Tài liệu Báo cáo khoa học: Treatment of neutral glycosphingolipid lysosomal storage diseases via

Treatment of neutral glycosphingolipid lysosomal storage

diseases via inhibition of the ABC drug transporter, MDR1

Cyclosporin A can lower serum and liver globotriaosyl ceramide

levels in the Fabry mouse model

Michael Mattocks1

, Maria Bagovich1

, Maria De Rosa1,4, Steve Bond2

, Beth Binnington1

,

Vanessa I. Rasaiah2

, Jeffrey Medin2,3 and Clifford Lingwood1,4,5

1 Research Institute, The Hospital for Sick Children, Toronto, Canada

2 Ontario Cancer Institute, University Health Network, Toronto, Canada

3 Department of Medical Biophysics, University of Toronto, Canada

4 Department of Laboratory Medicine and Pathology, University of Toronto, Canada

5 Department of Biochemistry, University of Toronto, Canada

The lysosomal storage diseases (LSD) are genetic defi￾ciencies in glycoconjugate catabolism, each due to a

lack of a specific lysosomal sugar hydrolase or its acti￾vator protein [1]. The (mainly neurological) symptoms

are due to the intracellular accumulation of the

enzyme substrate. In the ‘glycosphingolipidoses’, this

Keywords

enzyme replacement therapy; Gaucher

disease; a-galactosidase; glucosyl ceramide

translocase; HUS model

Correspondence

C. Lingwood, Research Institute, The

Hospital for Sick Children, Toronto, Ontario

M5G 1X8, Canada

Fax: +416 813 5993

Tel: +416 813 5998

E-mail: [email protected]

(Received 20 January 2006, revised 2 March

2006, accepted 10 March 2006)

doi:10.1111/j.1742-4658.2006.05223.x

We have shown that the ABC transporter, multiple drug resistance

protein 1 (MDR1, P-glycoprotein) translocates glucosyl ceramide from

the cytosolic to the luminal Golgi surface for neutral, but not acidic, gly￾cosphingolipid (GSL) synthesis. Here we show that the MDR1 inhibitor,

cyclosporin A (CsA) can deplete Gaucher lymphoid cell lines of accumu￾lated glucosyl ceramide and Fabry cell lines of globotriaosyl ceramide

(Gb3), by preventing de novo synthesis. In the Fabry mouse model, Gb3 is

increased in the heart, liver, spleen, brain and kidney. The lack of renal

glomerular Gb3 is retained, but the number of verotoxin 1 (VT1)-staining

renal tubules, and VT1 tubular targeting in vivo, is markedly increased in

Fabry mice. Adult Fabry mice were treated with a-galactosidase (enzyme￾replacement therapy, ERT) to eliminate serum Gb3 and lower Gb3 levels in

some tissues. Serum Gb3 was monitored using a VT1 ELISA during a

post-ERT recovery phase ± biweekly intra peritoneal CsA. After 9 weeks,

tissue Gb3 content and localization were determined using VT1 ⁄TLC over￾lay and histochemistry. Serum Gb3 recovered to lower levels after CsA

treatment. Gb3 was undetected in wild-type liver, and the levels of Gb3

(but not gangliosides) in Fabry mouse liver were significantly depleted by

CsA treatment. VT1 liver histochemistry showed Gb3 accumulated in

Kupffer cells, endothelial cell subsets within the central and portal vein and

within the portal triad. Hepatic venule endothelial and Kupffer cell VT1

staining was considerably reduced by in vivo CsA treatment. We conclude

that MDR1 inhibition warrants consideration as a novel adjunct treatment

for neutral GSL storage diseases.

Abbreviations

BSA, bovine serum albumin; CsA, cyclosporin A; ERT, enzyme replacement therapy; Gb3, globotriaosyl ceramide; GlcCer, glucosyl ceramide;

GSL, glycosphingolipid; HUS, hemolytic uremic syndrome; LacCer, lactosyl ceramide; LSD, lysosomal storage disease; MDR1, multiple drug

resistance protein 1 (P-glycoprotein); NGS, normal goat serum; VT1, verotoxin 1; TLC, thin layer chromatogram.

2064 FEBS Journal 273 (2006) 2064–2075 ª 2006 The Authors Journal compilation ª 2006 FEBS