title: A comprehensive model of hyaluronan turnover in the mouse.
abstract: The metabolism of hyaluronan (HA), especially its catabolism, is still far from being elucidated. Although several studies suggest that HA is degraded locally in tissues and through the lymphatic or circulatory systems, much needs to be learned about the enzymes, receptors and cell types that support this dynamic process. In the current work, the clearance of exogenously administered HA was examined in a C57BL/6 mouse model. Hyaluronidase-sensitive fluorescein-labeled 1.2MDa hyaluronan (flHA) was administered either intravenously (i.v.) or subcutaneously (s.c.) into wild type C57BL/6 mice. Plasma was sampled for pharmacokinetic analysis and tissues were harvested for histological examination of the cell types responsible for uptake using immunofluorescent localization and for size exclusion chromatography analysis. We observed that flHA could be degraded locally in the skin or be taken up by sinusoidal cells in lymph nodes, liver and spleen. I.v. administration of flHA revealed non-linear Michaelis-Menten pharmacokinetics compatible with a saturable, receptor-mediated clearance system (K(m)=11.6μg/ml±46.0%, V(max)=1.69μg/ml/min±59.7%). Through a combination of immunofluorescence microscopy, pharmacokinetic, and chromatographic analyses of labeled substrate in vivo, our results shed additional light on the mechanisms by which HA is catabolized in mammals, and serve as a basis for future studies.
authors.affiliation: Halozyme Therapeutics Inc., San Diego, CA, USA. ljadin@halozyme.com
authors.forename: Laurence
authors.initials: L
authors.lastname: Jadin
authors.forename: Louis H
authors.initials: LH
authors.lastname: Bookbinder
authors.forename: Gregory I
authors.initials: GI
authors.lastname: Frost
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D019793:Fluorescein / Q000378:metabolism; D006820:Hyaluronic Acid / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism; D007275:Injections, Intravenous; D007279:Injections, Subcutaneous; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008198:Lymph Nodes / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D023421:Models, Animal; D008970:Molecular Weight; D012867:Skin / Q000378:metabolism*; D013154:Spleen / Q000166:cytology / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase; D019793:Fluorescein