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

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.

Primary Lung Cancer Organoids for Personalized Medicine—Are They Ready for Clinical Use?

“…While the group reported an initial organoid establishment rate of 88%, a long-term culture of pure NSCLC organoids was only successful in 15% of all cases [ 14 ]. Similar success rates were reported by Dijkstra et al [ 16 ] and Yokota et al [ 18 ] who followed the protocol described by Sachs et al but initially abstained from a Nutlin-3a-based selection. In both studies, a remarkable discrepancy between the rates of initial organoid growth and the rates of histologically or genetically confirmed NSCLC organoid establishment was seen.…”

“…In the study by Dijkstra et al, organoid growth was seen in 41% of all cases, but upon histological, immunohistochemical and genomic analysis, the majority of all organoids were normal airway organoids, and pure NSCLC organoids were only found in 17% [ 16 ]. Yokota et al documented organoid growth in 83% of all cases, but only 7% of all organoids were validated as NSCLC [ 18 ]. A number of other medium compositions tested by Yokota et al failed to reach comparable culture success as the airway organoid medium initially reported by Sachs et al [ 18 ]…”

“…Yokota et al documented organoid growth in 83% of all cases, but only 7% of all organoids were validated as NSCLC [ 18 ]. A number of other medium compositions tested by Yokota et al failed to reach comparable culture success as the airway organoid medium initially reported by Sachs et al [ 18 ]…”

“…While most NSCLC organoids were established from tissue of the primary tumor, several groups reported organoids derived from extrapulmonary metastases or pleural effusion [ 11 , 16 , 18 ]. With the lack of competitive growth from normal airway tissue and a potentially more aggressive tumor biology, a higher success in organoid establishment is reported from extrapulmonary NSCLC metastases [ 16 ].…”

“…However, according to The Cancer Genome Atlas (TCGA), up to 15% of adenocarcinomas and 7% of squamous cell carcinomas present with a normal CN profile and are therefore difficult to distinguish from normal airway tissue based on this method [ 16 ]. Other groups used whole-exome sequencing (WES) or targeted sequencing for oncogenic drivers to verify the organoids’ malignant origin [ 11 , 12 , 14 , 15 , 17 , 18 ]. However, due to an inherent genetic drift during organoid passage, genetic alterations or an inconsistency in the present mutations was reported in several cases [ 12 , 18 ].…”