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In vitro correction of ARSA deficiency in human skin fibroblasts from Metachromatic Leukodystrophy patients after treatment with microencapsulated recombinant cells

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Abstract

Metachromatic leukodystrophy (MLD) is an autosomal recessive disorder due to arylsulfatase A (ARSA) deficiency that affects primarily the central nervous system. Ongoing treatments include enzyme replacement therapy and bone marrow transplantation, both limited in their effects due to the blood-brain barrier. An alternative approach would be the in situ implantation of encapsulated cells over expressing ARSA. Based on that, we tested the ability of encapsulated BHK cells over expressing ARSA to correct the enzyme deficiency in MLD patients' fibroblasts. Three groups were analyzed: fibroblasts treated with ARSA-over expressing BHK cells (rBHK) trapped in alginate capsules (capsules group), fibroblasts treated with supernatant of non-encapsulated rBHK (uptake control) and fibroblasts treated with empty capsules (empty group). Untreated and normal fibroblasts were used as controls. rBHK obtained by clone selection after non-viral transfection with pTARSA-CMV2. ARSA activity was measured after 1, 2, 3 and 4 weeks of treatment and β-gal was used as reference enzyme. Statistical analysis was performed using ANOVA and Tukey's test. Normal fibroblasts showed ARSA activity of 23.9 + /− 2.01 nmol/h/mg of protein, whereas untreated MLD fibroblasts had the low ARSA activity (2.22 + /− 0.17). In the empty group, ARSA activity was equal to that of untreated fibroblasts (2.71 + /− 0.34). Capsules and uptake control groups showed higher enzymatic activity levels, compared to MLD untreated, 23.42 + /− 6.39 and 42.35 + /− 5.20, respectively (p < 0.01 for all groups). Encapsulated rBHK clones show potential as a new therapeutic strategy for the treatment of MLD, reaching normal enzyme levels in human MLD fibroblasts.

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References

  • Altarescu G, Hill S, Wiggs E, Jeffries N, Kreps C, Parker CC, Brady RO, Barton NW, Schiffmann R (2001) The efficacy of enzyme replacement therapy in patients with chronic neuronopathic Gaucher's disease. J Pediatr 138:539–547

    Article  PubMed  CAS  Google Scholar 

  • Andersson S, Davis DL, Dahlbäck H, Jörnvall H, Russell DW (1989) Cloning, structure, and expression of the mitochondrial cytochrome P-450 sterol 26-hydroxylase, a bile acid biosynthetic enzyme. J Biol Che 264:8222–8229

    CAS  Google Scholar 

  • Anson DS, Bielicki J, Hopwood JJ (1992) Correction of mucopolysaccharidosis type I fibroblasts by retroviral-mediated transfer of the human alpha-L-iduronidase gene. Hum Gene Ther 3(4):371–379

    Article  PubMed  CAS  Google Scholar 

  • Bloch J, Bachoud-Lévi AC, Déglon N, Lefaucheur JP, Winkel L, Palfi S, Nguyen JP, Bourdet C, Gaura V, Remy P, Brugières P, Boisse MF, Baudic S, Cesaro P, Hantraye P, Aebischer P, Peschanski M (2004) Neuroprotective gene therapy for Huntington's disease, using polymer-encapsulated cells engineered to secrete human ciliary neurotrophic factor: results of a phase I study. Hum Gene Ther 15(10):968–975

    Article  PubMed  CAS  Google Scholar 

  • Brooks AR, Harkins RN, Wang P, Qian HS, Liu P, Rubanyi GM (2004) Transcriptional silencing is associated with extensive methylation of the CMV promoter following adenoviral gene delivery to muscle. J Gene Med 6(4):395–404

