PO-271 Using human lung organoids to study pulmonary neuroendocrine cells

Abstract

Introduction Introduction: Lung neuroendocrine tumours (LNETs) arise from a rare lung cell population, called pulmonary neuroendocrine cells (PNECs), and account for 20%–25% of all lung cancers. Despite the involvement of PNECs in LNETs and a number of other respiratory diseases, relatively little is known about their normal function or contribution to cancer development and progression. To date, the study of PNECs has been limited by difficulties in isolating and culturing this cell population. Material and methods Building on methods developed in our lab for the isolation and long-term expansion of healthy and diseased human airway cells, we have generated lung organoids derived from embryonic lung tissue. We have identified cultures conditions that allow for the enrichment of PNECs in embryonic lung organoids. In parallel, we have begun to generate a biobank of both low- and high-grade LNET organoids. Results PNEC-enriched embryonic lung organoids can be expanded indefinitely over multiple passages. Preliminary analysis by qPCR and whole mount immunofluorescence shows that PNEC-enriched organoids express a number of PNEC lineage markers including ASCL1, NEUROD1, and UCHL1. PNEC-enriched lung organoids also express PNEC-associated bioactive compounds such as SST and CGRP. Conclusions We have generated human embryonic lung organoids that contain significant numbers of PNECs. Applying the same principles, we have established organoid cultures of human LNETs. PNEC-enriched organoids and LNET organoids are currently being used for further molecular and genetic analyses.