Human Recombinant Lysosomal β-Hexosaminidase A produced in P.pastoris efficiently reduced GM2 ganglioside accumulation in Tay- Sachs Disease

Author:

Inci Orhan Kerim1,Leal Andrés Felipe2,Ateş Nurselin1,Súarez Diego A.2,Espejo-Mojica Angela Johana2,Alméciga-Diaz Carlos Javier2,Seyrantepe Volkan1

Affiliation:

1. Izmir Institute of Technology

2. Pontificia Universidad Javeriana

Abstract

Abstract

Tay-Sachs disease is a progressive lysosomal storage disorder caused by genetic mutations in the HEXA gene encoding α-subunit of β-Hexosaminidase A, which leads to GM2 ganglioside accumulation, particularly in the central nervous system. Lysosomal GM2 ganglioside accumulation causes neuropathology and leads to premature cell death in Tay-Sachs patients, and there is no effective treatment yet. Previously, we produced the human recombinant lysosomal β-Hexosaminidase (rhHex-A) in the yeast Pichia pastoris, demonstrating the capacity to be internalized in different cell lines, and the reduction of stored GM2 gangliosides in patients´ fibroblasts and neural iPSCs. In this study, we use mouse fibroblasts and neuroglia from a novel murine model of Tay-Sachs disease and human Tay-Sachs patients’ neuroglia cells to demonstrate whether rhHex-A could reduce the GM2 accumulation. We treated the cells with rhHex-A to a final concentration of 100 nM, and the efficacy was evaluated using qRT-PCR and Immunocytochemical analysis after 24–72 h of incubation. Altogether, our results suggest that rhHex-A-produced in the yeast Pichia pastoris is a promising therapeutic to mitigate GM2 accumulation in Tay-Sachs cells.

Publisher

Springer Science and Business Media LLC

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