Novel bone microenvironment model of castration‑resistant prostate cancer with chitosan fiber matrix and osteoblasts

Samoto,Masahiro; Matsuyama,Hideyasu; Matsumoto,Hiroaki; Hirata,Hiroshi; Ueno,Koji; Ozawa,Sho; Mori,Junichi; Inoue,Ryo; Yano,Seiji; Yamamoto,Yoshiaki; Haginaka,Jun; Horiyama,Shizuyo; Tamada,Koji

doi:10.3892/ol.2021.12950

Novel bone microenvironment model of castration‑resistant prostate cancer with chitosan fiber matrix and osteoblasts

Authors:
- Masahiro Samoto
- Hideyasu Matsuyama
- Hiroaki Matsumoto
- Hiroshi Hirata
- Koji Ueno
- Sho Ozawa
- Junichi Mori
- Ryo Inoue
- Seiji Yano
- Yoshiaki Yamamoto
- Jun Haginaka
- Shizuyo Horiyama
- Koji Tamada
View Affiliations

Affiliations: Department of Urology, Graduate School of Medicine, Yamaguchi University, Ube, Yamaguchi 755‑8505, Japan, Center for Regenerative Medicine, Graduate School of Medicine, Yamaguchi University, Ube, Yamaguchi 755‑8505, Japan, School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women's University, Nishinomiya, Hyogo 663‑8179, Japan, Department of Immunology, Graduate School of Medicine, Yamaguchi University, Ube, Yamaguchi 755‑8505, Japan
Published online on: August 1, 2021 https://doi.org/10.3892/ol.2021.12950
Article Number: 689
Copyright: © Samoto et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The interaction between prostate cancer cells and osteoblasts is essential for the development of bone metastasis. Previously, novel androgen receptor axis‑targeted agents (ARATs) were approved for metastatic castration‑naïve and non‑metastatic castration‑resistant prostate cancer (CRPC); both of which are pivotal for investigating the association between the bone microenvironment and tumors. The present study established a novel in vitro 3D microenvironment model that simulated the bone microenvironment of CRPC, and evaluated the drug susceptibility of ARATs and the efficacy of the combination of abiraterone and dutasteride. Green fluorescent protein‑transferred C4‑2 cells (a CRPC cell line) and red fluorescent protein‑transferred human osteoblasts differentiated from human mesenchymal stem cells were co‑cultured in chitosan nanofiber matrix‑coated culture plates to simulate the 3D scaffold of the bone microenvironment. The growth of C4‑2 was quantified using live‑cell imaging and the Cell3 iMager duos analysis system. The growth of C4‑2 colonies were quantified for a maximum of 30 days. The expression of TGF‑β increased and promoted EMT in C4‑2 cells co‑cultured with osteoblasts, indicating resistance to ARATs. The IC50 of each drug and the combination effect of abiraterone and dutasteride were evaluated using this model. Combination treatment with abiraterone and dutasteride synergistically inhibited the growth of C2‑4 colonies compared with individual investigational agents. This could be attributed to the reduction of 3‑keto‑5α‑abiraterone, an androgen receptor agonist. The bone microenvironment model of the present study is unique and useful for evaluating new drug susceptibility testing in prostate cancer cells. This model may help to reveal the unknown mechanisms underlying micro‑ to clinical bone metastasis in prostate cancer.

