Reversal of Neuroinflammation in Novel GS Model Mice by Single I.c.v. Administration of CHO-derived RhCTSA Precursor Protein

Overview

Journal Mol Ther Methods Clin Dev

Publisher Cell Press

Date 2022 May 16

PMID 35573044

Authors

Yuto Horii

Toshiki Iniwa

Masayoshi Onitsuka

Jun Tsukimoto

Yuki Tanaka

Hironobu Ike

Yuri Fukushi

Haruna Ando

Yoshie Takeuchi

So-Ichiro Nishioka

Daisuke Tsuji

Mariko Ikuo

Naoshi Yamazaki

Yoshiharu Takiguchi

Naozumi Ishimaru

Kohji Itoh

Affiliations

Soon will be listed here.

Abstract

Galactosialidosis (GS) is a lysosomal cathepsin A (CTSA) deficiency. It associates with a simultaneous decrease of neuraminidase 1 (NEU1) activity and sialylglycan storage. Central nervous system (CNS) symptoms reduce the quality of life of juvenile/adult-type GS patients, but there is no effective therapy. Here, we established a novel GS model mouse carrying homozygotic mutation causing partial exon 6 skipping with concomitant deficiency of Ctsa/Neu1. The GS mice developed juvenile/adult GS-like symptoms, such as gargoyle-like face, edema, proctoprosia due to sialylglycan accumulation, and neurovisceral inflammation, including activated microglia/macrophage appearance and increase of inflammatory chemokines. We produced human CTSA precursor proteins (proCTSA), a homodimer carrying terminal mannose 6-phosphate (M6P)-type N-glycans. The CHO-derived proCTSA was taken up by GS patient-derived fibroblasts via M6P receptors and delivered to lysosomes. Catalytically active mature CTSA showed a shorter half-life due to intralysosomal proteolytic degradation. Following single i.c.v. administration, proCTSA was widely distributed, restored the Neu1 activity, and reduced the sialylglycans accumulated in brain regions. Moreover, proCTSA suppressed neuroinflammation associated with reduction of activated microglia/macrophage and up-regulated Mip1α. The results show therapeutic effects of intracerebrospinal enzyme replacement utilizing CHO-derived proCTSA and suggest suppression of CNS symptoms.

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References

Kiriyama K, Itoh K . Glycan Recognition and Application of P-Type Lectins. Methods Mol Biol. 2020; 2132:267-276. DOI: 10.1007/978-1-0716-0430-4_26. View

Sakuraba H, Aoyagi T, Suzuki Y . Galactosialidosis (beta-galactosidase-neuraminidase deficiency): a possible role of serine-thiol proteases in the degradation of beta-galactosidase molecules. Clin Chim Acta. 1982; 125(3):275-82. DOI: 10.1016/0009-8981(82)90257-1. View

Leimig T, Mann L, del Pilar Martin M, Bonten E, Persons D, Knowles J . Functional amelioration of murine galactosialidosis by genetically modified bone marrow hematopoietic progenitor cells. Blood. 2002; 99(9):3169-78. DOI: 10.1182/blood.v99.9.3169. View

Thomas C, Schiedner G, Kochanek S, Castro M, Lowenstein P . Preexisting antiadenoviral immunity is not a barrier to efficient and stable transduction of the brain, mediated by novel high-capacity adenovirus vectors. Hum Gene Ther. 2001; 12(7):839-46. DOI: 10.1089/104303401750148829. View

Martins C, Hulkova H, Dridi L, Dormoy-Raclet V, Grigoryeva L, Choi Y . Neuroinflammation, mitochondrial defects and neurodegeneration in mucopolysaccharidosis III type C mouse model. Brain. 2015; 138(Pt 2):336-55. PMC: 4306821. DOI: 10.1093/brain/awu355. View

Caciotti A, Catarzi S, Tonin R, Lugli L, Rodriguez Perez C, Michelakakis H . Galactosialidosis: review and analysis of CTSA gene mutations. Orphanet J Rare Dis. 2013; 8:114. PMC: 3737020. DOI: 10.1186/1750-1172-8-114. View

Jackman H, Tan F, Tamei H, Li X, Skidgel R, Erdos E . A peptidase in human platelets that deamidates tachykinins. Probable identity with the lysosomal "protective protein". J Biol Chem. 1990; 265(19):11265-72. View

Durand P, Gatti R, Cavalieri S, BORRONE C, Tondeur M, Michalski J . Sialidosis (mucolipidosis I). Helv Paediatr Acta. 1977; 32(4-5):391-400. View

Oheda Y, Kotani M, Murata M, Sakuraba H, Kadota Y, Tatano Y . Elimination of abnormal sialylglycoproteins in fibroblasts with sialidosis and galactosialidosis by normal gene transfer and enzyme replacement. Glycobiology. 2005; 16(4):271-80. DOI: 10.1093/glycob/cwj069. View

10.

