Abstract 469: Patient-derived sarcoma models: First results from the SARQMA study
Authors: Gaebler Manuela , Silvestri Alessandra , Reichardt Peter , Wardelmann Eva , Gambara Guido , Haybaeck Johannes , Stroebel Philipp , Niethard Maya , Erdmann Gerrit , Regenbrecht Christian R. 
Published: August 2019

Objective: The development of new targeted therapeutics has allowed an essential improvement in carcinoma treatment. For sarcomas, however, the main approach is still the combination of surgery, chemotherapy and radiation, which is due to their high heterogeneity, with more than 70 histopathological subtypes, and the limited knowledge of the molecular drivers of tumor development and progression. Here we show that patient-derived 3D (PD3D) cell culture models allow for an in vitro system to systematically test compounds and combos in a semi-automated way, generating a pre-clinical dataset that in combination with clinical data, genomic and proteomics profiles may help to better understand the biology and ultimately identify more potent treatment regimens.

Methods: Fresh surgical specimen underwent several steps of dissociation. Cell aggregates were then seeded into 24w plates in matrix-like scaffolds and grown until &gt;100μm. Organoids were then harvested, transferred to 384w plates and treated with a set of compounds that resemble standard-of-care treatments and novel compounds. Finally, viability was calculated. In parallel, protein extracts were used for DigiWest, a multiplexed protein profiling assay allowing to interrogate up to 800 (phospho)-proteins.

Results: Of 49 cases with a biopsy, 23 had too little material available. Of these 23 samples, 5 were reported as sarcoma tissue afterwards, while 5 of the taken 26 samples were not sarcomas according to the final histopathology. Of all sarcomas, 90% (19/21) were taken from tumors localized at the extremities, the rest was located at the trunk. 52% (11/21) were growing in short term cell culture at least to passage 1 (p1), 28% (6/21) were growing for long term analyses. Two of those six were myxoid liposarcomas, two were undifferentiated pleomorphic sarcomas or classified as not otherwise specified (UPS/NOS), one was a myxoid liposarcoma and one a biphasic synovial sarcoma. Here we show that it is feasible to generate organoids from sarcoma tissues for extensive characterization in order to better understand their biology and mechanisms of treatment. High-throughput drug screening allows for an profiling of pharmacokinetic properties of individual sarcomas. Using the same material for additional (phospho)-proteomics provides multiple layers of understanding.

Conclusion: At present, the structure of clinical trials is not amenable to N of 1 studies, so applying the information garnered from this platform, particularly combination therapy drug screens, remains a significant hurdle. The major limitation to the establishment of organoid cultures was insufficient amounts of fresh tissue with viable tumor cells. Increasing the tumor tissue available for organoid production would lead to a greater success rate. Nevertheless, by their recapitulation of the donor tissue architecture, they provide an interesting and important tool to study the huge variety of soft-tissue tumors.

Citation Format: Manuela Gaebler, Alessandra Silvestri, Peter Reichardt, Eva Wardelmann, Guido Gambara, Johannes Haybaeck, Philipp Stroebel, Maya Niethard, Gerrit Erdmann, Christian R. Regenbrecht. Patient-derived sarcoma models: First results from the SARQMA study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 469.