title: Transient Expression of Functional Glucocerebrosidase for Treatment of Gaucher's Disease in the Goat Mammary Gland.
abstract: Gaucher disease (GD) is an orphan disease characterized by the lack or incapacity of glucocerebrosidase (hGCase) to properly process glucosylceramide, resulting in its accumulation in vital structures of the human body. Enzyme replacement therapy supplies hGCase to GD patients with a high-cost recombinant enzyme produced in vitro in mammalian or plant cell culture. In this study, we produced hGCase through the direct injection of recombinant adenovirus in the mammary gland of a non-transgenic goat. The enzyme was secreted in the milk during six days at a level up to 111.1 ± 8.1 mg/L, as identified by mass spectrometry, showing high in vitro activity. The milk-produced hGCase presented a mass correspondent to the intermediary high-mannose glycosylated protein, which could facilitate its delivery to macrophages through the macrophage mannose receptor. Further studies are underway to determine the in vivo delivery capacity of milk-hGCase, but results from this study paves the way toward the generation of transgenic goats constitutively expressing hGCase in the milk.
authors.affiliation: Molecular and Development Biology Laboratory, Experimental Biology Center, University of Fortaleza (UNIFOR), Fortaleza, CE, Brazil.
authors.forename: Kaio Cesar Simiano
authors.initials: KC
authors.lastname: Tavares
authors.affiliation: Quatro G Pesquisa & Desenvolvimento Ltda., Tecnopuc, Porto Alegre, RS, Brazil.
authors.forename: Ana Christina de Oliveira
authors.initials: AC
authors.lastname: Dias
authors.affiliation: Molecular and Development Biology Laboratory, Experimental Biology Center, University of Fortaleza (UNIFOR), Fortaleza, CE, Brazil.
authors.forename: Cícera Regina
authors.initials: CR
authors.lastname: Lazzarotto
authors.affiliation: Molecular and Development Biology Laboratory, Experimental Biology Center, University of Fortaleza (UNIFOR), Fortaleza, CE, Brazil.
authors.forename: Saul
authors.initials: S
authors.lastname: Gaudencio Neto
authors.affiliation: Molecular and Development Biology Laboratory, Experimental Biology Center, University of Fortaleza (UNIFOR), Fortaleza, CE, Brazil.
authors.forename: Igor
authors.initials: I
authors.lastname: de Sá Carneiro
authors.affiliation: Molecular and Development Biology Laboratory, Experimental Biology Center, University of Fortaleza (UNIFOR), Fortaleza, CE, Brazil.
authors.forename: Felipe Ledur
authors.initials: FL
authors.lastname: Ongaratto
authors.affiliation: Quatro G Pesquisa & Desenvolvimento Ltda., Tecnopuc, Porto Alegre, RS, Brazil.
authors.forename: Antônio Frederico Michel
authors.initials: AF
authors.lastname: Pinto
authors.affiliation: Molecular and Development Biology Laboratory, Experimental Biology Center, University of Fortaleza (UNIFOR), Fortaleza, CE, Brazil.
authors.forename: Luís Henrique
authors.initials: LH
authors.lastname: de Aguiar
authors.affiliation: Molecular and Development Biology Laboratory, Experimental Biology Center, University of Fortaleza (UNIFOR), Fortaleza, CE, Brazil.
authors.forename: Carlos Enrique Mendez
authors.initials: CE
authors.lastname: Calderón
authors.affiliation: Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción (Udec), Concepción, Chile.
authors.forename: Jorge Roberto
authors.initials: JR
authors.lastname: Toledo
authors.affiliation: Department of Physiopathology, Faculty of Veterinary Sciences, Universidad de Concepción (Udec), Chillán, Chile.
authors.forename: Fidel Ovidio
authors.initials: FO
authors.lastname: Castro
authors.affiliation: Quatro G Pesquisa & Desenvolvimento Ltda., Tecnopuc, Porto Alegre, RS, Brazil.
authors.forename: Diogenes Santiago
authors.initials: DS
authors.lastname: Santos
authors.affiliation: Quatro G Pesquisa & Desenvolvimento Ltda., Tecnopuc, Porto Alegre, RS, Brazil.
authors.forename: Jocelei Maria
authors.initials: JM
authors.lastname: Chies
authors.affiliation: Molecular and Development Biology Laboratory, Experimental Biology Center, University of Fortaleza (UNIFOR), Fortaleza, CE, Brazil.|School of Veterinary Medicine, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Bertolini
authors.affiliation: Molecular and Development Biology Laboratory, Experimental Biology Center, University of Fortaleza (UNIFOR), Fortaleza, CE, Brazil. lrellybertolini@gmail.com.|Biotechnology and Genetic Engineering Lab, School of Pharmacy, Pontifical Catholic University of Rio Grande do Sul (PUCRS), Porto Alegre, RS, Brazil. lrellybertolini@gmail.com.
authors.forename: Luciana Relly
authors.initials: LR
authors.lastname: Bertolini
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D056947:Enzyme Replacement Therapy; D005260:Female; D005776:Gaucher Disease / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D005962:Glucosylceramidase / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D005963:Glucosylceramides / Q000378:metabolism; D006041:Goats / Q000235:genetics; D006801:Humans; D008321:Mammary Glands, Animal / Q000201:enzymology; D008892:Milk / Q000378:metabolism; D011994:Recombinant Proteins / Q000008:administration & dosage*
chemical_list: D005963:Glucosylceramides; D011994:Recombinant Proteins; D005962:Glucosylceramidase
keywords: Adenovirus; Gaucher disease; Glucocerebrosidase; Glycosylation; Milk; Recombinant protein