PT  - JOURNAL ARTICLE
AU  - Emma V. Rusilowicz-Jones
AU  - Francesco G. Barone
AU  - Fernanda Martins Lopes
AU  - Elizabeth Stephen
AU  - Heather Mortiboys
AU  - Sylvie Urbé
AU  - Michael J. Clague
TI  - Benchmarking a highly selective USP30 inhibitor for enhancement of mitophagy and pexophagy
AID  - 10.1101/2021.04.28.441730
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.04.28.441730
4099  - http://biorxiv.org/content/early/2021/04/28/2021.04.28.441730.short
4100  - http://biorxiv.org/content/early/2021/04/28/2021.04.28.441730.full
AB  - The deubiquitylase USP30 is an actionable target considered for treatment of conditions associated with defects in the PINK1/Parkin pathway leading to mitophagy. These include Parkinson’s disease and pulmonary fibrosis. We provide a detailed cell biological characterisation of a benzenesulphonamide molecule, compound 39, that has previously been reported to inhibit USP30 in an in vitro enzymatic assay. The current compound offers increased selectivity over previously described inhibitors. It enhances mitophagy and generates a signature response for USP30 inhibition following mitochondrial depolarisation. This includes enhancement of TOM20 and SYNJ2BP ubiquitylation and phosphoubiquitin accumulation, alongside increased mitophagy. In dopaminergic neurons, generated from Parkinson’s disease patients carrying loss of function Parkin mutations, compound 39 could significantly restore mitophagy to a level approaching control values. USP30 is located on both mitochondria and peroxisomes and has also been linked to the PINK1 independent pexophagy pathway. Using a fluorescence reporter of pexophagy expressed in U20S cells, we observe increased pexophagy upon application of compound 39 that recapitulates the previously described effect for USP30 depletion. This provides the first pharmacological intervention with a synthetic molecule to enhance peroxisome turnover.Competing Interest StatementThe authors have declared no competing interest.