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Indole and synthetic derivative activate chaperone expression to reduce polyQ aggregation in SCA17 neuronal cell and slice culture models.

Kung PJ, Tao YC, Hsu HC, Chen WL, Lin TH, Janreddy D, Yao CF, Chang KH, Lin JY, Su MT, Wu CH, Lee-Chen GJ, Hsieh-Li HM - Drug Des Devel Ther (2014)

Bottom Line: The molecular chaperones act in preventing protein aggregation to ameliorate downstream harmful events.We found that indole and NC001-8 up-regulated chaperone expression to reduce polyQ aggregation in neuronal differentiated TBP/Q79 cells.Our results demonstrate how indole and derivative NC001-8 reduce polyQ aggregation to support their therapeutic potentials in SCA17 treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Science, National Taiwan Normal University, Taipei, Taiwan.

ABSTRACT
In spinocerebellar ataxia type 17 (SCA17), the expansion of a translated CAG repeat in the TATA box binding protein (TBP) gene results in a long polyglutamine (polyQ) tract in the TBP protein, leading to intracellular accumulation of aggregated TBP and cell death. The molecular chaperones act in preventing protein aggregation to ameliorate downstream harmful events. In this study, we used Tet-On SH-SY5Y cells with inducible SCA17 TBP/Q79-green fluorescent protein (GFP) expression to test indole and synthetic derivative NC001-8 for neuroprotection. We found that indole and NC001-8 up-regulated chaperone expression to reduce polyQ aggregation in neuronal differentiated TBP/Q79 cells. The effects on promoting neurite outgrowth and on reduction of aggregation on Purkinje cells were also confirmed with cerebellar primary and slice cultures of SCA17 transgenic mice. Our results demonstrate how indole and derivative NC001-8 reduce polyQ aggregation to support their therapeutic potentials in SCA17 treatment.

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Enhancement of HSF1 and chaperone expression and reduction of aggregation by indole and NC001-8 in neuronal SH-SY5Y TBP/Q79 cells.Notes: (A) Cells were pretreated with GGA, indole, or NC001-8 (100 nM) for 8 hours and TBP/Q79-GFP expression induced for 6 days. Relative HSF1, HSPA8, and HSPA1A expressions were analyzed by immunoblot analysis, using β-actin as a loading control (n=3). P-values were evaluated by one-way analysis of variance with post hoc LSD test. (B) Cells were treated with GGA, indole, or NC001-8 (100 nM) for 7 days, and relative aggregation assessed by HCA system (n=3). To normalize, the relative aggregation level in untreated cells is set as 100%.Abbreviations: Dox, doxycycline; GGA, geranylgeranylacetone; HSF1, heat shock transcription factor 1; HSPA8, heat shock 70 kDa protein 8; HSPA1A, heat shock 70 kDa protein 1A; Rel., relative; GFP, green fluorescent protein; HCA: high content analysis; 001-8, NC001-8; LSD, Fisher’s least significant difference.
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f4-dddt-8-1929: Enhancement of HSF1 and chaperone expression and reduction of aggregation by indole and NC001-8 in neuronal SH-SY5Y TBP/Q79 cells.Notes: (A) Cells were pretreated with GGA, indole, or NC001-8 (100 nM) for 8 hours and TBP/Q79-GFP expression induced for 6 days. Relative HSF1, HSPA8, and HSPA1A expressions were analyzed by immunoblot analysis, using β-actin as a loading control (n=3). P-values were evaluated by one-way analysis of variance with post hoc LSD test. (B) Cells were treated with GGA, indole, or NC001-8 (100 nM) for 7 days, and relative aggregation assessed by HCA system (n=3). To normalize, the relative aggregation level in untreated cells is set as 100%.Abbreviations: Dox, doxycycline; GGA, geranylgeranylacetone; HSF1, heat shock transcription factor 1; HSPA8, heat shock 70 kDa protein 8; HSPA1A, heat shock 70 kDa protein 1A; Rel., relative; GFP, green fluorescent protein; HCA: high content analysis; 001-8, NC001-8; LSD, Fisher’s least significant difference.

Mentions: The Flp-In SH-SY5Y TBP/Q79-GFP cells discussed earlier were used to examine whether indole and NC001-8 up-regulate HSF1, HSPA8, and HSPA1A expression to reduce aggregation. As shown in Figure 4A, induced expression of TBP/Q79 for 6 days attenuated the expression of HSF1 (72%; P=0.005), HSPA8 (78%; P=0.046), and HSPA1A (84%; P=0.047) compared with uninduced cells (100%). This reduction can be rescued by the addition of GGA, indole, or NC001-8 (100 nM), with significantly increased HSF1 (96%~111%; P=0.039~0.004), HSPA8 (99%~105%; P=0.024~0.011), and HSPA1A (106%~117%; P=0.008~0.001) expression compared with untreated cells (72%~84%). The treatment of GGA, indole, and NC001-8 led to 17% (P=0.001), 15% (P=0.002), and 14% (P=0.010), respectively, aggregation reduction in TBP/Q79 expressed differentiated neuronal cells (Figure 4B). These findings indicate that indole and NC001-8 up-regulated HSF1 and HSP70 chaperone expression to reduce TBP/Q79 aggregation in differentiated neuronal cell models.


