Hui Chen1, Kristin Gotimer1, Cristabelle De Souza2, Clifford G Tepper2, Anthony N Karnezis3, Gary S Leiserowitz1, Jeremy Chien4, Lloyd H Smith5. 1. Department of Obstetrics and Gynecology, University of California Davis Health, Sacramento, CA 95817, United States of America. 2. Department of Biochemistry and Molecular Medicine, University of California Davis Health, Sacramento, CA 95817, United States of America. 3. Department of Pathology and Laboratory Medicine, University of California Davis Health, Sacramento, CA 95817, United States of America. 4. Department of Obstetrics and Gynecology, University of California Davis Health, Sacramento, CA 95817, United States of America; Department of Biochemistry and Molecular Medicine, University of California Davis Health, Sacramento, CA 95817, United States of America. Electronic address: jrchien@ucdavis.edu. 5. Department of Obstetrics and Gynecology, University of California Davis Health, Sacramento, CA 95817, United States of America. Electronic address: lhsmith@ucdavis.edu.
Abstract
OBJECTIVE: Cancer patient-derived organoids (PDOs) grow as three dimensional (3D) structures in the presence of extracellular matrix and have been found to represent the original tumor's genetic complexity. In addition, PDOs can be grown and subjected to drug sensitivity testing in a shorter time course and with lesser expense than patient-derived xenograft models. Many patients with recurrent ovarian cancer develop malignant effusions that become refractory to chemotherapy. Since these same patients often present for palliative aspiration of ascites or pleural effusions, there is a potential opportunity to obtain tumor specimens in the form of multicellular spheroids (MCS) present in malignant effusion fluids. Our objective was to develop a short duration culture of MCS from ovarian cancer malignant effusions in conditions selected to support organoid growth and use them as a platform for empirical drug sensitivity testing. METHODS: In this study, malignant effusion specimens were collected from patients with high-grade serous ovarian carcinoma (HGSOC). MCS were recovered and subjected to culture conditions designed to support organoid growth. In a subset of specimens, RNA-sequencing was performed at two time points during the short-term culture to determine changes in transcriptome in response to culture conditions. Organoid induction was also characterized in these specimens using Ki67 staining and histologic analysis. Drug sensitivity testing was performed on all specimens. RESULTS: Our model describes organoids formed within days of primary culture, which can recapitulate the histological features of malignant ascites fluid and can be expanded for at least 6 days. RNA-seq analysis of four patient specimens showed that within 6 days of culture, there was significant up-regulation of genes related to cellular proliferation, epithelial-mesenchymal transition, and KRAS signaling pathways. Drug sensitivity testing identified several agents with therapeutic potential. CONCLUSIONS: Short duration organoid culture of MCS from HGSOC malignant effusions can be used as a platform for empiric drug sensitivity testing. These ex vivo models may be helpful in screening new or existing therapeutic agents prior to individualized treatment options.
OBJECTIVE:Cancerpatient-derived organoids (PDOs) grow as three dimensional (3D) structures in the presence of extracellular matrix and have been found to represent the original tumor's genetic complexity. In addition, PDOs can be grown and subjected to drug sensitivity testing in a shorter time course and with lesser expense than patient-derived xenograft models. Many patients with recurrent ovarian cancer develop malignant effusions that become refractory to chemotherapy. Since these same patients often present for palliative aspiration of ascites or pleural effusions, there is a potential opportunity to obtain tumor specimens in the form of multicellular spheroids (MCS) present in malignant effusion fluids. Our objective was to develop a short duration culture of MCS from ovarian cancer malignant effusions in conditions selected to support organoid growth and use them as a platform for empirical drug sensitivity testing. METHODS: In this study, malignant effusion specimens were collected from patients with high-grade serous ovarian carcinoma (HGSOC). MCS were recovered and subjected to culture conditions designed to support organoid growth. In a subset of specimens, RNA-sequencing was performed at two time points during the short-term culture to determine changes in transcriptome in response to culture conditions. Organoid induction was also characterized in these specimens using Ki67 staining and histologic analysis. Drug sensitivity testing was performed on all specimens. RESULTS: Our model describes organoids formed within days of primary culture, which can recapitulate the histological features of malignant ascites fluid and can be expanded for at least 6 days. RNA-seq analysis of four patient specimens showed that within 6 days of culture, there was significant up-regulation of genes related to cellular proliferation, epithelial-mesenchymal transition, and KRAS signaling pathways. Drug sensitivity testing identified several agents with therapeutic potential. CONCLUSIONS: Short duration organoid culture of MCS from HGSOC malignant effusions can be used as a platform for empiric drug sensitivity testing. These ex vivo models may be helpful in screening new or existing therapeutic agents prior to individualized treatment options.
Authors: Sarah J Hill; Brennan Decker; Emma A Roberts; Neil S Horowitz; Michael G Muto; Michael J Worley; Colleen M Feltmate; Marisa R Nucci; Elizabeth M Swisher; Huy Nguyen; Chunyu Yang; Ryuji Morizane; Bose S Kochupurakkal; Khanh T Do; Panagiotis A Konstantinopoulos; Joyce F Liu; Joseph V Bonventre; Ursula A Matulonis; Geoffrey I Shapiro; Ross S Berkowitz; Christopher P Crum; Alan D D'Andrea Journal: Cancer Discov Date: 2018-09-13 Impact factor: 39.397
Authors: Hiu Wing Cheung; Glenn S Cowley; Barbara A Weir; Jesse S Boehm; Scott Rusin; Justine A Scott; Alexandra East; Levi D Ali; Patrick H Lizotte; Terence C Wong; Guozhi Jiang; Jessica Hsiao; Craig H Mermel; Gad Getz; Jordi Barretina; Shuba Gopal; Pablo Tamayo; Joshua Gould; Aviad Tsherniak; Nicolas Stransky; Biao Luo; Yin Ren; Ronny Drapkin; Sangeeta N Bhatia; Jill P Mesirov; Levi A Garraway; Matthew Meyerson; Eric S Lander; David E Root; William C Hahn Journal: Proc Natl Acad Sci U S A Date: 2011-07-11 Impact factor: 11.205
Authors: Vladimir J N Bykov; Nicole Zache; Helene Stridh; Jacob Westman; Jan Bergman; Galina Selivanova; Klas G Wiman Journal: Oncogene Date: 2005-05-12 Impact factor: 9.867
Authors: Paulina Cybulska; Jocelyn M Stewart; Azin Sayad; Carl Virtanen; Patricia A Shaw; Blaise Clarke; Natalie Stickle; Marcus Q Bernardini; Benjamin G Neel Journal: Am J Pathol Date: 2018-02-16 Impact factor: 4.307
Authors: Justin W Gorski; Zhuwei Zhang; J Robert McCorkle; Jodi M DeJohn; Chi Wang; Rachel W Miller; Holly H Gallion; Charles S Dietrich; Frederick R Ueland; Jill M Kolesar Journal: Biomedicines Date: 2021-08-16
Authors: Cristabelle De Souza; Jill Madden; Devin C Koestler; Dennis Minn; Dennis J Montoya; Kay Minn; Alan G Raetz; Zheng Zhu; Wen-Wu Xiao; Neeki Tahmassebi; Harikumara Reddy; Nina Nelson; Anthony N Karnezis; Jeremy Chien Journal: J Natl Cancer Inst Date: 2021-09-04 Impact factor: 13.506