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Transient Expression of Functional Glucocerebrosidase for Treatment of Gaucher’s Disease in the Goat Mammary Gland

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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.

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Acknowledgments

This work was supported by grants from FINEP/MCT/Brazil. K.C.S. Tavares was supported by a CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) fellowship from the Brazilian Government.

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Authors and Affiliations

  1. Molecular and Development Biology Laboratory, Experimental Biology Center, University of Fortaleza (UNIFOR), Fortaleza, CE, Brazil

    Kaio Cesar Simiano Tavares, Cícera Regina Lazzarotto, Saul Gaudencio Neto, Igor de Sá Carneiro, Felipe Ledur Ongaratto, Luís Henrique de Aguiar, Carlos Enrique Mendez Calderón, Marcelo Bertolini & Luciana Relly Bertolini

  2. Quatro G Pesquisa & Desenvolvimento Ltda., Tecnopuc, Porto Alegre, RS, Brazil

    Ana Christina de Oliveira Dias, Antônio Frederico Michel Pinto, Diogenes Santiago Santos & Jocelei Maria Chies

  3. Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción (Udec), Concepción, Chile

    Jorge Roberto Toledo

  4. Department of Physiopathology, Faculty of Veterinary Sciences, Universidad de Concepción (Udec), Chillán, Chile

    Fidel Ovidio Castro

  5. School of Veterinary Medicine, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil

    Marcelo Bertolini

  6. Biotechnology and Genetic Engineering Lab, School of Pharmacy, Pontifical Catholic University of Rio Grande do Sul (PUCRS), Porto Alegre, RS, Brazil

    Luciana Relly Bertolini

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  1. Kaio Cesar Simiano Tavares
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Correspondence to Luciana Relly Bertolini.

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All animal care and use were conducted in strict accordance with the Animal Research Committee guidelines of University of Fortaleza.

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Tavares, K.C.S., Dias, A.C.d.O., Lazzarotto, C.R. et al. Transient Expression of Functional Glucocerebrosidase for Treatment of Gaucher’s Disease in the Goat Mammary Gland. Mol Biotechnol 58, 47–55 (2016). https://doi.org/10.1007/s12033-015-9902-1

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