α-Galactosidase A-Tat Fusion Enhances Storage Reduction in Hearts and Kidneys of Fabry Mice

Koji Higuchi,Makoto Yoshimitsu,Xiaoxin Guo,Jeffrey A Medin,Chuwa Tei,Xin Fan,Toshihiro Takenaka,Jennifer Yen,Vanessa I Rasaiah

doi:10.2119/molmed.2009.00163

Koji Higuchi, Makoto Yoshimitsu + Show 7 more

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https://doi.org/10.2119/molmed.2009.00163

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Abstract

The protein transduction domain from human immunodeficiency virus (HIV) Tat allows proteins to penetrate the cell membrane. Enhanced cellular uptake of therapeutic proteins could benefit a number of disorders. This is especially true for lysosomal storage disorders (LSDs) where enzyme replacement therapy (ERT) and gene therapy have been developed. We developed a novel recombinant lentiviral vector (LV) that engineers expression of alpha-galactosidase A (alpha-gal A)-Tat fusion protein for correction of Fabry disease, the second-most prevalent LSD with manifestations in the brain, kidney and heart. In vitro experiments confirmed mannose-6-phosphate independent uptake of the fusion factor. Next, concentrated therapeutic LV was injected into neonatal Fabry mice. Analysis of tissues at 26 wks demonstrated similar alpha-gal A enzyme activities but enhanced globotriaosylceramide (Gb3) reduction in hearts and kidneys compared with the alpha-gal A LV control. This strategy might advance not only gene therapy for Fabry disease and other LSDs, but also ERT, especially for cardiac Fabry disease.

Highlights

Fabry disease is a lysosomal storage disorder (LSD) caused by a deficiency of α-galactosidase A (α-gal A; EC 3.2.1.22) activity [1], one of the lysosomal hydrolases
While α-gal A activity was completely inhibited by added M6P in the lentiviral vector (LV)/α-gal A treated group, α-gal A activity was only partially diminished by added M6P in LV/α-gal A-Tat treated group
In Experiment 1, there were no significant differences in α-gal A activity in most organs between LV/α-gal A and LV/α-gal A-Tat treated group even though we found the tendency of α-gal A activity to be higher in LV/α-gal A-Tat group in liver, heart, kidney, bone marrow and lung (Table 1)

Summary

Introduction

Fabry disease is a lysosomal storage disorder (LSD) caused by a deficiency of α-galactosidase A (α-gal A; EC 3.2.1.22) activity [1], one of the lysosomal hydrolases. It is the second-most prevalent LSD and a model for the development of therapy for single gene defects. Enzyme replacement therapy (ERT) for Fabry disease is available and some improvements in clinical and pathological manifestations have been shown. We have been focused on developing gene therapy for Fabry disease [7,8,9,10,11]

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