Abstract 3070: Non-small cell lung cancer patient-derived organoids as high-throughput chemotherapy testing platforms
Authors: Manjunath Yariswamy , Suvilesh Kanve Nagaraj , Innokenteva Iuliia , Warren Wesley , Kimchi Eric T. , Staveley-O’Carroll Kevin F. , Li Guangfu , Mitchem Jonathan B. , Kaifi Jussuf T. 
Published: June 2022

Background: Lung cancer is associated with phenotypic, genotypic, and transcriptomic heterogeneity among patients that poses a threat to the development of precision medicine. Despite being considered as potential drug screening platforms, patient-derived xenograft (PDX) models are limited by low rates and prolonged durations of tumor engraftment. To overcome these major limitations, patient-derived organoid (PDO) models have been developed for standard of care drug testing and the outcome of which is driven by differential transcriptome trajectories.

Experimental methods: Single cell suspension from ten surgically resected patient primary non-small cell lung cancer (NSCLC) tissues (adenocarcinoma and squamous cell carcinoma) was mixed with matrigel (growth factor reduced) and cultured in organoid growth medium. Organoids were characterized for structural (Hematoxylin & Eosin staining) and pathological marker (immunohistochemistry for cytokeratin (CK) 5/6, CK7, Napsin A, Thyroid transcription factor-1 (TTF-1) and p40) similarities with primary tumor. Upon passaging, organoids were seeded in triplicates, allowed to acclimate for 24h and subjected to standard of care chemotherapy testing versus vehicle control. PDOs and corresponding primary tumors were subjected to bulk RNA sequencing to understand the transcriptome driven outcome in chemotherapy sensitive versus resistant organoids.

Results: Organoid growth was monitored by light microscopy. PDOs exhibited different growth kinetics indicating the inter-tumor heterogeneity. Time for PDOs to reach 100-200 μm ranged from 6 days to 14 days and exhibited structural features and pathological marker expression similar to their corresponding primary tumors reflecting respective histological subtypes - adenocarcinoma or squamous cell carcinoma. Chemotherapy testing (1 μg/ml Carboplatin plus 0.5 μg/ml Paclitaxel) showed differential responses (day 6 of treatment). Based on the scoring, PDOs were categorized as partially sensitive, very sensitive or resistant (based on organoid growth or regression in presence of chemotherapeutic drugs). Results of RNA sequencing may reveal distinct transcriptomic signatures and molecular pathways reflecting in differential chemoresponses.

Conclusions and future perspectives: PDOs capitulate structural and pathological marker expression of patient primary tumors and serve as versatile, high-throughput drug testing platforms to treat NSCLC patients. Further, identification of key transcriptomic signatures and molecular pathways resulting in chemoresistance will be critical in formulating personal precision medicine to treat NSCLC patients.

Citation Format: Yariswamy Manjunath, Kanve Nagaraj Suvilesh, Iuliia Innokenteva, Wesley Warren, Eric T. Kimchi, Kevin F. Staveley-O’Carroll, Guangfu Li, Jonathan B. Mitchem, Jussuf T. Kaifi. Non-small cell lung cancer patient-derived organoids as high-throughput chemotherapy testing platforms [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3070.