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The contribution of proteasomal impairment to autophagy activation by C9orf72 poly-GA aggregates

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Abstract

Background

Poly-GA, a dipeptide repeat protein unconventionally translated from GGGGCC (G4C2) repeat expansions in C9orf72, is abundant in C9orf72-related amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) (C9orf72-ALS/FTD). Although the poly-GA aggregates have been identified in C9orf72-ALS/FTD neurons, the effects on UPS (ubiquitin–proteasome system) and autophagy and their exact molecular mechanisms have not been fully elucidated.

Results

Herein, our in vivo experiments indicate that the mice expressing ploy-GA with 150 repeats instead of 30 repeats exhibit significant aggregates in cells. Mice expressing 150 repeats ploy-GA shows behavioral deficits and activates autophagy in the brain. In vitro findings suggest that the poly-GA aggregates influence proteasomal by directly binding proteasome subunit PSMD2. Subsequently, the poly-GA aggregates activate phosphorylation and ubiquitination of p62 to recruit autophagosomes. Ultimately, the poly-GA aggregates lead to compensatory activation of autophagy. In vivo studies further reveal that rapamycin (autophagy activator) treatment significantly improves the degenerative symptoms and alleviates neuronal injury in mice expressing 150 repeats poly-GA. Meanwhile, rapamycin administration to mice expressing 150 repeats poly-GA reduces neuroinflammation and aggregates in the brain.

Conclusion

In summary, we elucidate the relationship between poly-GA in the proteasome and autophagy: when poly-GA forms complexes with the proteasome, it recruits autophagosomes and affects proteasome function. Our study provides support for further promoting the comprehension of the pathogenesis of C9orf72, which may bring a hint for the exploration of rapamycin for the treatment of ALS/FTD.

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Availability of data and materials

All data generated or analyzed during this study are included in this published article and its supplementary information files. Further data supporting the findings of this study are available from the corresponding authors on request.

Abbreviations

ALS:

Amyotrophic lateral sclerosis

FTD:

Frontotemporal dementia

C9orf72:

Chromosome 9 open reading frame 72

poly-GA:

Poly-Glycine Alanine

GFAP:

Glial fibrillary acidic protein

Iba1:

Ionized calcium-binding adapter molecule 1

UPS:

Ubiquitin − proteasome system

DPRs:

Dipeptide repeat proteins

PolyQ:

Polyglutamine

HD:

Huntington’s disease

AD:

Alzheimer’s disease

LIR:

LC3-interacting region

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Acknowledgements

We would like to thank Dr. Ruimin Huang (Shanghai Institute of Materia Medica) for his critical comments and suggestions on our manuscript; Thank Dr. Kai Fu (Xiangya Hospital Center South University) for his advice and critical reading of this manuscript. Thank Dr. Zhaobing Gao (Shanghai Institute of Materia Medica) for helpful comments.

Funding

This work was supported by the Foundation of Shanghai Science and Technology Committee (No. 21ZR1475100, 21DZ2291100), the Fundamental Research Funds for the State Key Laboratory of Drug Research (SIMM2103ZZ-01), the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine (No: ZYYCXTD-D-202210).

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Authors and Affiliations

  1. Center for Drug Safety Evaluation and Research, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai, China

    Mei Pu, Yusi Tai, Huijie Guo, Jing Chen, Xinming Qi, Zhouteng Tao & Jin Ren

  2. School of Life Science and Technology, ShanghaiTech University, Shanghai, China

    Mei Pu, Huijie Guo & Jin Ren

  3. University of Chinese Academy of Sciences, Beijing, China

    Mei Pu, Yusi Tai, Huijie Guo & Jin Ren

  4. School of Chinese Materia Medica, Nanjing University of Chinese Medicine, Nanjing, China

    Luyang Yuan & Jin Ren

  5. Key Laboratory of Receptor Research, Department of Neuropharmacology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China

    Yu Zhang

  6. Laboratory of Molecular Neuropathology, Jiangsu Key Laboratory of Neuropsychiatric Diseases and Department of Pharmacology, College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, China

    Zongbing Hao & Guanghui Wang

  7. State Key Laboratory of Transient Optics and Photonics, Xi’an Institute of Optics and Precision Mechanics, Chinese Academy of Sciences, Xi’an, China

    Zhouteng Tao

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  1. Mei Pu
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  2. Yusi Tai
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  3. Luyang Yuan
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  4. Yu Zhang
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  11. Jin Ren
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Contributions

All the authors contributed significantly to the study. MP, ZT: conceptualization. MP, YT, HG, LY, YZ, ZH: methodology. MP: software. MP, ZT: writing—original draft. GW, ZT, JR: writing—reviewing and editing. ZT, JC, XQ, JR: supervision, funding acquisition. All authors have read and agreed to the published version of the manuscript.

Corresponding authors

Correspondence to Guanghui Wang, Zhouteng Tao or Jin Ren.

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No human data or tissue were involved. All animal procedures received approval from the Animal Care and Use Committee of the Shanghai Institute of Materia Medica, Chinese Academy of Sciences (Shanghai, China).

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Pu, M., Tai, Y., Yuan, L. et al. The contribution of proteasomal impairment to autophagy activation by C9orf72 poly-GA aggregates. Cell. Mol. Life Sci. 79, 501 (2022). https://doi.org/10.1007/s00018-022-04518-5

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