2. Academic Validation
  3. Genome-wide CRISPR screen reveals v-ATPase as a drug target to lower levels of ALS protein ataxin-2

Genome-wide CRISPR screen reveals v-ATPase as a drug target to lower levels of ALS protein ataxin-2

  • Cell Rep. 2022 Oct 25;41(4):111508. doi: 10.1016/j.celrep.2022.111508.
Garam Kim 1,  Lisa Nakayama 2,  Jacob A Blum 1,  Tetsuya Akiyama 2,  Steven Boeynaems 2,  Meenakshi Chakraborty 2,  Julien Couthouis 2,  Eduardo Tassoni-Tsuchida 3,  Caitlin M Rodriguez 2,  Michael C Bassik 2,  Aaron D Gitler 4
Affiliations

Affiliations

  • 1 Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA; Stanford Neurosciences Interdepartmental Program, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 2 Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 3 Department of Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 4 Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA. Electronic address: agitler@stanford.edu.
PMID: 36288714 DOI: 10.1016/j.celrep.2022.111508
Abstract

Mutations in the ataxin-2 gene (ATXN2) cause the neurodegenerative disorders amyotrophic lateral sclerosis (ALS) and spinocerebellar ataxia type 2 (SCA2). A therapeutic strategy using Antisense Oligonucleotides targeting ATXN2 has entered clinical trial in humans. Additional ways to decrease ataxin-2 levels could lead to cheaper or less invasive therapies and elucidate how ataxin-2 is normally regulated. Here, we perform a genome-wide fluorescence-activated cell sorting (FACS)-based CRISPR-Cas9 screen in human cells and identify genes encoding components of the lysosomal vacuolar ATPase (v-ATPase) as modifiers of endogenous ataxin-2 protein levels. Multiple FDA-approved small molecule v-ATPase inhibitors lower ataxin-2 protein levels in mouse and human neurons, and oral administration of at least one of these drugs-etidronate-is sufficient to decrease ataxin-2 in the brains of mice. Together, we propose v-ATPase as a drug target for ALS and SCA2 and demonstrate the value of FACS-based screens in identifying genetic-and potentially druggable-modifiers of human disease proteins.

Keywords

ALS; CP: Neuroscience; FACS; SCA2; TDP-43; ataxin-2; bisphosphonate; etidronate; genetic screens; small-molecule therapy; v-ATPase.

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