title: Moss-Produced, Glycosylation-Optimized Human Factor H for Therapeutic Application in Complement Disorders.
abstract: Genetic defects in complement regulatory proteins can lead to severe renal diseases, including atypical hemolytic uremic syndrome and C3 glomerulopathies, and age-related macular degeneration. The majority of the mutations found in patients with these diseases affect the glycoprotein complement factor H, the main regulator of the alternative pathway of complement activation. Therapeutic options are limited, and novel treatments, specifically those targeting alternative pathway activation, are highly desirable. Substitution with biologically active factor H could potentially treat a variety of diseases that involve increased alternative pathway activation, but no therapeutic factor H is commercially available. We recently reported the expression of full-length recombinant factor H in moss (Physcomitrella patens). Here, we present the production of an improved moss-derived recombinant human factor H devoid of potentially immunogenic plant-specific sugar residues on proteinN-glycans, yielding approximately 1 mg purified moss-derived human factor H per liter of initialP. patensculture after a multistep purification process. This glycosylation-optimized factor H showed fullin vitrocomplement regulatory activity similar to that of plasma-derived factor H and efficiently blocked LPS-induced alternative pathway activation and hemolysis induced by sera from patients with atypical hemolytic uremic syndrome. Furthermore, injection of moss-derived factor H reduced C3 deposition and increased serum C3 levels in a murine model of C3 glomerulopathy. Thus, we consider moss-produced recombinant human factor H a promising pharmaceutical product for therapeutic intervention in patients suffering from complement dysregulation.
authors.affiliation: Department of Pediatrics and Adolescent Medicine, Faculty of Medicine, University of Freiburg Medical Center, Freiburg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Michelfelder
authors.affiliation: Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany.
authors.forename: Juliana
authors.initials: J
authors.lastname: Parsons
authors.affiliation: Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany.
authors.forename: Lennard L
authors.initials: LL
authors.lastname: Bohlender
authors.affiliation: Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany.
authors.forename: Sebastian N W
authors.initials: SNW
authors.lastname: Hoernstein
authors.affiliation: Greenovation Biotech GmbH, Freiburg, Germany.
authors.forename: Holger
authors.initials: H
authors.lastname: Niederkrüger
authors.affiliation: Greenovation Biotech GmbH, Freiburg, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Busch
authors.affiliation: Greenovation Biotech GmbH, Freiburg, Germany.
authors.forename: Nicola
authors.initials: N
authors.lastname: Krieghoff
authors.affiliation: Greenovation Biotech GmbH, Freiburg, Germany.
authors.forename: Jonas
authors.initials: J
authors.lastname: Koch
authors.affiliation: Greenovation Biotech GmbH, Freiburg, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Fode
authors.affiliation: Greenovation Biotech GmbH, Freiburg, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Schaaf
authors.affiliation: Greenovation Biotech GmbH, Freiburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Frischmuth
authors.affiliation: Department of Pediatrics and Adolescent Medicine, Faculty of Medicine, University of Freiburg Medical Center, Freiburg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Pohl
authors.affiliation: Leibniz Institute for Natural Product Research and Infection Biology, Friedrich Schiller University, Jena, Germany.
authors.forename: Peter F
authors.initials: PF
authors.lastname: Zipfel
authors.affiliation: Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany.|BIOSS Centre for Biological Signalling Studies, University of Freiburg, Freiburg, Germany; and.|FRIAS Freiburg Institute for Advanced Studies, University of Freiburg, Freiburg, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Reski
authors.affiliation: Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany; karsten.haeffner@uniklinik-freiburg.de eva.decker@biologie.uni-freiburg.de.
authors.forename: Eva L
authors.initials: EL
authors.lastname: Decker
authors.affiliation: Department of Pediatrics and Adolescent Medicine, Faculty of Medicine, University of Freiburg Medical Center, Freiburg, Germany; karsten.haeffner@uniklinik-freiburg.de eva.decker@biologie.uni-freiburg.de.
authors.forename: Karsten
authors.initials: K
authors.lastname: Häffner
mesh_terms: D000818:Animals; D065766:Atypical Hemolytic Uremic Syndrome / Q000188:drug therapy*; D019068:Bryopsida / Q000378:metabolism; D017242:Complement Factor H / Q000096:biosynthesis / Q000378:metabolism / Q000627:therapeutic use; D003165:Complement System Proteins; D006031:Glycosylation; D006801:Humans; D007154:Immune System Diseases / Q000188:drug therapy*; D051379:Mice
chemical_list: C493495:CFH protein, human; D017242:Complement Factor H; D003165:Complement System Proteins
keywords: C3 glomerulopathy; FHmoss; atypical hemolytic uremic syndrome; chronic kidney disease; complement; pediatric nephrology