Abstract 3079: Development of an automated platform for screening patient-derived organoid models

• Published in: Cancer research (Chicago, Ill.) Ill.), 2022-06, Vol.82 (12_Supplement), p.3079-3079
• Main Authors: Paul, Siddhartha, Hose, Curtis, Jones, Eric, Harris, Erik, Connelly, John, Campbell, Petreena, Ortiz, Mariaestela, Dexheimer, Thomas S., Silvers, Thomas, Brady, Penny Sellers, Grams, Julie, Rohrer, Tiffany Nikirk, Martin, Karen, Ramsey, Patricia, Bowles, Lori, Rapisarda, Annamaria, Parchment, Ralph E., Teicher, Beverly A., Doroshow, James H., Coussens, Nathan P.
• Format: Article
• Language: English
• ISSN: 1538-7445; 1538-7445
• DOI: 10.1158/1538-7445.AM2022-3079

One of the major challenges in preclinical cancer therapeutic development is establishing physiologically relevant in vitro assays that correlate with the in vivo responses of patient tumors to anticancer agents. By incorporating tumor cell heterogeneity and three-dimensional morphological features, patient-derived organoids provide an improved in vivo relevancy compared to established tumor cell lines grown as monolayers. However, organoids grow while embedded in an extracellular matrix material with complex media formulations, which poses challenges for culture scale-up and automated drug screening methods. Organoid models derived from a variety of human solid tumor types are distributed by the National Cancer Institute’s Patient-Derived Models Repository program (https://pdmr.cancer.gov). A panel of human patient-derived colon adenocarcinoma organoid models was assembled to evaluate an automated high-throughput screening (HTS) platform. The organoid panel members were characterized for their reproducible growth and expansion capacity in culture, recovery from cryopreservation, and amenability to operations associated with HTS. Short tandem repeat profiling was performed regularly throughout the process to authenticate each sample. Among the organoid models, variations were observed in morphology (assessed by brightfield imaging) and growth rate (measured by population doublings). Most models expanded well in culture for greater than sixty days and all models demonstrated a sufficient recovery from cryopreservation. The aims in adapting organoid cultures to a HTS platform included minimizing the operational complexity, maximizing the process throughput, and maintaining high organoid viability. Assay conditions for all panel members were selected in conjunction with automated methods, instrumentation, and endpoint measurements. Details such as the optimal sample preparation steps, media formulation, and inoculation density varied among the organoid models. However, other aspects such as liquid handling procedures for organoid inoculation and drug delivery, microwell plate type, assay duration, and endpoint measurements were selected for their suitability to all organoid models tested. Using a custom-designed automated screening system, the refined methods were validated by screening the panel of patient-derived colon adenocarcinoma organoids against a library of oncology drugs approved by the United States Food and Drug Administration (https://dtp.cancer.