Abstract 205: Patient-derived organoids from surgically treated, localized non-small cell lung cancer as high-throughput drug testing platforms for conventional and repurposed drugs
Authors: Manjunath Yariswamy , Nagaraj Suvilesh Kanve , Nussbaum Yulia I. , Gadelkarim Mohamed , Staveley-O’Carroll Kevin F. , Kimchi Eric T. , Li Guangfu , Warren Wesley , Shyu Chi-Ren , Ciorba Matthew , Mitchem Jonathan B. , Kaifi Jussuf T. 
Published: April 2023

Background: Non-metastatic NSCLC treated with curative surgery has a five-year survival of ~50%, mostly due to development of recurrences. Despite being considered as potential drug screening platforms, patient-derived xenograft (PDX) models are inefficient due to low tumor engraftment rates and complex animal care. Patient-derived organoid (PDO) models overcome these limitations as potential clinically applicable drug testing platforms. Our objective was to develop PDO models from non-metastatic NSCLC patients to study epithelial cell type heterogeneity and drug sensitivities for precision medicine.

Hypothesis: PDO models reliably capitulate the patient primary tumor and serve as versatile platforms for high-throughput screening of standard-of-care and repurposed drugs.

Methods: Single cell suspensions prepared from resected lung tumor tissues from ten NSCLC patients (both adenocarcinoma and squamous cell carcinoma) were mixed with Matrigel (growth factor-reduced) and cultured in organoid growth medium. Organoids and matched primary tumors were compared by histopathology [H&E staining and immunohistochemistry for cytokeratin (CK) 5/6, CK7, Napsin A, Thyroid transcription factor-1 (TTF-1) and p40] and by bulk RNA sequencing. Upon passaging, PDOs were seeded in triplicates, treated with carboplatin/paclitaxel doublet chemotherapy, and drug responses were determined using bright-field 3D imaging (z-stack method). Tumor growth (%) was determined on day 3 and 6 of treatments. RNA sequencing analyses identified a potential drug targets and repurposed drug (aldoketoreductase inhibitor Epalrestat) was tested to overcome chemoresistance in PDOs.

Results: NSCLC PDO growth was established from 11/12 (91.7%) primary tumors with a median time of 11 days (range 4-18 days) to reach volume of 100 μm3. PDOs retained histopathological features and biomarker expression of the matched tumors. As determined by growth differences (p&lt;0.05), 5/9 (55.6%) PDOs were chemosensitive and 4/9 (44.4%) PDOs were chemoresistant against carboplatin/paclitaxel. Whole transcriptome analysis confirmed conservation of the epithelial cell composition in all PDOs in comparison to the matched primary tumors. Potentially druggable genes associated with chemotherapy resistance were identified and the aldoketoreductase inhibitor Epalrestat was repurposed and shown to be effective in overcoming chemoresistance towards carboplatin/paclitaxel.

Conclusions: PDOs can be established from resectable NSCLC patients with high success rates, while preserving histopathological, cellular, and molecular characteristics of the matched primary tumors. Considering the time frames of growth and drug testing, PDOs can serve as clinically applicable drug response testing platforms to traditional and repurposed drugs.

Citation Format: Yariswamy Manjunath, Suvilesh Kanve Nagaraj, Yulia I. Nussbaum, Mohamed Gadelkarim, Kevin F. Staveley-O’Carroll, Eric T. Kimchi, Guangfu Li, Wesley Warren, Chi-Ren Shyu, Matthew Ciorba, Jonathan B. Mitchem, Jussuf T. Kaifi. Patient-derived organoids from surgically treated, localized non-small cell lung cancer as high-throughput drug testing platforms for conventional and repurposed drugs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 205.