Abstract 3065: Establishment and characterization of a platform of endometrial cancer organoids
Authors: Saldanha Aisha L. , Vo Ha V. , Vasquez Kevin , Ngo Kenneth , Roychoudhury Shrabasti , Feeney Carina , Qi Courtney H. , Narayan Swati , Curtis Jennifer D. , Gokhale Prafulla C. , Chowdhury Dipanjan , Paweletz Cloud P. , Nucci Marisa R. , Matulonis Ursula A. , Ivanova Elena , Liu Joyce F. 
Published: June 2022

Background: Endometrial cancer is the most commonly diagnosed gynecologic cancer in the US; the incidence is rising, and survival rates for this cancer are decreasing. There is a paucity of effective treatment for recurrent endometrial cancer, especially high grade endometrial cancers (HGEC) which include serous, carcinosarcoma, endometrioid, and clear cell histologies. Models that mimic the clinical and molecular characteristics of HGEC are lacking. To support the development of next generation therapeutics for endometrial cancer, we report on the establishment of 3D endometrial patient-derived organoids (PDOs) from HGEC.

Methods: 26 Tumors from 21 different patients with HGEC (Serous, Carcinosarcoma, Clear Cell and High-grade Endometrioid subtypes) who underwent surgical resection (n= 13), biopsy (n = 7), paracentesis (n = 3) or thoracentesis (n = 3) were passaged as 3D organoid cultures in Matrigel in an optimized media. Robust models (defined by average days to passage &lt;14 days) were viably banked. 3 frozen models were also thawed and re-cultured to assess the viability post freezing. PDOs were collected for H&E staining and their histology was compared to the original diagnosis. DNA replication rate and the effect of replication stress on organoid growth were assessed by the DNA Fiber Assay and immunofluorescence (IF). Finally, an established clear cell endometrial cancer organoid model was engrafted in mice to generate a Patient-Derived Xenograft (PDX) model.

Results: Endometrial PDOs were successfully developed from 19 of 26 original samples for an overall success rate of 73.1%. Successful PDOs were developed from multiple histologies, including 8 carcinosarcoma, 6 uterine serous, 2 endometrioid, 2 clear cell and 1 mixed uterine serous and endometrioid. Though biopsy samples had initially fewer viable cells, our overall success rate was similar at 85.7% compared to 84.6% for surgical resections and higher than 66.7% for paracenteses. Samples obtained via thoracentesis did not form PDOs. Endometrial PDOs were histologically validated to match the primary patient tumor. Freeze thawing had no effect on morphology and growth characteristics. DNA fiber assays could be successfully conducted in PDOs, with a reduction in replication rate observed in PDO models treated with ATR or WEE1 inhibitors, with concurrent increase in y-H2AX and decrease in pRPA2 observed by IF. We also successfully generated a validated PDX model from organoids. Studies to determine molecular fidelity between the original patient tumor and established organoids are ongoing.

Conclusions: We describe the successful establishment of 19 endometrial PDO models which retain original tumor morphology and demonstrate sensitivity to drug-induced DNA damage. 3D endometrial organoids can therefore be used for further target discovery and validation as well as biomarker studies to advance targeted therapies for high-grade endometrial cancer.

Citation Format: Aisha L. Saldanha, Ha V. Vo, Kevin Vasquez, Kenneth Ngo, Shrabasti Roychoudhury, Carina Feeney, Courtney H. Qi, Swati Narayan, Jennifer D. Curtis, Prafulla C. Gokhale, Dipanjan Chowdhury, Cloud P. Paweletz, Marisa R. Nucci, Ursula A. Matulonis, Elena Ivanova, Joyce F. Liu. Establishment and characterization of a platform of endometrial cancer organoids [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 3065.