Evaluation of targeted nanoparticles combined with shock waves treatment in colorectal cancer tumoroids

Colorectal cancer (CRC) is the third most common tumor in the world and the second leading cause of cancer-related deaths. Despite improvements in screening and treatment that considerably reduced the occurrence and death rates over the last fifty years, CRC metastases remain a burden, with a survival rate of only 10% among affected patients. Traditional therapies often fail to penetrate effectively the tumor mass and selectively target it. For this purpose, nanoparticle-based treatments have attracted attention as a promising tool for cancer therapy to restrictively target biomarkers over-expressed by tumor, enabling the development of targeted strategies. In this study, zinc-oxide nanoparticles were synthesized and coated with a lipidic shell to enhance their biocompatibility; exploiting the maleimide-thiol chemistry, an optimized protocol was developed to conjugate the YSA peptide, proven for its selective affinity towards the Epha2 receptor, commonly over-expressed in the CRC cells. The protocol showed high binding efficiency for the peptide, offering a powerful tool for future targeted therapeutic approaches. A distinctive strength of this work was the employment of patient-derived tumoroids furnished by IRCCS Istituto di Candiolo biobank, providing a highly faithful preclinical 3D model. Firstly, nanoparticles performance was evaluated in 2D co-cultures comprising healthy colon fibroblasts (CCD18co) and CRC cells to simulate heterogeneous tumor microenvironment. An artificial intelligence for image analysis was employed to recognize and distinguish regions with healthy and cancerous cell populations to assess the impact of the treatment by observing differences in the over-time evolution between the untreated and nanoparticle-treated conditions. Afterwards, tests with the non-targeted and YSA-targeted lipidic nanoconstructs were performed on CCD18co spheroids and CRC tumoroids at different concentrations. The treatment with NPs was combined with sonodynamic therapy consisting of acoustic shock waves to investigate a possible synergistic effect aimed at enhancing the therapeutic response and cytotoxic effect. The combination of nanoparticles and shock waves exhibited promising therapeutic outcomes in the CRC 3D models, suggesting its potential as an innovative strategy for colorectal cancer treatment.