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Antiprion drugs 6-aminophenanthridine and guanabenz reduce PABPN1 toxicity and aggregation in oculopharyngeal muscular dystrophy.

Barbezier N, Chartier A, Bidet Y, Buttstedt A, Voisset C, Galons H, Blondel M, Schwarz E, Simonelig M - EMBO Mol Med (2011)

Bottom Line: We show that deletions of the ribosomal DNA locus reduce OPMD phenotypes and act synergistically with sub-effective doses of 6AP.In a complementary approach, we demonstrate that ribosomal RNA accelerates in vitro fibril formation of PABPN1 N-terminal domain.These results reveal the conserved role of ribosomal RNA in different protein aggregation disorders and identify 6AP and GA as general anti-aggregation molecules.

View Article: PubMed Central - PubMed

Affiliation: mRNA Regulation and Development, Institut de Génétique Humaine, CNRS UPR 1142, Montpellier Cedex 5, France.

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6AP reduces the wing position defects resulting from expression of alanine-expanded PABPN1 in musclesUAS-PABPN1-17ala/+; Mhc-Gal4/+ first instar larvae were raised at 18°C on instant Drosophila medium supplemented with increasing concentrations of 6AP, 6APi or DMSO alone. The day of birth (day 1), adult males were transferred onto fresh medium with the same concentration of drug and scored for wing position at day 6 (n = 120–320). Oral administration of 6AP reduces the percentage of wing position defects in a dose-dependent manner (left panel), whereas 6APi shows no effect (right panel).Western blot of thorax protein extracts from UAS-PABPN1-17ala/+; Mhc-Gal4/+ individuals at day 6, fed from larval stages with 400 µM of 6AP or DMSO alone. The blot was probed with anti-PABPN1, α-tubulin was used as a loading control.UAS-PABPN1-17ala/+; Mhc-Gal4/+ embryos were raised on regular Drosophila medium at 18°C. The day of birth (day 1), adult males were transferred onto fresh instant Drosophila medium supplemented with 6AP, 6APi or DMSO alone. Wing position defects were scored at day 6 (n = 40–120). Because of the shorter period of exposure to the drug and the reduced taking of food by adults compared to larvae, concentrations of 6AP higher than 400 µM were found to be non-toxic.UAS-PABPN1-17ala/+; Mhc-Gal4/+ adults were transferred to 6AP or DMSO alone-supplemented medium at day 2. Wing position defects were scored at days 4, 5 and 6 (n = 112–119).UAS-PABPN1-17ala/+; Mhc-Gal4/+ adults with wing position defects were selected at day 3 and transferred immediately to 6AP or DMSO alone-supplemented medium, wing position defects were scored at day 4 (n = 100). ***p-value <10−4, **p-value < 0.01 and *p-value < 0.05 using the χ2 test.
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fig02: 6AP reduces the wing position defects resulting from expression of alanine-expanded PABPN1 in musclesUAS-PABPN1-17ala/+; Mhc-Gal4/+ first instar larvae were raised at 18°C on instant Drosophila medium supplemented with increasing concentrations of 6AP, 6APi or DMSO alone. The day of birth (day 1), adult males were transferred onto fresh medium with the same concentration of drug and scored for wing position at day 6 (n = 120–320). Oral administration of 6AP reduces the percentage of wing position defects in a dose-dependent manner (left panel), whereas 6APi shows no effect (right panel).Western blot of thorax protein extracts from UAS-PABPN1-17ala/+; Mhc-Gal4/+ individuals at day 6, fed from larval stages with 400 µM of 6AP or DMSO alone. The blot was probed with anti-PABPN1, α-tubulin was used as a loading control.UAS-PABPN1-17ala/+; Mhc-Gal4/+ embryos were raised on regular Drosophila medium at 18°C. The day of birth (day 1), adult males were transferred onto fresh instant Drosophila medium supplemented with 6AP, 6APi or DMSO alone. Wing position defects were scored at day 6 (n = 40–120). Because of the shorter period of exposure to the drug and the reduced taking of food by adults compared to larvae, concentrations of 6AP higher than 400 µM were found to be non-toxic.UAS-PABPN1-17ala/+; Mhc-Gal4/+ adults were transferred to 6AP or DMSO alone-supplemented medium at day 2. Wing position defects were scored at days 4, 5 and 6 (n = 112–119).UAS-PABPN1-17ala/+; Mhc-Gal4/+ adults with wing position defects were selected at day 3 and transferred immediately to 6AP or DMSO alone-supplemented medium, wing position defects were scored at day 4 (n = 100). ***p-value <10−4, **p-value < 0.01 and *p-value < 0.05 using the χ2 test.

