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Induction of autophagy by a novel small molecule improves aβ pathology and ameliorates cognitive deficits.

Chu C, Zhang X, Ma W, Li L, Wang W, Shang L, Fu P - PLoS ONE (2013)

Bottom Line: GTM-1 modulates autophagy in an Akt-independent and mTOR-independent manner.In addition, we demonstrated that GTM-1 enhances autophagy clearance and reverses the downregulation of autophagy flux by thapsigargin and asparagine.Furthermore, administration of GTM-1 attenuated Aβ pathology and ameliorated cognitive deficits in AD mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, The First People's Hospital of Yangzhou, Jiang Su, PR China.

ABSTRACT
Growing evidence has demonstrated a neuroprotective role of autophagy in Alzheimer's disease (AD). Thus, autophagy has been regarded as a potential therapeutic target, attracting increasing interest in pharmaceutical autophagy modulation by small molecules. We designed a two-cycle screening strategy on the basis of imaging high-throughout screening (HTS) and cellular toxicity assay, and have identified a novel autophagy inducer known as GTM-1. We further showed that GTM-1 exhibits dual activities, such as autophagy induction and antagonism against Aβ-oligomer toxicity. GTM-1 modulates autophagy in an Akt-independent and mTOR-independent manner. In addition, we demonstrated that GTM-1 enhances autophagy clearance and reverses the downregulation of autophagy flux by thapsigargin and asparagine. Furthermore, administration of GTM-1 attenuated Aβ pathology and ameliorated cognitive deficits in AD mice.

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GTM-1 up-regulates autophagy level.A, SH-SY5Y/LC3-GFP and MC65 cells were treated with 20 µM GTM-1 for 12 hrs. Next, the cell lysates were analyzed using western blotting with anti-LC3 and anti-β-tubulin (as a control). B, Left: SH-SY5Y and MC65 cells were treated with 20 µM GTM-1 for 6 hrs. Next, the cell lysates were analyzed using western blotting with anti-LC3 and anti-β-tubulin (as a control). Right: quantification of the Western blot images of the ratio between LC3-II and tubulin. C, MC65 cells were treated with vehicle control (1% DMSO) or GTM-1 (20 µM) for 8 hrs. The cells were then fixed with glutaraldehyde and prepared for EM analysis. Bar, 1∶11,000. Arrows indicate double and multi-membrane autophagosomic vesicles.
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pone-0065367-g002: GTM-1 up-regulates autophagy level.A, SH-SY5Y/LC3-GFP and MC65 cells were treated with 20 µM GTM-1 for 12 hrs. Next, the cell lysates were analyzed using western blotting with anti-LC3 and anti-β-tubulin (as a control). B, Left: SH-SY5Y and MC65 cells were treated with 20 µM GTM-1 for 6 hrs. Next, the cell lysates were analyzed using western blotting with anti-LC3 and anti-β-tubulin (as a control). Right: quantification of the Western blot images of the ratio between LC3-II and tubulin. C, MC65 cells were treated with vehicle control (1% DMSO) or GTM-1 (20 µM) for 8 hrs. The cells were then fixed with glutaraldehyde and prepared for EM analysis. Bar, 1∶11,000. Arrows indicate double and multi-membrane autophagosomic vesicles.

Mentions: To confirm the autophagy stimulation of GTM-1, SH-SY5Y and MC65 cells treated with GTM-1 were assayed by LC3 western blotting. Treatment with GTM-1 showed a robust LC3-II band (Fig. 2A), which was consistent with the HCS assay results of LC3-GFP puncta (Fig. 1B, 1C, 1D). Further studies performed in SH-SY5Y cells showed that GTM-1 can stimulate autophagy in the presence of Aβ42 (Fig. 2B).


Induction of autophagy by a novel small molecule improves aβ pathology and ameliorates cognitive deficits.

Chu C, Zhang X, Ma W, Li L, Wang W, Shang L, Fu P - PLoS ONE (2013)

GTM-1 up-regulates autophagy level.A, SH-SY5Y/LC3-GFP and MC65 cells were treated with 20 µM GTM-1 for 12 hrs. Next, the cell lysates were analyzed using western blotting with anti-LC3 and anti-β-tubulin (as a control). B, Left: SH-SY5Y and MC65 cells were treated with 20 µM GTM-1 for 6 hrs. Next, the cell lysates were analyzed using western blotting with anti-LC3 and anti-β-tubulin (as a control). Right: quantification of the Western blot images of the ratio between LC3-II and tubulin. C, MC65 cells were treated with vehicle control (1% DMSO) or GTM-1 (20 µM) for 8 hrs. The cells were then fixed with glutaraldehyde and prepared for EM analysis. Bar, 1∶11,000. Arrows indicate double and multi-membrane autophagosomic vesicles.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3672196&req=5

pone-0065367-g002: GTM-1 up-regulates autophagy level.A, SH-SY5Y/LC3-GFP and MC65 cells were treated with 20 µM GTM-1 for 12 hrs. Next, the cell lysates were analyzed using western blotting with anti-LC3 and anti-β-tubulin (as a control). B, Left: SH-SY5Y and MC65 cells were treated with 20 µM GTM-1 for 6 hrs. Next, the cell lysates were analyzed using western blotting with anti-LC3 and anti-β-tubulin (as a control). Right: quantification of the Western blot images of the ratio between LC3-II and tubulin. C, MC65 cells were treated with vehicle control (1% DMSO) or GTM-1 (20 µM) for 8 hrs. The cells were then fixed with glutaraldehyde and prepared for EM analysis. Bar, 1∶11,000. Arrows indicate double and multi-membrane autophagosomic vesicles.
Mentions: To confirm the autophagy stimulation of GTM-1, SH-SY5Y and MC65 cells treated with GTM-1 were assayed by LC3 western blotting. Treatment with GTM-1 showed a robust LC3-II band (Fig. 2A), which was consistent with the HCS assay results of LC3-GFP puncta (Fig. 1B, 1C, 1D). Further studies performed in SH-SY5Y cells showed that GTM-1 can stimulate autophagy in the presence of Aβ42 (Fig. 2B).

Bottom Line: GTM-1 modulates autophagy in an Akt-independent and mTOR-independent manner.In addition, we demonstrated that GTM-1 enhances autophagy clearance and reverses the downregulation of autophagy flux by thapsigargin and asparagine.Furthermore, administration of GTM-1 attenuated Aβ pathology and ameliorated cognitive deficits in AD mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, The First People's Hospital of Yangzhou, Jiang Su, PR China.

ABSTRACT
Growing evidence has demonstrated a neuroprotective role of autophagy in Alzheimer's disease (AD). Thus, autophagy has been regarded as a potential therapeutic target, attracting increasing interest in pharmaceutical autophagy modulation by small molecules. We designed a two-cycle screening strategy on the basis of imaging high-throughout screening (HTS) and cellular toxicity assay, and have identified a novel autophagy inducer known as GTM-1. We further showed that GTM-1 exhibits dual activities, such as autophagy induction and antagonism against Aβ-oligomer toxicity. GTM-1 modulates autophagy in an Akt-independent and mTOR-independent manner. In addition, we demonstrated that GTM-1 enhances autophagy clearance and reverses the downregulation of autophagy flux by thapsigargin and asparagine. Furthermore, administration of GTM-1 attenuated Aβ pathology and ameliorated cognitive deficits in AD mice.

Show MeSH
Related in: MedlinePlus