University of Rochester, Rochester, United States of America; Department of Environmental Medicine, School of Medicine and Dentistry, University of Rochester, Rochester, United States of America.
authors.forename: Richard P
authors.initials: RP
authors.lastname: Phipps
mesh_terms: D039481:CD11b Antigen / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion; D002460:Cell Line; D055252:Cell-Derived Microparticles / Q000737:chemistry / Q000378:metabolism*; D018932:Chemokine CCL2 / Q000096:biosynthesis / Q000378:metabolism*; D004705:Endocytosis; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D016023:Integrins / Q000235:genetics / Q000378:metabolism; D016209:Interleukin-8 / Q000096:biosynthesis / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism; D008533:Megakaryocytes / Q000737:chemistry* / Q000378:metabolism; D060847:Metabolic Engineering; D009000:Monocytes / Q000166:cytology / Q000378:metabolism; D047495:PPAR gamma / Q000235:genetics* / Q000378:metabolism; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: C111835:CCL2 protein, human; D039481:CD11b Antigen; D018932:Chemokine CCL2; D005353:Fibronectins; C509295:ITGAM protein, human; D016023:Integrins; D016209:Interleukin-8; D047495:PPAR gamma; D012333:RNA, Messenger