    Article  PubMed  CAS  Google Scholar 

  • Capotondo A, Cesani M, Pepe S, Fasano S, Gregori S, Tononi L, Venneri MA, Brambilla R, Quattrini A, Ballabio A, Cosma MP, Naldini L, Biffi A (2007) Safety of arylsulfatase A overexpression for gene therapy of metachromatic leukodystrophy. Hum Gene Ther 18:821–836

    Article  PubMed  CAS  Google Scholar 

  • Cheng SH, Smith AE (2003) Gene therapy progress and prospects: gene therapy of lysosomal storage disorders. Gene Ther 10(16):1275–1281

    Article  PubMed  CAS  Google Scholar 

  • Consiglio A, Martino S, Dolcetta D, Cusella G, Conese M, Marchesini S, Benaglia G, Wrabetz L, Orlacchio A, Déglon N, Aebischer P, Severini GM, Bordignon C (2007) Metabolic correction in oligodendrocytes derived from metachromatic leukodystrophy mouse model by using encapsulated recombinant myoblasts. J Neurol Sci 255:7–16

    Article  PubMed  CAS  Google Scholar 

  • Consiglio A, Quattrini A, Martino S, Bensadoun JC, Dolcetta D, Trojani A, Benaglia G, Marchesini S, Cestari V, Oliverio A, Bordignon C, Naldini L (2001) In vivo gene therapy of metachromatic leukodystrophy by lentiviral vectors: correction of neuropathology and protection against learning impairments in affected mice. Nat Med 7:310–316

    Article  PubMed  CAS  Google Scholar 

  • Desnick RJ, Schuchman EH (2002) Enzyme replacement and enhancement therapies: lessons from lysosomal disorders. Nat Rev Genet 3:954–966

    Article  PubMed  CAS  Google Scholar 

  • Friso A, Tomanin R, Alba S, Gasparotto N, Puicher EP, Fusco M, Hortelano G, Muenzer J, Marin O, Zacchello F, Scarpa M (2005) Reduction of GAG storage in MPS II mouse model following implantation of encapsulated recombinant myoblasts. J Gene Med 7:1482–1491

    Article  PubMed  CAS  Google Scholar 

  • Grassi G, Maccaroni P, Meyer R, Kaiser H, D, Ambrosio E, Pascale E, Grassi M, Kuhn A, Di Nardo P, Kandolf R, Küpper JH (2003) Inhibitors of DNA methylation and histone deacetylation activate cytomegalovirus promoter-controlled reporter gene expression in human glioblastoma cell line U87. Carcinogenesis 24(10):1625–1635

    Article  PubMed  CAS  Google Scholar 

  • Kakkis ED, Schuchman E, He X, Wan Q, Kania S, Wiemelt S, Hasson CW, O’Malley T, Weil MA, Aguirre GA, Brown DE, Haskins ME (2001) Enzyme replacement therapy in feline mucopolysaccharidosis I. Mol Genet Metab 72:199–208

    Article  PubMed  CAS  Google Scholar 

  • Kamensky E, Philippart M, Cancilla P, Frommes SP (1973) Cultured skin fibroblasts in storage disorders. An analysis of ultrastructural features. Am J Pathol 73(1):59–80

    PubMed  CAS  Google Scholar 

  • Kuijlen JM, de Haan BJ, Helfrich W, de Boer JF, Samplonius D, Mooij JJ, de Vos P (2006) The efficacy of alginate encapsulated CHO-K1 single chain-TRAIL producer cells in the treatment of brain tumors. J Neurooncol 78:31–39

    Article  PubMed  CAS  Google Scholar 

  • Lee-Vaupel M, Conzelmann E (1987) A simple chromogenic assay for arylsulfatase A. Clin Chim Acta 164:171–180

    Article  PubMed  CAS  Google Scholar 

  • Lindvall O, Wahlberg LU (2008) Encapsulated cell biodelivery of GDNF: a novel clinical strategy for neuroprotection and neuroregeneration in Parkinson's disease? Exp Neurol 209:82–88

