# Quantitative Proteomics Unveils the Synergistic Effects of Combination Drugs on Cytoskeleton Composition and Autophagy-Mediated Cell Death in Neuroblastoma

## TL;DR

This study shows that combining two FDA-approved drugs improves neuroblastoma treatment by altering cell structure and increasing cell death.

## Contribution

The study identifies a novel combination therapy for neuroblastoma and reveals its molecular mechanisms through proteomics.

## Key findings

- Combining pyrvinium pamoate and sirolimus reduced cytoskeleton formation and cell migration.
- The drug combination increased autophagy and caused cell cycle arrest in neuroblastoma cells.
- Quantitative proteomics identified 3416 proteins linked to the therapeutic effects of the combination.

## Abstract

Neuroblastoma, a prevalent and aggressive childhood cancer,
lacks
effective treatments. Recent research highlights the repurposing of
existing drugs as a strategy for breakthroughs in combating this disease.
We systematically analyzed small-molecule perturbation gene expression
data from the Library of Integrated Network-Based Cellular Signatures
(LINCS), identifying pyrvinium pamoate and sirolimus, two FDA-approved
drugs, as potential candidates for neuroblastoma combination therapy.
Colony formation assays and organoid culture confirmed that the therapeutic
effect of combining these two drugs exceeded that of either drug alone.
The mRNA expression levels of several genes predicted by LINCS also
decreased. To comprehensively understand the mechanism behind superior
efficacy of the combination therapy compared to monotherapy, we performed
quantitative proteomics with tandem mass tag labeling and identified
3416 proteins from 20,623 peptides. Gene set enrichment analysis and
Database for Annotation, Visualization, and Integrated Discovery revealed
that combination therapy significantly decreased cytoskeleton formation
compared with monotherapy, reflecting dramatic reduction in cell migration.
Additionally, the research indicated that cell cycle arrest occurred
under combination therapy. Furthermore, we confirmed that the extent
of autophagy significantly increased after the combination treatment.
In summary, this study elucidates the mechanisms and therapeutic potential
of combining sirolimus and pyrvinium pamoate for treating neuroblastoma,
offering new advancements for this challenging disease.