Abstract / Summary
Abstract Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related death globally, and the lack of predictive preclinical models hampers therapeutic development. Patient-derived organoids (PDOs) recapitulate essential aspects of tumor biology but remain constrained by Matrigel dependence, complex enzymatic processing, loss of the immune microenvironment, and absence of a vascular compartment, which limits nutrient perfusion and drug penetration dynamics. We have established a simplified Matrigel-free method to generate PDOs by mechanically mincing resected tumors into 500–1,000 µm fragments for direct culture. PDOs were established from 17 patients across four major subtypes, with faithful preservation of histological structure, marker expression, somatic mutations, and native stromal and immune components. Drug screening in five PDOs revealed subtype-specific responses: chemo-AK112 outperformed chemo-pembrolizumab in squamous cell carcinoma, whereas chemo-pembrolizumab showed superior efficacy over chemo-AK112 in adenocarcinoma. As a robust preclinical model that preserves the native tumor microenvironment, this Matrigel-free platform enables the dissection of subtype-specific vulnerabilities and facilitates the rational optimization of combination chemo-immunotherapy including in cases that are refractory to standard-of-care regimens.