    Article  PubMed  CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  • Maass A, Langer SJ, Oberdorf-Maass S, Bauer S, Neyses L, Leinwand LA (2003) Rational promoter selection for gene transfer into cardiac cells. J Mol Cell Cardiol 35(7):823–831

    Article  PubMed  CAS  Google Scholar 

  • Mabe-Santana P (2006) La enfermedad de Krabbe y la leucodistrofia metacromática. In: Sanjurjo P, Baldellou A (eds), Diagnóstico y tratamiento de las enfermedades metabólicas hereditárias, Ed. Ergon, pp 639–650

  • Marcus J, Honigbaum S, Shroff S, Honke K, Rosenbluth J, Dupree JL (2006) Sulfatide is essencial for the maintenance of CNS myelin and axon struture. Glia 53:372–381

    Article  PubMed  CAS  Google Scholar 

  • Matzner U, Herbst E, Hedayati KK, Lüllmann-Rauch R, Wessig C, Schröder S, Eistrup C, Möller C, Fogh J, Gieselmann V (2005) Enzyme replacement improves nervous system pathology and function in a mouse model for metachromatic leukodystrophy. Hum Mol Genet 14:1139–1152

    Article  PubMed  CAS  Google Scholar 

  • Molinas MMB (2005) Transient Transfection of HEK293 Cells in Suspension: Process Characterization and Optimization by Applying Invasive Nucleotide and Non-invasive Electronic Nose Technology, Doctoral Thesis

  • Nakama H, Ohsugi K, Otsuki T, Date I, Kosuga M, Okuyama T, Sakuragawa N (2006) Encapsulation cell therapy for mucopolysaccharidosis type VII using genetically engineered immortalized human amniotic epithelial cells. Tohoku J Exp Med 209:23–32

    Article  PubMed  CAS  Google Scholar 

  • Nelson JA, Reynolds-Kohler C, Smith BA (1987) Negative and positive regulation by a short segment in the 5′-flanking region of the human cytomegalovirus major immediate-early gene. Mol Cell Biol 7:4125–4129

    PubMed  CAS  Google Scholar 

  • Passini MA, Wolfe JH (2001) Widespread gene delivery and structure-specific patterns of expression in the brain after intraventricular injections of neonatal mice with an adeno-associated virus vector. J Virol 75:12382–12392

    Article  PubMed  CAS  Google Scholar 

  • Passini MA, Macauley SL, Huff MR, Taksir TV, Bu J, Wu IH, Piepenhagen PA, Dodge JC, Shihabuddin LS, O’Riordan CR, Schuchman EH, Stewart GR (2005) AAV vector-mediated correction of brain pathology in a mouse model of Niemann-Pick A disease. Mol Ther 11:754–762

    Article  PubMed  CAS  Google Scholar 

  • Pérez-Calvo J, Giraldo P, Pastores GM, Fernández-Galán M, Martín-Nuñez G, Pocoví M (2003) Extended interval between enzyme therapy infusions for adult patients with Gaucher's disease type 1. J Postgrad Med 49:127–131

    PubMed  Google Scholar 

  • QIAGEN (1999) The QIAGEN transfection resource book

  • Ross CJ, Ralph M, Chang PL (2000) Somatic gene therapy for a neurodegenerative disease using microencapsulated recombinant cells. Exp Neurol 166:276–286

    Article  PubMed  CAS  Google Scholar 

  • Saravanan K, Büssow H, Weiler N, Gieselmann V, Franken S (2007) Spontaneously immortalized Schwann cell line to study the molecular aspects of metachromatic leukodystrophy. Neurosci Methods 161(2):223–233

    Article  CAS  Google Scholar 

  • Schott I, Hartmann D, Gieselmann V, Lüllmann-Rauch R (2001) Sulfatide storage in visceral organs of arylsulfatase A-deficient mice. Virchows Arch 439(1):90–96

