Nanoscopic investigation of C9orf72 poly-GA oligomers on nuclear membrane disruption by a photoinducible platform

Hung-Ming Chien; Ruei-Yu He; Chi-Chang Lee; Yung-An Huang; I-Ju Hung; Kai-Ting Hou; Jye-Chian Hsiao; Po-Chao Lu; Diksha Agnihotri; Eric Hwang; Joseph Jen-Tse Huang

doi:10.1038/s42004-021-00547-6

Nanoscopic investigation of C9orf72 poly-GA oligomers on nuclear membrane disruption by a photoinducible platform

Commun Chem. 2021 Jul 23;4(1):111. doi: 10.1038/s42004-021-00547-6.

Authors

Hung-Ming Chien^{1

2

3}, Ruei-Yu He¹, Chi-Chang Lee¹, Yung-An Huang^{1

4}, I-Ju Hung^{1

5}, Kai-Ting Hou¹, Jye-Chian Hsiao¹, Po-Chao Lu^{1

3

6}, Diksha Agnihotri^{1

7}, Eric Hwang^{4

8

9

10}, Joseph Jen-Tse Huang^{11

12

13

14}

Affiliations

¹ Institute of Chemistry, Academia Sinica, Taipei City, Taiwan.
² Department of Chemistry, National Taiwan University, Taipei City, Taiwan.
³ Chemical Biology and Molecular Biophysics, Taiwan International Graduate Program, Academia Sinica and National Taiwan University, Taipei City, Taiwan.
⁴ Department of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsinchu City, Taiwan.
⁵ Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei City, Taiwan.
⁶ Department and Graduate Institute of Pharmacology, National Taiwan University, Taipei City, Taiwan.
⁷ Taiwan International Graduate Program in Interdisciplinary Neuroscience, National Taiwan University and Academia Sinica, Taipei City, Taiwan.
⁸ Institute of Molecular Medicine and Bioengineering, National Yang Ming Chiao Tung University, Hsinchu City, Taiwan.
⁹ Institute of Bioinformatics and Systems Biology, National Yang Ming Chiao Tung University, Hsinchu City, Taiwan.
¹⁰ Center for Intelligent Drug Systems and Smart Bio-devices (IDS2B), National Yang Ming Chiao Tung University, Hsinchu City, Taiwan.
¹¹ Institute of Chemistry, Academia Sinica, Taipei City, Taiwan. jthuang@gate.sinica.edu.tw.
¹² Chemical Biology and Molecular Biophysics, Taiwan International Graduate Program, Academia Sinica and National Taiwan University, Taipei City, Taiwan. jthuang@gate.sinica.edu.tw.
¹³ Department of Applied Chemistry, National Chiayi University, Chiayi City, Taiwan. jthuang@gate.sinica.edu.tw.
¹⁴ Neuroscience Program of Academia Sinica, Academia Sinica, Taipei, Taiwan. jthuang@gate.sinica.edu.tw.

Abstract

Glycine-alanine dipeptide repeats (GA DPRs) translated from the mutated C9orf72 gene have recently been correlated with amyotrophic lateral sclerosis (ALS). While GA DPRs aggregates have been suggested as amyloid, the biophysical features and cytotoxicity of GA DPRs oligomers has not been explored due to its unstable nature. In this study, we develop a photoinducible platform based on methoxynitrobenzene chemistry to enrich GA DPRs that allows monitoring the oligomerization process of GA DPRs in cells. By applying advanced microscopies, we examined the GA DPRs oligomerization process nanoscopically in a time-dependent manner. We provided direct evidences to demonstrate GA DPRs oligomers rather than nanofibrils disrupt nuclear membrane. Moreover, we found GA DPRs hamper nucleocytoplasmic transport in cells and cause cytosolic retention of TAR DNA-binding protein 43 in cortical neurons. Our results highlight the toxicity of GA DPRs oligomers, which is a key step toward elucidating the pathological roles of C9orf72 DPRs.