A Comprehensive Adenoid Cystic Carcinomas-Derived Organoid Platform for Disease Modeling and Drug Screening Captures Interpatient Heterogeneity

Salivary adenoid cystic carcinoma (ACC) is a highly aggressive salivary gland malig-nancy characterized by infiltrative growth patterns that hinder complete resection. Lacking effective chemotherapy, recurrent or metastatic ACC remains clinically incur-able. This research aimed to develop an efficient culture system for ACC organoids, which can preserve tumor heterogeneity and establish a reliable drug screening plat-form. Under our optimized culture conditions, ACC organoids grew rapidly and suc-cessfully recapitulated three characteristic histopathological patterns. Whole-genome sequencing (WGS) further confirmed they mirrored the genomic features of their pa-rental tumors, including significantly mutated genes, non-coding regulatory region mutations, copy number variation, and minor allele frequency. RNA sequencing con-firmed ACC organoids recapitulated the MYB-NFIB fusion gene. At the protein level, these organoids contained multiple cellular components, including epithelial cells, mesenchymal cells, K7+ duct cells, a-SMA+ myoepithelial cells, K5+ basement mem-brane cells, and CD44+ tumor stem cells, with proper spatial distribution patterns. With an 88% success rate, the first ACC organoids platform, incorporating normal sal-ivary gland (SG) organoids as toxicity controls, enabled high-throughput drug testing within two weeks. In conclusion, we develop an efficient culture system for ACC or-ganoids that can preserve tumor heterogeneity and establish a reliable drug screening platform for mechanistic studies and personalized precision therapy research.