Limits...
Synergic prodegradative activity of Bicalutamide and trehalose on the mutant androgen receptor responsible for spinal and bulbar muscular atrophy.

Giorgetti E, Rusmini P, Crippa V, Cristofani R, Boncoraglio A, Cicardi ME, Galbiati M, Poletti A - Hum. Mol. Genet. (2014)

Bottom Line: Thus, (i) prevention of ARpolyQ nuclear localization, combined with (ii) an increased ARpolyQ cytoplasmic clearance, should reduce its detrimental activity.Using the antiandrogen Bicalutamide (Casodex(®)), which slows down AR activation and nuclear translocation, and the disaccharide trehalose, an autophagy activator, we found that, in motoneurons, the two compounds together reduced ARpolyQ insoluble forms with higher efficiency than that obtained with single treatments.Collectively, these data suggest that the combinatory use of Bicalutamide and trehalose is a novel approach to facilitate ARpolyQ clearance that has to be tested in other cell types target of SBMA (i.e. muscle cells) and in vivo in animal models of SBMA.

View Article: PubMed Central - PubMed

Affiliation: Sezione di Biomedicina ed Endocrinologia, Dipartimento di Scienze Farmacologiche e Biomolecolari (DiSFeB), Centro di Eccellenza sulle Malattie Neurodegenerative, Università degli Studi di Milano, Milano 20133, Italy Centro InterUniversitario sulle Malattie Neurodegenerative, Università degli Studi di Firenze, Genova e Roma Tor Vergata, Milano 20133, Italy Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA and.

Show MeSH

Related in: MedlinePlus

SQSTM1/p62 and LC3 distribution. (A and B) HRFM analysis (63× magnification) on NSC34 cells expressing GFP-AR.Q48 in the absence of (EtOH) or in the presence of 10 nm testosterone (T) and/or 100 nm Bicalutamide (Cas), with or without 100 mm trehalose treatment for 48 h. Endogenous SQSTM1/p62 (A) and LC3 (B) protein distribution and expression levels are shown in red. Nuclei were stained with DAPI (blue). Scale bar = 10 μm.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4262493&req=5

DDU419F5: SQSTM1/p62 and LC3 distribution. (A and B) HRFM analysis (63× magnification) on NSC34 cells expressing GFP-AR.Q48 in the absence of (EtOH) or in the presence of 10 nm testosterone (T) and/or 100 nm Bicalutamide (Cas), with or without 100 mm trehalose treatment for 48 h. Endogenous SQSTM1/p62 (A) and LC3 (B) protein distribution and expression levels are shown in red. Nuclei were stained with DAPI (blue). Scale bar = 10 μm.

Mentions: These results were further supported by the data obtained in IF microscopy analysis (Fig. 5) When cells were not treated with trehalose, SQSTM1/p62 shows an irregular and disorganized distribution, as shown in Figure 5A. In the presence of trehalose, SQSTM1/p62 expression levels notably increase and SQSTM1/p62 is homogeneously distributed into the entire cell cytoplasm. No changes in SQSTM1/p62 expression and distribution are induced by Bicalutamide (Cas) treatment. In the case of LC3 distribution analyzed in IF (Fig. 5B), we found that Bicalutamide (Cas) does not influence the levels or the overall punctate distribution of LC3-II induced by mutant ARpolyQ (both in the absence and presence of testosterone); conversely, trehalose treatment, which stimulates autophagy, enhances LC3-II levels which become exclusively present in its punctate distribution. As expected, similar levels and distribution of LC3-II are present in immortalized motoneurons expressing ARpolyQ and simultaneously treated with Bicalutamide (Cas) and trehalose (both in the absence and presence of testosterone). In all cases, Bicalutamide and trehalose alone or in combination are all capable to reduce the number of ARpolyQ aggregates formed after testosterone treatment. All together, the data strongly suggest the presence of active autophagy capable to remove the cytoplasmically retained ARpolyQ.Figure 5.


