Abstract 3079: Development of an automated platform for screening patient-derived organoid models
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. 
Published: June 2022

Abstract
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.gov/organization/dscb/obtaining/default.htm). Assay performance metrics and pharmacological data demonstrate the robust performance of this organoid screening platform. Future efforts will establish additional patient-derived organoid panels for expanded HTS using this platform. This project was funded in part with federal funds from the NCI, NIH, under contract no. HHSN261201500003I.
Citation Format: Siddhartha Paul, Curtis Hose, Eric Jones, Erik Harris, John Connelly, Petreena Campbell, Mariaestela Ortiz, Thomas S. Dexheimer, Thomas Silvers, Penny Sellers Brady, Julie Grams, Tiffany Nikirk Rohrer, Karen Martin, Patricia Ramsey, Lori Bowles, Annamaria Rapisarda, Ralph E. Parchment, Beverly A. Teicher, James H. Doroshow, Nathan P. Coussens. Development of an automated platform for screening patient-derived organoid models [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 3079.