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. 2024 Dec 17;13(1):63.
doi: 10.1186/s40035-024-00455-4.

Recombinant SMN protein synergizes with spinal muscular atrophy therapy to counteract pathological motor neuron phenotypes

Liliana Brambilla  1 , Chiara F Valori  1 , Giulia Guidotti  1 , Francesca Martorana  1 , Claudia Sulmona  1 , Lisa Benedetta De Martini  1 , Anselmo Canciani  2 , Marco Fumagalli  2 , Francesca Talpo  3 , Gerardo Biella  3 , Elisa Di Pasquale  4   5 , Claudio Iacobucci  6 , Federico Forneris  2   7 , Haiyan Zhou  8 , Daniela Rossi  9
Affiliations

Recombinant SMN protein synergizes with spinal muscular atrophy therapy to counteract pathological motor neuron phenotypes

Liliana Brambilla et al. Transl Neurodegener. .
No abstract available

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Conflict of interest statement

Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: D.R. and L.B. are inventors on patent applications filed by ICSM related to TAT-conjugated peptides/proteins.

Figures

Fig. 1
Fig. 1
a Schematic presentation of recombinant TAT-flSMN protein topology and mass-spectrometry peak profile obtained from tryptic digestion of the sample. Inset shows the peptide sequence coverage (underlined) on the entire protein sequence. b Images show NSC-34 cells treated with 100 nM unlabelled (left) or Fluorescein-labelled TAT-flSMN (right, green). Nuclei were stained with Hoechst 33342 (blue). Scale bar, 50 μm. c Representative immunoblots (left) of SMN and β-actin proteins in the A.1 CTRL NSC-34 cells and the GM.5 and C.2 SMN-knockdown NSC-34 cell lines. Bands were quantified and SMN/β-actin ratios were calculated for normalization (right) (mean ± SEM; n = 3–4; *P < 0.05 and ***P < 0.001, Bonferroni-corrected one-way ANOVA). d Percentage of cells without neurites (left) and the mean length of the longest neurite per cell (right) in GM.5 (green) and C.2 (red) SMN-knockdown cells in the absence or presence of increasing concentrations of TAT-flSMN (mean ± SEM; n = 6–7; *P < 0.05, **P < 0.01, ***P < 0.0001, Bonferroni-corrected one–way ANOVA). e Percentage of cells with condensed nuclei (left) or showing active caspase 3 (right) in GM.5 (green) and C.2 (red) SMN-knockdown cells treated as in (d) (mean ± SEM; n = 6; *P < 0.05, **P < 0.001, ***P < 0.0001, Bonferroni-corrected one–way ANOVA). f Representative image of iPSC-derived MNs from a SMA patient stained with Hoechst 33342 and immunolabeled for beta III-tubulin (TUJ1) and choline acetyltransferase (ChAT). Scale bar, 50 µm. g Analyses of the average neurite length per cell in SMA iPSC-derived MNs treated with 10 nM TAT-flSMN, 0.5 μM PMO25, or 10 nM TAT-flSMN + 0.5 μM PMO25 (mean ± SEM; n = 4; *P < 0.05, **P < 0.01, ***P < 0.0001; Bonferroni-corrected one-way ANOVA). h Analysis of SMA iPSC-derived MNs with condensed nuclei or TUNEL+ staining upon treatment as in (g) (mean ± SEM; n = 6; *P < 0.05, **P < 0.01, ***P < 0.0001, Bonferroni-corrected one–way ANOVA)
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References

    1. Yeo CJJ, Tizzano EF, Darras BT. Challenges and opportunities in spinal muscular atrophy therapeutics. Lancet Neurol. 2024;23(2):205–18. - DOI - PubMed
    1. Dangouloff T, Servais L. Clinical evidence supporting early treatment of patients with spinal muscular atrophy: current perspectives. Ther Clin Risk Manag. 2019;15:1153–61. - DOI - PMC - PubMed
    1. Martorana F, Brambilla L, Valori CF, Bergamaschi C, Roncoroni C, Aronica E, et al. The BH4 domain of Bcl-X(L) rescues astrocyte degeneration in amyotrophic lateral sclerosis by modulating intracellular calcium signals. Hum Mol Genet. 2012;21(4):826–40. - DOI - PubMed
    1. Lorson CL, Strasswimmer J, Yao JM, Baleja JD, Hahnen E, Wirth B, et al. SMN oligomerization defect correlates with spinal muscular atrophy severity. Nat Genet. 1998;19(1):63–6. - DOI - PubMed
    1. Kariya S, Park GH, Maeno-Hikichi Y, Leykekhman O, Lutz C, Arkovitz MS, et al. Reduced SMN protein impairs maturation of the neuromuscular junctions in mouse models of spinal muscular atrophy. Hum Mol Genet. 2008;17(16):2552–69. - DOI - PMC - PubMed

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