Mentions: Expression of PABPN1-17ala with Mhc-Gal4 leads to progressive defects in wing position (wings up or down). At 18°C, abnormal wing position appears at day 3 of adulthood with less than 20% of individuals showing defects, and this percentage reaches 90% at day 6 (Chartier et al, 2006). We analyzed the effect of feeding OPMD larvae and adults with increasing concentrations of 6AP, ranging from 250 to 400 µM. Wing position defects were scored at day 6 of adulthood, when 90% of individuals show defects in the absence of the drug (Fig 2A). A dose-dependent beneficial effect of 6AP was visible. The strongest effect was obtained in the presence of 400 µM 6AP, the percentage of individuals with wing position defects decreased to 50%. An identical experiment performed in the presence of the control molecule 6APi showed that this molecule had no effect (Fig 2A). We found that 400 µM of 6AP was still beneficial, although to a lesser extent, when provided during larval stages only, or from third instar larval to adult stages (Supplementary Fig 3). We verified that the positive effect of 6AP did not result from reduced levels of PABPN1-17ala in thoracic muscles of individuals fed with 6AP (Fig 2B).


Antiprion drugs 6-aminophenanthridine and guanabenz reduce PABPN1 toxicity and aggregation in oculopharyngeal muscular dystrophy.

Barbezier N, Chartier A, Bidet Y, Buttstedt A, Voisset C, Galons H, Blondel M, Schwarz E, Simonelig M - EMBO Mol Med (2011)

6AP reduces the wing position defects resulting from expression of alanine-expanded PABPN1 in musclesUAS-PABPN1-17ala/+; Mhc-Gal4/+ first instar larvae were raised at 18°C on instant Drosophila medium supplemented with increasing concentrations of 6AP, 6APi or DMSO alone. The day of birth (day 1), adult males were transferred onto fresh medium with the same concentration of drug and scored for wing position at day 6 (n = 120–320). Oral administration of 6AP reduces the percentage of wing position defects in a dose-dependent manner (left panel), whereas 6APi shows no effect (right panel).Western blot of thorax protein extracts from UAS-PABPN1-17ala/+; Mhc-Gal4/+ individuals at day 6, fed from larval stages with 400 µM of 6AP or DMSO alone. The blot was probed with anti-PABPN1, α-tubulin was used as a loading control.UAS-PABPN1-17ala/+; Mhc-Gal4/+ embryos were raised on regular Drosophila medium at 18°C. The day of birth (day 1), adult males were transferred onto fresh instant Drosophila medium supplemented with 6AP, 6APi or DMSO alone. Wing position defects were scored at day 6 (n = 40–120). Because of the shorter period of exposure to the drug and the reduced taking of food by adults compared to larvae, concentrations of 6AP higher than 400 µM were found to be non-toxic.UAS-PABPN1-17ala/+; Mhc-Gal4/+ adults were transferred to 6AP or DMSO alone-supplemented medium at day 2. Wing position defects were scored at days 4, 5 and 6 (n = 112–119).UAS-PABPN1-17ala/+; Mhc-Gal4/+ adults with wing position defects were selected at day 3 and transferred immediately to 6AP or DMSO alone-supplemented medium, wing position defects were scored at day 4 (n = 100). ***p-value <10−4, **p-value < 0.01 and *p-value < 0.05 using the χ2 test.
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Related In: Results  -  Collection

