Ultrasound-Responsive Folate-Chlorogenic Acid Nanocarrier Targeting Mixed Lineage Kinase 1 for Enhanced Wilms Tumor Therapy

Abstract

Background:

Wilms tumor (nephroblastoma) is the most frequent malignant tumor of the urinary system in children; however, current treatment choices are restricted due to inadequate drug targeting, systemic toxicity, and multidrug resistance. Chlorogenic acid (CGA), a natural polyphenolic molecule, has promise as an anti-Wilms tumor action but has limited water solubility and bioavailability. To address these constraints, the authors created an ultrasound-responsive folate-CGA nanocarrier (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol-folic acid-CGA [DSPE-PEG-FA/CGA]) that enables targeted and regulated drug administration.

Methods and Results:

The nanocarrier was created utilizing DSPE as the structural backbone, with PEG modification for increased stability and FA conjugation for tumor-specific targeting. Following low-intensity ultrasonic treatment, the nanocarrier increased permeability and induced CGA release via acoustic cavitation effects, greatly enhancing local drug accumulation inside tumor tissues.

Ultrasound Augmentation:

Comprehensive characterization demonstrated good encapsulation efficiency, consistent particle size, and acceptable ultrasonic responsiveness. In vitro experiments indicated increased cellular absorption via folate receptor-mediated endocytosis and ultrasound stimulation, leading to more cytotoxicity against Wilms tumor cells than nonultrasound-treated controls. In vivo, the DSPE-PEG-FA/CGA system demonstrated selective tumor accumulation, dose-dependent tumor growth suppression, and good biosafety in nude mice, with no changes in body weight or organ damage. Mechanistic research revealed mixed-lineage kinase 1 (MLK1) as a direct molecular target of CGA, since it binds to MLK1’s active pocket (specifically residues L303, I228, and A249) and modifies the MAPK and PI3K-Akt signaling pathways to induce apoptosis and decrease proliferation.

Conclusions:

Overall, this work shows that ultrasound-responsive folate-CGA nanotherapy is an effective, safe, and focused method for treating Wilms tumors by combining molecular suppression of MLK1 with ultrasound-enhanced delivery accuracy.