Lysosomal proteomics links disturbances in lipid homeostasis and sphingolipid metabolism to CLN5 disease.

ABSTRACT: The CLN5 disease (MIM: 256731) represents a rare late-infantile form of neuronal ceroid lipofuscinosis (NCL) caused by mutations in the CLN5 gene that encodes a CLN5 protein (CLN5p), whose physiological roles remain unanswered. No cure is currently available for CLN5 patients and the opportunities for therapies are lagging behind. The role of lysosomes in neuro-pathophysiology of CLN5 disease represents an important topic since lysosomal proteins are directly involved in the primary mechanisms of neuronal injury occurring in various NCL forms. We developed and implemented a lysosome-focused, label-free quantitative proteomics approach followed by functional validations in both CLN5-knockout neuronal-like cell lines and Cln5-/- mice, to unravel affected pathways and modifying factors involved in this disease -scenario. Our results revealed a key role of CLN5p in lipid homeostasis and sphingolipid metabolism, representing a possible connection to the activation of mitochondria-driven cell death and mitophagy, other features of CLN5 disease. To translate findings from preclinical models to patients, we evaluated in vitro if FDA-approved drugs promoting autophagy via TFEB activation or inhibition of the glucosylceramide synthase could modulate ROS and lipids overproduction. We further tested in vivo, the efficacy of drugs in rescuing the locomotor function in a newly generated cln5 knockdown zebrafish model, recapitulating most of the pathological features seen in NCL. In summary, our data advances general understanding of pathogenetic mechanisms in CLN5 disease, stipulating new pharmacological treatments.

INSTRUMENT(S): Synapt G2-S HDMS

ORGANISM(S): Homo Sapiens (ncbitaxon:9606) Mus Musculus (ncbitaxon:10090)

SUBMITTER: Maciej Lalowski   Filippo M. Santorelli  

PROVIDER: MSV000088517 | MassIVE | Sun Dec 05 06:51:00 GMT 2021

SECONDARY ACCESSION(S): PXD030188

REPOSITORIES: MassIVE