Integrated clinical and computational data-based repurposing of econazole as a novel autophagic activator in ULK1-related Parkinson disease

Parkinson disease (PD), the second most common neurodegenerative disorder, is pathologically linked to dysregulated Autophagy, a conserved lysosomal degradation pathway. Current conventional PD therapies are often limited by significant side effects, underscoring the demand for alternative treatment strategies. Drug repurposing of FDA-approved compounds represents a promising approach to address this unmet clinical need. Here, by integrating clinical data analysis, we identified an association between Autophagy impairment and specific PD patient subtypes, suggesting that ULK1-dependent Autophagy activation may offer therapeutic benefit. Through systematic screening for Autophagy induction and neuroprotective activity, we identified econazole, a known imidazole Antifungal, as a promising candidate. Econazole exhibited robust therapeutic effects across multiple PD models, including MPTP-induced zebrafish and mouse models, as well as SNCA^A53T mutant mouse models. Notably, its efficacy was dependent on functional Autophagy, as Autophagy inhibition abrogated its beneficial effects. Mechanistically, econazole activated ULK1, enhanced autolysosome formation, and promoted clearance of SNCA aggregates. Mouse brain microarray analysis indicated that econazole-activated ULK1 suppresses MAP3K12/DLK-MAPK8/JNK-MAPK9/JNK2-mediated neuronal Apoptosis. Further phosphoproteomic profiling uncovered a novel ULK1-HSPA8/Hsc70 interaction that promotes LAMP1 and LAMP2 activation and enhances lysosomal function. This ULK1-HSPA8 complex additionally activated the BECN1 (beclin 1) complex to facilitate autophagosome formation. Together, our findings highlight a clinical data-guided drug repurposing approach that identifies econazole as a potent Autophagy activator with therapeutic efficacy in ULK1-linked PD models, opening new avenues for PD treatment.Abbreviations: 3-MA: 3-methyladenine; ACTB: actin beta; ATG: Autophagy related; AUC: area under the curve; BafA1: bafilomycin A1; BECN1: beclin 1; CMA: chaperone-mediated autophagy; DA: dopamine; DOPAC: 3,4-dihydroxyphenylacetic acid; Econ: econazole; GFP: green fluorescent protein; HEK-293T: human embryonic kidney 293T; HSPA8: heat shock protein 8 family A (HSP70) member 8; HVA: homovanillic acid; JUN: Jun proto-oncogene, AP-1 transcription factor subunit; KSEA: kinase-substrate enrichment analysis; LAMP: lysosome associated membrane protein; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MAP2K: mitogen-activated protein kinase kinase; MAP3K12: mitogen-activated protein kinase kinase kinase 12; MAPK: mitogen-activated protein kinase; MPP+: 1-methyl-4-phenylpyridinium; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; PD: Parkinson disease; RB1CC1: RB1 inducible coiled-coil 1; RFP: red fluorescent protein; RMSD: root mean square deviation; SEM: standard error of the mean; SNCA: synuclein alpha; SQSTM1: sequestosome 1; SYP: synaptophysin; TFEB: transcription factor EB; TH: tyrosine hydroxylase; ULK1: unc-51 like Autophagy activating kinase 1; WT: wild-type.