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fig02: 6AP reduces the wing position defects resulting from expression of alanine-expanded PABPN1 in musclesUAS-PABPN1-17ala/+; Mhc-Gal4/+ first instar larvae were raised at 18°C on instant Drosophila medium supplemented with increasing concentrations of 6AP, 6APi or DMSO alone. The day of birth (day 1), adult males were transferred onto fresh medium with the same concentration of drug and scored for wing position at day 6 (n = 120–320). Oral administration of 6AP reduces the percentage of wing position defects in a dose-dependent manner (left panel), whereas 6APi shows no effect (right panel).Western blot of thorax protein extracts from UAS-PABPN1-17ala/+; Mhc-Gal4/+ individuals at day 6, fed from larval stages with 400 µM of 6AP or DMSO alone. The blot was probed with anti-PABPN1, α-tubulin was used as a loading control.UAS-PABPN1-17ala/+; Mhc-Gal4/+ embryos were raised on regular Drosophila medium at 18°C. The day of birth (day 1), adult males were transferred onto fresh instant Drosophila medium supplemented with 6AP, 6APi or DMSO alone. Wing position defects were scored at day 6 (n = 40–120). Because of the shorter period of exposure to the drug and the reduced taking of food by adults compared to larvae, concentrations of 6AP higher than 400 µM were found to be non-toxic.UAS-PABPN1-17ala/+; Mhc-Gal4/+ adults were transferred to 6AP or DMSO alone-supplemented medium at day 2. Wing position defects were scored at days 4, 5 and 6 (n = 112–119).UAS-PABPN1-17ala/+; Mhc-Gal4/+ adults with wing position defects were selected at day 3 and transferred immediately to 6AP or DMSO alone-supplemented medium, wing position defects were scored at day 4 (n = 100). ***p-value <10−4, **p-value < 0.01 and *p-value < 0.05 using the χ2 test.
Mentions: Expression of PABPN1-17ala with Mhc-Gal4 leads to progressive defects in wing position (wings up or down). At 18°C, abnormal wing position appears at day 3 of adulthood with less than 20% of individuals showing defects, and this percentage reaches 90% at day 6 (Chartier et al, 2006). We analyzed the effect of feeding OPMD larvae and adults with increasing concentrations of 6AP, ranging from 250 to 400 µM. Wing position defects were scored at day 6 of adulthood, when 90% of individuals show defects in the absence of the drug (Fig 2A). A dose-dependent beneficial effect of 6AP was visible. The strongest effect was obtained in the presence of 400 µM 6AP, the percentage of individuals with wing position defects decreased to 50%. An identical experiment performed in the presence of the control molecule 6APi showed that this molecule had no effect (Fig 2A). We found that 400 µM of 6AP was still beneficial, although to a lesser extent, when provided during larval stages only, or from third instar larval to adult stages (Supplementary Fig 3). We verified that the positive effect of 6AP did not result from reduced levels of PABPN1-17ala in thoracic muscles of individuals fed with 6AP (Fig 2B).

Bottom Line: We show that deletions of the ribosomal DNA locus reduce OPMD phenotypes and act synergistically with sub-effective doses of 6AP.In a complementary approach, we demonstrate that ribosomal RNA accelerates in vitro fibril formation of PABPN1 N-terminal domain.These results reveal the conserved role of ribosomal RNA in different protein aggregation disorders and identify 6AP and GA as general anti-aggregation molecules.

View Article: PubMed Central - PubMed

Affiliation: mRNA Regulation and Development, Institut de Génétique Humaine, CNRS UPR 1142, Montpellier Cedex 5, France.

Show MeSH
Related in: MedlinePlus