Development of Patient-Derived Organoids from NUT Carcinoma: A Robust and Fully Documented Culture System

Abstract

NUT carcinoma is an aggressive, poorly dierentiated malignancy defined by NUTM1 rearrangements. Median survival  remains 6–7 months and preclinical models are essentially nonexistent. Here we report the first patient-derived  organoid platform for this disease. Starting material included resected tumors, pleural eusions, CTguided fine-needle  aspirates, and transbronchial biopsies. Using a defined medium supplemented with Wnt3a, R-spondin-1, Noggin,  FGF10, EGF, bFGF and smallmolecule inhibitors (A83-01, Y-27632, and Losmapimod), we successfully established  long-term cultures from all four specimen types. Organoids retained characteristic nuclear NUT expression (≥80%  positive cells), preserved the original fusion by FISH and NGS, and remained stable for at least 10 passages. Drug testing  against BET inhibitors yielded clear, reproducible dose-response curves. All procedures, quality controls, and banking  steps were performed according to the recently released ISoOR-ISOB international standard. This platform should  facilitate mechanistic studies and drug screening for a tumor that currently lacks tractable models.