3D Bioprinted Vascularized Lung Cancer Organoid Models Recapitulating Tumor Microenvironment with Underlying Diseases for Implementing Reliable Drug Efficacy Tests

Abstract

Despite the promising advancements in in vitro cancer models, the development of comprehensive in vitro cancer models that faithfully replicate the intricate tumor microenvironment (TME), encompassing diverse cellular components and genetic properties, remains insufficient. In this study, I present an advanced vascularized lung cancer model that integrates patient-derived lung cancer organoids (LCOs), lung fibroblasts, and perfused vessels utilizing 3D bioprinting technology. My primary objective is to establish an in vitro lung cancer model that similarly mimics the patient's TME and enables precise drug evaluation by constructing a vascularized LCO model incorporating the underlying disease. To enhance their physiological resemblance to native lung tissue and provide physical and biochemical cues for cells within the TME, I generated lung tissue-derived decellularized extracellular matrix (LudECM) hydrogels. Two distinct bioinks were developed, including LCOs and idiopathic pulmonary fibrosis-derived lung fibroblasts (iLFs). Remarkably, the LudECM hydrogels derived from lung tissue were more favorable than Matrigel in terms of signal stimulation and support for the necessary biological activities essential for cell growth and functionality within the TME. The LCOs cultured on LudECM hydrogels successfully maintained patient-specific genetic mutation characteristics. In addition, iLFs were effective to produce fibrotic niches similar to human fibrosis. To recapitulate lung cancer with fibrosis, LCOs and iLFs were co-cultured within LudECM, and the drug responsiveness of LCOs to targeted anticancer drugs was evaluated. The LCOs co-cultured with iLFs resulted in variable resistance to sensitizing drugs, such as poziotinib, as opposed to resistant drugs. Indeed, it was confirmed that gene expression levels related to drug resistance increased in LCOs with fibrosis. Finally, the LCOs exhibited enhanced proliferation in models integrating perfused vessels with fibrosis, and a higher number of proliferating cancer cells existed within the LCOs accompanied by fibrosis when the drug was administered via the vessel. As a result, the in vitro vascularized LCOs model, which faithfully recapitulates lung cancer environment with pulmonary fibrosis and allows for the assessment of drug response, holds tremendous potential in guiding treatment decisions for lung cancer patients with underlying diseases. Furthermore, this approach has the prospect to facilitate the development of targeted therapies and the discovery of biomarkers for lung cancer patients with underlying diseases.