TY - JOUR
T1 - Organoids with cancer stem cell-like properties secrete exosomes and HSP90 in a 3D nanoenvironment
AU - Eguchi, Takanori
AU - Sogawa, Chiharu
AU - Okusha, Yuka
AU - Uchibe, Kenta
AU - Iinuma, Ryosuke
AU - Ono, Kisho
AU - Nakano, Keisuke
AU - Murakami, Jun
AU - Itoh, Manabu
AU - Arai, Kazuya
AU - Fujiwara, Toshifumi
AU - Namba, Yuri
AU - Murata, Yoshiki
AU - Ohyama, Kazumi
AU - Shimomura, Manami
AU - Okamura, Hirohiko
AU - Takigawa, Masaharu
AU - Nakatsura, Tetsuya
AU - Kozaki, Ken ichi
AU - Okamoto, Kuniaki
AU - Calderwood, Stuart K.
N1 - Funding Information:
JSR Life Sciences Corporation provided support in the form of salaries for authors RI, MI, and KA, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. This work was supported by JSPS KAKENHI, grant numbers JP17K11642 (to TE), JP16K11722 (to JM TE), JP17K11669 (to CS TE), and JP17K11669 (to KO CS TE), a JSR joint research program (to TE), and a Ryobi Teien Memorial Foundation Grant (to TE). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Publisher Copyright:
Copyright: © 2018 Eguchi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2018/2
Y1 - 2018/2
N2 - Ability to form cellular aggregations such as tumorspheres and spheroids have been used as a morphological marker of malignant cancer cells and in particular cancer stem cells (CSC). However, the common definition of the types of cellular aggregation formed by cancer cells has not been available. We examined morphologies of 67 cell lines cultured on three dimensional morphology enhancing NanoCulture Plates (NCP) and classified the types of cellular aggregates that form. Among the 67 cell lines, 49 cell lines formed spheres or spheroids, 8 cell lines formed grape-like aggregation (GLA), 8 cell lines formed other types of aggregation, and 3 cell lines formed monolayer sheets. Seven GLA-forming cell lines were derived from adenocarcinoma among the 8 lines. A neuroendocrine adenocarcinoma cell line PC-3 formed asymmetric GLA with ductal structures on the NCPs and rapidly growing asymmetric tumors that metastasized to lymph nodes in immunocompromised mice. In contrast, another adenocarcinoma cell line DU-145 formed spheroids in vitro and spheroid-like tumors in vivo that did not metastasize to lymph nodes until day 50 after transplantation. Culture in the 3D nanoenvironment and in a defined stem cell medium enabled the neuroendocrine adenocarcinoma cells to form slowly growing large organoids that expressed multiple stem cell markers, neuroendocrine markers, intercellular adhesion molecules, and oncogenes in vitro. In contrast, the more commonly used 2D serum-contained environment reduced intercellular adhesion and induced mesenchymal transition and promoted rapid growth of the cells. In addition, the 3D stemness nanoenvironment promoted secretion of HSP90 and EpCAM-exosomes, a marker of CSC phenotype, from the neuroendocrine organoids. These findings indicate that the NCP-based 3D environment enables cells to form stem cell tumoroids with multipotency and model more accurately the in vivo tumor status at the levels of morphology and gene expression.
AB - Ability to form cellular aggregations such as tumorspheres and spheroids have been used as a morphological marker of malignant cancer cells and in particular cancer stem cells (CSC). However, the common definition of the types of cellular aggregation formed by cancer cells has not been available. We examined morphologies of 67 cell lines cultured on three dimensional morphology enhancing NanoCulture Plates (NCP) and classified the types of cellular aggregates that form. Among the 67 cell lines, 49 cell lines formed spheres or spheroids, 8 cell lines formed grape-like aggregation (GLA), 8 cell lines formed other types of aggregation, and 3 cell lines formed monolayer sheets. Seven GLA-forming cell lines were derived from adenocarcinoma among the 8 lines. A neuroendocrine adenocarcinoma cell line PC-3 formed asymmetric GLA with ductal structures on the NCPs and rapidly growing asymmetric tumors that metastasized to lymph nodes in immunocompromised mice. In contrast, another adenocarcinoma cell line DU-145 formed spheroids in vitro and spheroid-like tumors in vivo that did not metastasize to lymph nodes until day 50 after transplantation. Culture in the 3D nanoenvironment and in a defined stem cell medium enabled the neuroendocrine adenocarcinoma cells to form slowly growing large organoids that expressed multiple stem cell markers, neuroendocrine markers, intercellular adhesion molecules, and oncogenes in vitro. In contrast, the more commonly used 2D serum-contained environment reduced intercellular adhesion and induced mesenchymal transition and promoted rapid growth of the cells. In addition, the 3D stemness nanoenvironment promoted secretion of HSP90 and EpCAM-exosomes, a marker of CSC phenotype, from the neuroendocrine organoids. These findings indicate that the NCP-based 3D environment enables cells to form stem cell tumoroids with multipotency and model more accurately the in vivo tumor status at the levels of morphology and gene expression.
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U2 - 10.1371/journal.pone.0191109
DO - 10.1371/journal.pone.0191109
M3 - Article
C2 - 29415026
AN - SCOPUS:85041493604
VL - 13
JO - PLoS One
JF - PLoS One
SN - 1932-6203
IS - 2
M1 - e0191109
ER -