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N. Radulovich



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    P1.02 - Biology/Pathology (ID 614)

    • Event: WCLC 2017
    • Type: Poster Session with Presenters Present
    • Track: Biology/Pathology
    • Presentations: 1
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      P1.02-016 - Establishment of Lung Adenocarcinoma Organoid Cultures (ID 9386)

      09:30 - 16:00  |  Author(s): N. Radulovich

      • Abstract

      Background:
      Effective non immune- oncology targeted therapies are available only for less than 25% (non-Asian) and 60% (East Asian) of lung adenocarcinoma (ADC) patients. ADC has one of the largest burdens of genetic abnormalities among all cancers. It is understood that ADC arise from the accumulation of abnormalities which dysregulate key cellular processes to permit a growth and survival. Further improvement in our ability to develop novel therapies requires additional lung cancer models that closer mimic the genetic alterations found in patient tumors. Three-dimensional organoid culture (“mini organs”) of tumor cells recently has generated great interest as such a novel preclinical model.

      Method:
      We experimented to develop novel media formulation to generate organoid models from 10 established ADC cell lines, 7 primary culture ADC cell lines developed from patient-derived xenograft (PDX) models, 8 ADC PDX models, and 20 resected patient ADC tissues. Organoid cultures that could be serially passaged for at least 5 passages were defined as long-term organoid models. Organoids were characterized for their histopathological features and immunomarker expression (p40, TTF-1, p53), growth rate and drug sensitivities.

      Result:
      Long-term organoid cultures were developed from 9/10 (90%) of established ADC cell lines, 3/7 (42%) of PDX-derived cell lines, 2/8 (25%) of ADC PDX models, and 1/20 (5%) of primary patient ADC tissues. Established organoid cultures recapitulated the histological features of ADC. We are currently collecting the data on growth rates and drug sensitivities of selected organoid cultures.

      Conclusion:
      Lung adenocarcinoma organoid cultures can be established for both established cell lines and patient tumor tissues but with variable success rates. Further studies are necessary to understand the discrepancy in the establishment rates from different sources of the tumor cells.

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    P3.03 - Chemotherapy/Targeted Therapy (ID 719)

    • Event: WCLC 2017
    • Type: Poster Session with Presenters Present
    • Track: Chemotherapy/Targeted Therapy
    • Presentations: 1
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      P3.03-008 - Organoid Cultures of Lung Squamous Cell Carcinoma for Drug Screening (ID 8481)

      09:30 - 16:00  |  Author(s): N. Radulovich

      • Abstract

      Background:
      The difficulty of establishing lung squamous cell carcinoma (LUSC) derived cell lines have posed significant challenges for identifying potential therapeutic targets and understanding the complexity of this disease. We have previously developed a LUSC patient-derived xenograft (PDX) platform in which over 50 models have been characterized on the genomic and transcriptomic level. We describe a method to culture and establish LUSC organoids from PDX models and demonstrate their utility for drug testing.

      Method:
      Surgically resected LUSC were implanted into the subcutaneous flank of NOD/SCID mice to establish PDXs. To generate organoids, PDX tissue was dissociated into single cells using Liberase and TypLE and plated in growth factor reduced matrigel dome with media overlay. Organoids were processed for histological and immunohistochemical marker characterization. Organoids and matched PDX were subjected to shallow next generation sequencing for mutation and copy number analysis. Drug screening was performed in 384 well plates and viability was determined by Celltiter Glo 3D assay.

      Result:
      Of the 17 LUSC PDX models attempted, organoid lines from 3 PDX models were propagated beyond 20 passages for over 100 days. The success rate of organoid establishment is 18%, which is higher than establishing LUSC cell lines. Organoids exhibited various doubling rates ranging from 38 to 48 hours. Organoid tumor cells faithfully recapitulated the immuno-phenotypes of the matched PDX, expressing p63 and CK5/6 and were EpCAM positive and H2K negative by flow cytometry analysis. Organoids implanted in NOD/SCID mice formed tumors that reflected the histology of the matched PDX. Shallow sequencing revealed similar copy number status between the organoid and matched PDX. RNA sequencing analysis is pending and will be reported. Organoids were amenable for drug testing and exhibited varying sensitivities to the PI3K inhibitors BKM120 and BYL719 based on each model’s PI3K pathway status.

      Conclusion:
      We describe a method of developing LUSC PDX-derived organoids that can be propagated long term and faithfully recapitulate the histological and molecular characteristics of the original tumor. Additionally, we demonstrate their utility for in vitro drug testing. Organoids may be useful for preclinical modeling and therapeutic evaluation of LUSC.