Luu A, Wong C, Agrawal V, Wise N, Handyside B, Lo M . Intermittent enzyme replacement therapy with recombinant human β-galactosidase prevents neuraminidase 1 deficiency. J Biol Chem. 2020; 295(39):13556-13569. PMC: 7521647. DOI: 10.1074/jbc.RA119.010794. View

11.

Galjart N, Gillemans N, Harris A, van der Horst G, Verheijen F, GALJAARD H . Expression of cDNA encoding the human "protective protein" associated with lysosomal beta-galactosidase and neuraminidase: homology to yeast proteases. Cell. 1988; 54(6):755-64. DOI: 10.1016/s0092-8674(88)90999-3. View

12.

Chen J, Luu A, Wise N, De Angelis R, Agrawal V, Mangini L . Intracerebroventricular enzyme replacement therapy with β-galactosidase reverses brain pathologies due to GM1 gangliosidosis in mice. J Biol Chem. 2019; 295(39):13532-13555. PMC: 7521651. DOI: 10.1074/jbc.RA119.009811. View

13.

Itoh K, Takiyama N, Kase R, Kondoh K, Sano A, Oshima A . Purification and characterization of human lysosomal protective protein expressed in stably transformed Chinese hamster ovary cells. J Biol Chem. 1993; 268(2):1180-6. View

14.

Slama T, Garbade S, Kolker S, Hoffmann G, Ries M . Quantitative natural history characterization in a cohort of 142 published cases of patients with galactosialidosis-A cross-sectional study. J Inherit Metab Dis. 2019; 42(2):295-302. DOI: 10.1002/jimd.12010. View

15.

Seo J, Kosuga M, Hamazaki T, Shintaku H, Okuyama T . Impact of intracerebroventricular enzyme replacement therapy in patients with neuronopathic mucopolysaccharidosis type II. Mol Ther Methods Clin Dev. 2021; 21:67-75. PMC: 7957024. DOI: 10.1016/j.omtm.2021.02.018. View

16.

Itoh K, Takiyama N, Nagao Y, Oshima A, Sakuraba H, Potier M . Acid carboxypeptidase deficiency in galactosialidosis. Jinrui Idengaku Zasshi. 1991; 36(2):171-7. DOI: 10.1007/BF01876581. View

17.

Rombach S, Smid B, Linthorst G, Dijkgraaf M, Hollak C . Natural course of Fabry disease and the effectiveness of enzyme replacement therapy: a systematic review and meta-analysis: effectiveness of ERT in different disease stages. J Inherit Metab Dis. 2014; 37(3):341-52. DOI: 10.1007/s10545-014-9677-8. View

18.

Kotani M, Yamada H, Sakuraba H . Cytochemical and biochemical detection of intracellularly accumulated sialyl glycoconjugates in sialidosis and galactosialidosis fibroblasts with Macckia amurensis. Clin Chim Acta. 2004; 344(1-2):131-5. DOI: 10.1016/j.cccn.2004.02.032. View

19.

Zhou X, Morreau H, Rottier R, Davis D, Bonten E, Gillemans N . Mouse model for the lysosomal disorder galactosialidosis and correction of the phenotype with overexpressing erythroid precursor cells. Genes Dev. 1995; 9(21):2623-34. DOI: 10.1101/gad.9.21.2623. View

20.

Wraith J, Scarpa M, Beck M, Bodamer O, De Meirleir L, Guffon N . Mucopolysaccharidosis type II (Hunter syndrome): a clinical review and recommendations for treatment in the era of enzyme replacement therapy. Eur J Pediatr. 2007; 167(3):267-77. PMC: 2234442. DOI: 10.1007/s00431-007-0635-4. View