Synergic prodegradative activity of Bicalutamide and trehalose on the mutant androgen receptor responsible for spinal and bulbar muscular atrophy.

Giorgetti E, Rusmini P, Crippa V, Cristofani R, Boncoraglio A, Cicardi ME, Galbiati M, Poletti A - Hum. Mol. Genet. (2014)

SQSTM1/p62 and LC3 distribution. (A and B) HRFM analysis (63× magnification) on NSC34 cells expressing GFP-AR.Q48 in the absence of (EtOH) or in the presence of 10 nm testosterone (T) and/or 100 nm Bicalutamide (Cas), with or without 100 mm trehalose treatment for 48 h. Endogenous SQSTM1/p62 (A) and LC3 (B) protein distribution and expression levels are shown in red. Nuclei were stained with DAPI (blue). Scale bar = 10 μm.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4262493&req=5

DDU419F5: SQSTM1/p62 and LC3 distribution. (A and B) HRFM analysis (63× magnification) on NSC34 cells expressing GFP-AR.Q48 in the absence of (EtOH) or in the presence of 10 nm testosterone (T) and/or 100 nm Bicalutamide (Cas), with or without 100 mm trehalose treatment for 48 h. Endogenous SQSTM1/p62 (A) and LC3 (B) protein distribution and expression levels are shown in red. Nuclei were stained with DAPI (blue). Scale bar = 10 μm.
Mentions: These results were further supported by the data obtained in IF microscopy analysis (Fig. 5) When cells were not treated with trehalose, SQSTM1/p62 shows an irregular and disorganized distribution, as shown in Figure 5A. In the presence of trehalose, SQSTM1/p62 expression levels notably increase and SQSTM1/p62 is homogeneously distributed into the entire cell cytoplasm. No changes in SQSTM1/p62 expression and distribution are induced by Bicalutamide (Cas) treatment. In the case of LC3 distribution analyzed in IF (Fig. 5B), we found that Bicalutamide (Cas) does not influence the levels or the overall punctate distribution of LC3-II induced by mutant ARpolyQ (both in the absence and presence of testosterone); conversely, trehalose treatment, which stimulates autophagy, enhances LC3-II levels which become exclusively present in its punctate distribution. As expected, similar levels and distribution of LC3-II are present in immortalized motoneurons expressing ARpolyQ and simultaneously treated with Bicalutamide (Cas) and trehalose (both in the absence and presence of testosterone). In all cases, Bicalutamide and trehalose alone or in combination are all capable to reduce the number of ARpolyQ aggregates formed after testosterone treatment. All together, the data strongly suggest the presence of active autophagy capable to remove the cytoplasmically retained ARpolyQ.Figure 5.

Bottom Line: Thus, (i) prevention of ARpolyQ nuclear localization, combined with (ii) an increased ARpolyQ cytoplasmic clearance, should reduce its detrimental activity.Using the antiandrogen Bicalutamide (Casodex(®)), which slows down AR activation and nuclear translocation, and the disaccharide trehalose, an autophagy activator, we found that, in motoneurons, the two compounds together reduced ARpolyQ insoluble forms with higher efficiency than that obtained with single treatments.Collectively, these data suggest that the combinatory use of Bicalutamide and trehalose is a novel approach to facilitate ARpolyQ clearance that has to be tested in other cell types target of SBMA (i.e. muscle cells) and in vivo in animal models of SBMA.

View Article: PubMed Central - PubMed

Affiliation: Sezione di Biomedicina ed Endocrinologia, Dipartimento di Scienze Farmacologiche e Biomolecolari (DiSFeB), Centro di Eccellenza sulle Malattie Neurodegenerative, Università degli Studi di Milano, Milano 20133, Italy Centro InterUniversitario sulle Malattie Neurodegenerative, Università degli Studi di Firenze, Genova e Roma Tor Vergata, Milano 20133, Italy Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA and.

Show MeSH
Related in: MedlinePlus