Clinical application of a lung cancer organoid (tumoroid) culture system

Etsuko Yokota,Miki Iwai,Yoshio Naomoto,Yutaka Maeda,Yasumasa Monobe,Minzhe Guo,Nagio Takigawa,Minoru Haisa,Takuya Fukazawa,Tomoki Yamatsuji,Masakazu Yoshida,Takuro Yukawa

doi:10.1038/s41698-021-00166-3

Abstract

Despite high expectations for lung tumoroids, they have not been applied in the clinic due to the difficulty of their long-term culture. Here, however, using AO (airway organoid) media developed by the Clevers laboratory, we succeeded in generating 3 lung tumoroid lines for long-term culture (>13 months) from 41 lung cancer cases (primary or metastatic). Use of nutlin-3a was key to selecting lung tumoroids that harbor mutant p53 in order to eliminate normal lung epithelial organoids. Next-generation sequencing (NGS) analysis indicated that each lung tumoroid carried BRAFG469A, TPM3-ROS1 or EGFRL858R/RB1E737*, respectively. Targeted therapies using small molecule drugs (trametinib/erlotinib for BRAFG469A, crizotinib/entrectinib for TPM3-ROS1 and ABT-263/YM-155 for EGFRL858R/RB1E737*) significantly suppressed the growth of each lung tumoroid line. AO media was superior to 3 different media developed by other laboratories. Our experience indicates that long-term lung tumoroid culture is feasible, allowing us to identify NGS-based therapeutic targets and determine the responsiveness to corresponding small molecule drugs.

Highlights

The treatment of lung cancer has been dramatically improved in the past 17 years with the discovery of driver oncogenes and the development of molecularly targeted drugs that bind them, including osimertinib for mutant EGFR or crizotinib for ROS1 fusions[1,2]
The success rate for generating lung tumoroids was only 7% (3/41). These results were consistent with the results reported by Swanton, Voest and colleagues (8% and 17%, respectively)[16,17]
The major reason for this low success rate was because the current tumoroid culture systems allow lung organoids from normal lung epithelial cells to grow faster than lung tumoroids

Summary

Introduction

The treatment of lung cancer has been dramatically improved in the past 17 years with the discovery of driver oncogenes and the development of molecularly targeted drugs that bind them, including osimertinib for mutant EGFR or crizotinib for ROS1 fusions[1,2]. Patients with BRAFV600E, which is seen in melanoma, colorectal cancer and non-small cell lung cancer (NSCLC), are currently treated using vemurafenib in the clinic; there are no molecularly targeted drugs to treat patients with BRAFG469A, which is occasionally seen in NSCLC3. Such cancers treated with small molecule drugs, including osimertinib, crizotinib and vemurafenib, always recur by acquiring different drugresistance mechanisms. The generation of an avatar cancer cell line derived from each patient’s cancer will meet the need for personalized medicine to identify small molecule drugs for each patient so as to eradicate lung cancer cells that harbor previously untargeted driver oncogenes and/or drug-resistance pathways. In vivo patient-derived xenografts (PDXs) using mice have been applied for the same purpose

Methods

Results

Conclusion