Inhibition of RNA splicing triggers CHMP7 nuclear entry, impacting TDP-43 function and leading to the onset of ALS cellular phenotypes
Amyotrophic lateral sclerosis (ALS) is linked to the reduction of certain nucleoporins in neurons. Increased nuclear localization of charged multivesicular body protein 7 (CHMP7), a protein involved in nuclear pore surveillance, has been identified as a key factor damaging nuclear pores and disrupting transport. Using CRISPR-based microRaft, followed by gRNA identification (CRaft-ID), we discovered 55 RNA-binding proteins (RBPs) that influence CHMP7 localization, including SmD1, a survival of motor neuron (SMN) complex component. Immunoprecipitation-mass spectrometry (IP-MS) and enhanced crosslinking and immunoprecipitation (CLIP) analyses revealed CHMP7's interactions with SmD1, small nuclear RNAs, and splicing factor mRNAs in motor neurons (MNs). ALS induced pluripotent stem cell (iPSC)-MNs show reduced SmD1 expression, and inhibiting SmD1/SMN complex increased CHMP7 nuclear localization. Crucially, overexpressing SmD1 in ALS iPSC-MNs restored CHMP7's cytoplasmic localization and corrected STMN2 splicing. Our findings suggest that early ALS pathogenesis is driven by SMN complex dysregulation.
Authors
- Anthony Q. Vu (ORCID: https://orcid.org/0000-0001-8922-6409)
- Cara R. Schiavon (ORCID: https://orcid.org/0000-0002-9311-2145)
- Lena Annika Street (ORCID: https://orcid.org/0000-0002-5473-6713)
- Alyssa N. Coyne (ORCID: https://orcid.org/0000-0002-3658-5325)
- Steven M. Blue (ORCID: https://orcid.org/0000-0003-0279-6690)
- Elena Rebollo (ORCID: https://orcid.org/0000-0001-8899-0317)
- Jeffrey D. Rothstein (ORCID: https://orcid.org/0000-0003-2001-8470)
- Jasmine R. Mueller (ORCID: https://orcid.org/0000-0001-6236-8001)
- Assael A. Madrigal (ORCID: https://orcid.org/0000-0002-3311-5256)
- Dylan C. Lam (ORCID: https://orcid.org/0000-0001-6472-7127)
- Wenhao Jin (ORCID: https://orcid.org/0000-0002-7885-3527)
- Norah Al-Azzam (ORCID: https://orcid.org/0000-0002-8960-8236)
- Uri Manor (ORCID: https://orcid.org/0000-0002-9802-1955)
- Marko Jovanović (ORCID: https://orcid.org/0000-0001-9707-3377)
- Orel Mizrahi (ORCID: https://orcid.org/0000-0003-2189-5731)
- Chloe Nguyen
- G Yeo
- Jack T. Naritomi (ORCID: https://orcid.org/0000-0001-7746-7467)
- Benjamin Lee (ORCID: https://orcid.org/0000-0002-1171-4741)
- W. L. Ford
- Vaishali Gautam
- Anthony Avina
- Yashwin L Madakamutil
- Jenny H To
Institutions
- Salk Institute for Biological Studies (US)
- Johns Hopkins University (US)
- Johns Hopkins Medicine (US)
- University of California San Diego (US)
- Torrey Pines Institute For Molecular Studies (US)
- Innovative Research (United States) (US)
- Columbia University (US)
Publication Details
- Journal
- Neuron
- Published
- 2024-10-31
- DOI
- https://doi.org/10.1016/j.neuron.2024.10.007
- Citations
- 7
- Primary Topic
- Amyotrophic Lateral Sclerosis Research
- Type
- article
- Field-Weighted Citation Impact
- 2.58
Funders
- National Science Foundation
- Silicon Valley Community Foundation
- Waitt Foundation
- National University of Singapore
- National Institutes of Health
- National Cancer Institute