High content screening and proteomic analysis identify a kinase inhibitor that rescues pathological phenotypes in a patient-derived model of Parkinson\u2019s disease

Nasia Antoniou,Ioanna Boumpoureka,Ismini Kloukina,Martina Samiotaki,Rebecca Matsas,Maria Xilouri,Regis Grailhe,George Panayotou,Georgia Kouroupi,Era Taoufik,Kanella Prodromidou,Leonidas Stefanis

doi:10.1038/s41531-022-00278-y

Abstract

Combining high throughput screening approaches with induced pluripotent stem cell (iPSC)-based disease modeling represents a promising unbiased strategy to identify therapies for neurodegenerative disorders. Here we applied high content imaging on iPSC-derived neurons from patients with familial Parkinson’s disease bearing the G209A (p.A53T) α-synuclein (αSyn) mutation and launched a screening campaign on a small kinase inhibitor library. We thus identified the multi-kinase inhibitor BX795 that at a single dose effectively restores disease-associated neurodegenerative phenotypes. Proteomics profiling mapped the molecular pathways underlying the protective effects of BX795, comprising a cohort of 118 protein-mediators of the core biological processes of RNA metabolism, protein synthesis, modification and clearance, and stress response, all linked to the mTORC1 signaling hub. In agreement, expression of human p.A53T-αSyn in neuronal cells affected key components of the mTORC1 pathway resulting in aberrant protein synthesis that was restored in the presence of BX795 with concurrent facilitation of autophagy. Taken together, we have identified a promising small molecule with neuroprotective actions as candidate therapeutic for PD and other protein conformational disorders.

Highlights

Parkinson’s disease (PD) is a complex neurodegenerative disorder affecting 2% of the world population over 65 years of age[1]
We identified the multikinase inhibitor BX795 as a compound that exerts a consistent and sustainable beneficial effect on patient-derived p.A53T-neurons
We found that a single treatment with BX795 has long-lasting consequences in supporting neuritic growth, limiting αSyn protein aggregate formation and restoring axonal neuropathology, recorded two weeks after its addition in human p.A53T neurons

Summary

Introduction

Parkinson’s disease (PD) is a complex neurodegenerative disorder affecting 2% of the world population over 65 years of age[1]. The neuropathological hallmark of PD is the presence of intracytoplasmic inclusions in neuronal cell bodies and neurites, respectively termed Lewy bodies and Lewy neurites[4,5]. These are protein aggregates composed mainly of αsynuclein (αSyn), the major protein linked to sporadic PD6. Only symptomatic or palliative treatments are available with none capable to prevent or slow-down disease progression Dopaminereplacement drugs, such as levodopa, which was identified 53 years ago[14], are used to ameliorate motor symptoms and remain the primary and most effective treatment despite the undesired side-effects and deterioration of efficacy with disease progression. Recent advances in patient-derived induced pluripotent stem cell (iPSC)-

Methods

Results

Conclusion