Steven D Forsythe1,2,3, Hemamylammal Sivakumar1,4, Richard A Erali1,3,5,6, Nadeem Wajih1,3, Wencheng Li7, Perry Shen5,6, Edward A Levine5,6, Katherine E Miller8,9, Aleksander Skardal10,11,12,13, Konstantinos I Votanopoulos14,15,16,17,18. 1. Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC, USA. 2. Department of Cancer Biology, Wake Forest School of Medicine, Winston-Salem, NC, USA. 3. Wake Forest Organoid Research Center (WFORCE), Winston-Salem, NC, USA. 4. Department of Biomedical Engineering, The Ohio State University, Columbus, OH, USA. 5. Department of Surgery, Division of Surgical Oncology, Wake Forest Baptist Health, Wake Forest University, Winston-Salem, NC, USA. 6. Comprehensive Cancer Center, Wake Forest School of Medicine, Winston-Salem, NC, USA. 7. Department of Pathology, Wake Forest School of Medicine, Winston-Salem, NC, USA. 8. The Steve and Cindy Rasmussen Institute for Genomic Medicine, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, OH, USA. 9. Department of Pediatrics, The Ohio State University College of Medicine, Columbus, OH, USA. 10. Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC, USA. skardal.1@osu.edu. 11. Department of Cancer Biology, Wake Forest School of Medicine, Winston-Salem, NC, USA. skardal.1@osu.edu. 12. Department of Biomedical Engineering, The Ohio State University, Columbus, OH, USA. skardal.1@osu.edu. 13. The Ohio State University and Arthur G. James Comprehensive Cancer Center, Columbus, OH, USA. skardal.1@osu.edu. 14. Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC, USA. kvotanop@wakehealth.edu. 15. Department of Cancer Biology, Wake Forest School of Medicine, Winston-Salem, NC, USA. kvotanop@wakehealth.edu. 16. Wake Forest Organoid Research Center (WFORCE), Winston-Salem, NC, USA. kvotanop@wakehealth.edu. 17. Department of Surgery, Division of Surgical Oncology, Wake Forest Baptist Health, Wake Forest University, Winston-Salem, NC, USA. kvotanop@wakehealth.edu. 18. Comprehensive Cancer Center, Wake Forest School of Medicine, Winston-Salem, NC, USA. kvotanop@wakehealth.edu.
Abstract
INTRODUCTION: Sarcoma clinical outcomes have been stagnant for decades due to heterogeneity of primaries, lack of comprehensive preclinical models, and rarity of disease. We hypothesized that engineering hydrogel-based sarcoma organoids directly from the patient without xenogeneic extracellular matrices (ECMs) or growth factors is routinely feasible and allows rare tumors to remain viable as avatars for personalized research. METHODS: Surgically resected sarcomas (angiosarcomas, leiomyosarcoma, gastrointestinal stromal tumor, liposarcoma, myxofibrosarcoma, dermatofibrosarcoma protuberans [DFSP], and pleiomorphic abdominal sarcoma) were dissociated and incorporated into a hyaluronic acid and collagen-based ECM hydrogel and screened for chemotherapy efficacy. A subset of organoids was enriched with a patient-matched immune system for screening of immunotherapy efficacy (iPTOs). Response to treatment was assessed using LIVE/DEAD staining and metabolic assays. RESULTS: Sixteen sarcomas were biofabricated into three-dimensional (3D) patient-specific sarcoma organoids with a 100% success rate. Average time from organoid development to initiation of drug testing was 7 days. Enrichment of organoids with immune system components derived from either peripheral blood mononuclear cells or lymph node cells was performed in 10/16 (62.5%) patients; 4/12 (33%) organoids did not respond to chemotherapy, while response to immunotherapy was observed in 2/10 (20%) iPTOs. CONCLUSIONS: A large subset of sarcoma organoids does not exhibit response to chemotherapy or immunotherapy, as currently seen in clinical practice. Routine development of sarcoma hydrogel-based organoids directly from the operating room is a feasible platform, allowing for such rare tumors to remain viable for personalized translational research.
INTRODUCTION: Sarcoma clinical outcomes have been stagnant for decades due to heterogeneity of primaries, lack of comprehensive preclinical models, and rarity of disease. We hypothesized that engineering hydrogel-based sarcoma organoids directly from the patient without xenogeneic extracellular matrices (ECMs) or growth factors is routinely feasible and allows rare tumors to remain viable as avatars for personalized research. METHODS: Surgically resected sarcomas (angiosarcomas, leiomyosarcoma, gastrointestinal stromal tumor, liposarcoma, myxofibrosarcoma, dermatofibrosarcoma protuberans [DFSP], and pleiomorphic abdominal sarcoma) were dissociated and incorporated into a hyaluronic acid and collagen-based ECM hydrogel and screened for chemotherapy efficacy. A subset of organoids was enriched with a patient-matched immune system for screening of immunotherapy efficacy (iPTOs). Response to treatment was assessed using LIVE/DEAD staining and metabolic assays. RESULTS: Sixteen sarcomas were biofabricated into three-dimensional (3D) patient-specific sarcoma organoids with a 100% success rate. Average time from organoid development to initiation of drug testing was 7 days. Enrichment of organoids with immune system components derived from either peripheral blood mononuclear cells or lymph node cells was performed in 10/16 (62.5%) patients; 4/12 (33%) organoids did not respond to chemotherapy, while response to immunotherapy was observed in 2/10 (20%) iPTOs. CONCLUSIONS: A large subset of sarcoma organoids does not exhibit response to chemotherapy or immunotherapy, as currently seen in clinical practice. Routine development of sarcoma hydrogel-based organoids directly from the operating room is a feasible platform, allowing for such rare tumors to remain viable for personalized translational research.
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