Report

Teaser, summary, work performed and final results

Periodic Reporting for period 2 - LYSOSOMICS (Functional Genomics of the Lysosome)

Teaser

Lysosomal Storage Disorders (LSDs) are a group of approximately 50 inherited diseases with progressive, multisystemic symptoms, mostly affecting children. Although great advances have been done in LSDs research, current therapeutic options are inefficient or not available. The...

Summary

Lysosomal Storage Disorders (LSDs) are a group of approximately 50 inherited diseases with progressive, multisystemic symptoms, mostly affecting children. Although great advances have been done in LSDs research, current therapeutic options are inefficient or not available. The overall objective of the study is to generate genetically modified cell lines to study the lysosomal function, resembling the pathological cellular environment of many diseases and to use them as a platform for the identification of new therapeutic approaches.

Work performed

We generated novel cellular models for all known LSDs by engineering cell lines using the CRISPR-Cas9 technology. In this year, we obtained a set of 18 different cell lines. A preliminary characterization revealed that most of the cell lines generated using this approach, reproduce the disease phenotypes. These cell lines show lysosomal dysfunction, that is a typical hallmark of LSDs. This dysfunction was rescued by reintroducing the wild type gene. We also performed multiparametric experiment for the simultaneous analysis of lysosomal morphology and function, demonstrating that these cell lines are suitable for drugs screening experiments. The generation of a new LSDs Biobank (LSD-CRIB) showed that these new cells lines are a powerful tool to perform comparative and mechanistic studies of lysosomal physiology, dynamics, positioning. Moreover, we demonstrated that these cell lines can be used for drug screening approaches.

Final results

We were the first group to create a biobank of such a high number of LSDs cellular models that can be broadly used as a platform for the implementation of new therapeutic approaches. We aim to use this powerful tool to deeply characterize lysosomal genes function, identify new signaling pathways involved in lysosomal regulation.