Indole and synthetic derivative activate chaperone expression to reduce polyQ aggregation in SCA17 neuronal cell and slice culture models.

Kung PJ, Tao YC, Hsu HC, Chen WL, Lin TH, Janreddy D, Yao CF, Chang KH, Lin JY, Su MT, Wu CH, Lee-Chen GJ, Hsieh-Li HM - Drug Des Devel Ther (2014)

Enhancement of HSF1 and chaperone expression and reduction of aggregation by indole and NC001-8 in neuronal SH-SY5Y TBP/Q79 cells.Notes: (A) Cells were pretreated with GGA, indole, or NC001-8 (100 nM) for 8 hours and TBP/Q79-GFP expression induced for 6 days. Relative HSF1, HSPA8, and HSPA1A expressions were analyzed by immunoblot analysis, using β-actin as a loading control (n=3). P-values were evaluated by one-way analysis of variance with post hoc LSD test. (B) Cells were treated with GGA, indole, or NC001-8 (100 nM) for 7 days, and relative aggregation assessed by HCA system (n=3). To normalize, the relative aggregation level in untreated cells is set as 100%.Abbreviations: Dox, doxycycline; GGA, geranylgeranylacetone; HSF1, heat shock transcription factor 1; HSPA8, heat shock 70 kDa protein 8; HSPA1A, heat shock 70 kDa protein 1A; Rel., relative; GFP, green fluorescent protein; HCA: high content analysis; 001-8, NC001-8; LSD, Fisher’s least significant difference.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4206201&req=5

f4-dddt-8-1929: Enhancement of HSF1 and chaperone expression and reduction of aggregation by indole and NC001-8 in neuronal SH-SY5Y TBP/Q79 cells.Notes: (A) Cells were pretreated with GGA, indole, or NC001-8 (100 nM) for 8 hours and TBP/Q79-GFP expression induced for 6 days. Relative HSF1, HSPA8, and HSPA1A expressions were analyzed by immunoblot analysis, using β-actin as a loading control (n=3). P-values were evaluated by one-way analysis of variance with post hoc LSD test. (B) Cells were treated with GGA, indole, or NC001-8 (100 nM) for 7 days, and relative aggregation assessed by HCA system (n=3). To normalize, the relative aggregation level in untreated cells is set as 100%.Abbreviations: Dox, doxycycline; GGA, geranylgeranylacetone; HSF1, heat shock transcription factor 1; HSPA8, heat shock 70 kDa protein 8; HSPA1A, heat shock 70 kDa protein 1A; Rel., relative; GFP, green fluorescent protein; HCA: high content analysis; 001-8, NC001-8; LSD, Fisher’s least significant difference.
Mentions: The Flp-In SH-SY5Y TBP/Q79-GFP cells discussed earlier were used to examine whether indole and NC001-8 up-regulate HSF1, HSPA8, and HSPA1A expression to reduce aggregation. As shown in Figure 4A, induced expression of TBP/Q79 for 6 days attenuated the expression of HSF1 (72%; P=0.005), HSPA8 (78%; P=0.046), and HSPA1A (84%; P=0.047) compared with uninduced cells (100%). This reduction can be rescued by the addition of GGA, indole, or NC001-8 (100 nM), with significantly increased HSF1 (96%~111%; P=0.039~0.004), HSPA8 (99%~105%; P=0.024~0.011), and HSPA1A (106%~117%; P=0.008~0.001) expression compared with untreated cells (72%~84%). The treatment of GGA, indole, and NC001-8 led to 17% (P=0.001), 15% (P=0.002), and 14% (P=0.010), respectively, aggregation reduction in TBP/Q79 expressed differentiated neuronal cells (Figure 4B). These findings indicate that indole and NC001-8 up-regulated HSF1 and HSP70 chaperone expression to reduce TBP/Q79 aggregation in differentiated neuronal cell models.

Bottom Line: The molecular chaperones act in preventing protein aggregation to ameliorate downstream harmful events.We found that indole and NC001-8 up-regulated chaperone expression to reduce polyQ aggregation in neuronal differentiated TBP/Q79 cells.Our results demonstrate how indole and derivative NC001-8 reduce polyQ aggregation to support their therapeutic potentials in SCA17 treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Science, National Taiwan Normal University, Taipei, Taiwan.

ABSTRACT
In spinocerebellar ataxia type 17 (SCA17), the expansion of a translated CAG repeat in the TATA box binding protein (TBP) gene results in a long polyglutamine (polyQ) tract in the TBP protein, leading to intracellular accumulation of aggregated TBP and cell death. The molecular chaperones act in preventing protein aggregation to ameliorate downstream harmful events. In this study, we used Tet-On SH-SY5Y cells with inducible SCA17 TBP/Q79-green fluorescent protein (GFP) expression to test indole and synthetic derivative NC001-8 for neuroprotection. We found that indole and NC001-8 up-regulated chaperone expression to reduce polyQ aggregation in neuronal differentiated TBP/Q79 cells. The effects on promoting neurite outgrowth and on reduction of aggregation on Purkinje cells were also confirmed with cerebellar primary and slice cultures of SCA17 transgenic mice. Our results demonstrate how indole and derivative NC001-8 reduce polyQ aggregation to support their therapeutic potentials in SCA17 treatment.

Show MeSH
Related in: MedlinePlus