    Article  PubMed  CAS  Google Scholar 

  • Stein C, Gieselmann V, Kreysing J, Schimidt B, Pohlmann R, Waheed A, Meyer HE, O’brien JS, Von Figura K (1989) Cloning and expression of human arylsulfatase A. J Biol Chem 264:1252–1259

    PubMed  CAS  Google Scholar 

  • Suzuki K (1977) Globoid cell leukodystrophy (Krabbe disease) and GM1 gangliosidosis. In: Glew RH, Petersen SP (eds) Pratical enzymology of the sphingolipidosis. Alan R. Liss, New York

    Google Scholar 

  • Teschendorf C, Warrington KH, Siemann DW, Muzyczka N (2002) Comparison of the EF-1 alpha and the CMV promoter for engineering stable tumor cell lines using recombinant adeno-associated virus. Anticancer Res 22(6A):3325–3330

    PubMed  CAS  Google Scholar 

  • Visted T, Bjerkvig R, Enger PO (2001) Cell encapsulation technology as a therapeutic strategy for CNS malignancies. Neuro Oncol 3:201–210

    Article  PubMed  CAS  Google Scholar 

  • Von Figura K, Gieselmann V, Jaeken J (2001) Metachromatic leukodystrophy. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic and molecular bases of inherited disease. McGraw-Hill, New York, pp 3695–3724

    Google Scholar 

  • Wittke D, Hartmann D, Gieselmann V, Lüllmann-Rauch R (2004) Lysosomal sulfatide storage in the brain of arylsulfatase A-deficient mice: cellular alterations and topographic distribution. Acta Neuropathol 108(4):261–271

    Article  PubMed  CAS  Google Scholar 

  • Wraith JE (2001) Enzyme replacement therapy in mucopolysaccharidosis type I: progress and emerging difficulties. J Inherit Metab Dis 24:245–250

    Article  PubMed  CAS  Google Scholar 

  • Yoshida H, Date I, Shingo T, Fujiwara K, Miyoshi Y, Furuta T, Ohmoto T (1999) Evaluation of reaction of primate brain to grafted PC12 cells. Cell Transplant 8:427–430

    PubMed  CAS  Google Scholar 

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Acknowledgments

The authors are grateful to Dr. Volkmar Gieselmann for kindly providing the pBEH HT14/CP8 vector with human ARSA cDNA. This research was supported by grants from Gene Therapy Network/Millenium Institute/MCT, CNPq, FIPE-HCPA and ONG Pela Vida.

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Authors and Affiliations

  1. Postgraduate Program in Genetics and Molecular Biology, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Valeska Lizzi Lagranha, Maria Luiza Saraiva-Pereira & Roberto Giugliani

  2. Postgraduate Program in Biochemistry, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Guilherme Baldo, Maria Luiza Saraiva-Pereira & Roberto Giugliani

  3. Gene Therapy Center/Research Center, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Valeska Lizzi Lagranha, Guilherme Baldo, Talita Giacomet de Carvalho, Ursula Matte & Roberto Giugliani

  4. Medical Genetics Service, Hospital de Clínicas de Porto Alegre, Poryo Alegre, RS, Brazil

    Maira Burin, Maria Luiza Saraiva-Pereira & Roberto Giugliani

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  1. Valeska Lizzi Lagranha
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  2. Guilherme Baldo
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  3. Talita Giacomet de Carvalho
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  6. Ursula Matte
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  7. Roberto Giugliani
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Correspondence to Valeska Lizzi Lagranha.

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Lagranha, V.L., Baldo, G., de Carvalho, T.G. et al. In vitro correction of ARSA deficiency in human skin fibroblasts from Metachromatic Leukodystrophy patients after treatment with microencapsulated recombinant cells. Metab Brain Dis 23, 469–484 (2008). https://doi.org/10.1007/s11011-008-9107-4

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  • DOI: https://doi.org/10.1007/s11011-008-9107-4

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