Article
Investigation of a recombinant SMN protein delivery system to treat spinal muscular atrophy
Translational Neuroscience
(2014)
Abstract
Spinal muscular atrophy (SMA), the most common genetic cause of infant death, is a neurodegenerative disorder
affecting motor neurons. SMA results from a loss in full-length survival of motor neuron (SMN) protein due to
deletions/mutations in the SMN1 gene. In this study, we assessed the ability of cell-penetrating peptides (CPP) to
deliver recombinant SMN protein to cultured neurons as a prelude for a potential therapeutic to treat SMA. Firstly,
we confirmed that E. coli produced recombinant GFP protein fused to TAT (YGRKKRRQRRR; TAT-GFP) transduced
rat cortical neurons in a concentration dependent manner. However, due to low yields of recombinant TATSMN
protein obtainable from E. coli, we investigated the potential of a modified TAT (TATκ: YARKAARQARA) or
R9 (RRRRRRRRR) peptide downstream of the fibronectin (FIB) secretory signal peptide to generate recombinant
CPP-fused SMN protein. While U251 cells transduced with an adenoviral vector expressing CMV-FIB-TATκ-SMN
secreted recombinant TATκ-SMN protein, we did not detect TATκ-SMN protein transduction of cortical neurons.
Further, purified TATκ-SMN was unable to transduce SH-SY5Y cells, nor block apoptosis following LY294002
treatment of these cells. Our findings indicate that TATκ is not a suitable CPP to deliver SMN protein to neurons.
Nonetheless, we have developed a novel method to generate full-length recombinant SMN protein using
a mammalian expression system, which can be used to explore the application of other CPPs to deliver SMN
protein as a treatment for SMA.
Disciplines
Publication Date
2014
DOI
10.2478/s13380-014-0201-2
Citation Information
Anderton, R., Meloni, B., Mastaglia, F., et al. (2014). Investigation of a recombinant SMN protein delivery system to treat spinal muscular atrophy. Translational Neuroscience, 5(1), 8-16. DOI: 10.2478/s13380-014-0201-2