View Figures

View References

1	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
2	Hamdy FC, Donovan JL, Lane JA, Mason M, Metcalfe C, Holding P, Davis M, Peters TJ, Turner EL, Martin RM, et al: 10-Year outcomes after monitoring, surgery, or radiotherapy for localized prostate cancer. N Engl J Med. 375:1415–1424. 2016. View Article : Google Scholar : PubMed/NCBI
3	Weckermann D, Polzer B, Ragg T, Blana A, Schlimok G, Arnholdt H, Bertz S, Harzmann R and Klein CA: Perioperative activation of disseminated tumor cells in bone marrow of patients with prostate cancer. J Clin Oncol. 27:1549–1556. 2009. View Article : Google Scholar : PubMed/NCBI
4	National Cancer Institute, . SEER Cancer Stat Facts: Prostate Cancer. 2020.http://seer.cancer.gov/statfacts/html/prost.htmlNovember 14–2020
5	Sweeney CJ, Chen YH, Carducci M, Liu G, Jarrard DF, Eisenberger M, Wong YN, Hahn N, Kohli M, Cooney MM, et al: Chemohormonal therapy in metastatic hormone-sensitive prostate cancer. N Engl J Med. 373:737–746. 2015. View Article : Google Scholar : PubMed/NCBI
6	Wadosky KM and Koochekpour S: Molecular mechanisms underlying resistance to androgen deprivation therapy in prostate cancer. Oncotarget. 7:64447–64470. 2016. View Article : Google Scholar : PubMed/NCBI
7	Sumanasuriya S and Bono JD: Treatment of advanced prostate cancer-A review of current therapies and future promise. Cold Spring Harb Perspect Med. 8:a0306352018. View Article : Google Scholar : PubMed/NCBI
8	Pezaro C, Omlin A, Lorente D, Rodrigues DN, Ferraldeschi R, Bianchini D, Mukherji D, Riisnaes R, Altavilla A, Crespo M, et al: Visceral disease in castration-resistant prostate cancer. Eur Urol. 65:270–273. 2014. View Article : Google Scholar : PubMed/NCBI
9	Shiirevnyamba A, Takahashi T, Shan H, Ogawa H, Yano S, Kanayama H, Izumi K and Uehara H: Enhancement of osteoclastogenic activity in osteolytic prostate cancer cells by physical contact with osteoblasts. Br J Cancer. 104:505–513. 2011. View Article : Google Scholar : PubMed/NCBI
10	Antunes J, Gaspar VM, Ferreira L, Monteiro M, Henrique R, Jerónimo C and Mano JF: In-air production of 3D co-culture tumor spheroid hydrogels for expedited drug screening. Acta Biomater. 94:392–409. 2019. View Article : Google Scholar : PubMed/NCBI
11	Ryan CJ, Smith MR, Fizazi K, Saad F, Mulders PF, Sternberg CN, Miller K, Logothetis CJ, Shore ND, Small EJ, et al: Abiraterone acetate plus prednisone versus placebo plus prednisone in chemotherapy-naive men with metastatic castration-resistant prostate cancer (COU-AA-302): Final overall survival analysis of a randomised, double-blind, placebo-controlled phase 3 study. Lancet Oncol. 16:152–160. 2015. View Article : Google Scholar : PubMed/NCBI
12	Li Z, Alyamani M, Li J, Rogacki K, Abazeed M, Upadhyay SK, Balk SP, Taplin ME, Auchus RJ and Sharifi N: Redirecting abiraterone metabolism to fine-tune prostate cancer anti-androgen therapy. Nature. 533:547–551. 2016. View Article : Google Scholar : PubMed/NCBI
13	Andriole GL, Bostwick DG, Brawley OW, Gomella LG, Marberger M, Montorsi F, Pettaway CA, Tammela TL, Teloken C, Tindall DJ, et al: Effect of dutasteride on the risk of prostate cancer. N Engl J Med. 362:1192–1202. 2010. View Article : Google Scholar : PubMed/NCBI
14	Chou TC and Talalay P: Quantitative analysis of dose-effect relationships: The combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul. 22:27–55. 1984. View Article : Google Scholar : PubMed/NCBI
15	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
16	Matsuyama H, Shiota M, Tashiro K, Kanji H, Horiyama S, Eto M, Egawa S, Haginaka J and Inoue R: Phase II study of the efficacy of abiraterone acetate with dutasteride for castration resistant prostate cancer. J Clin Oncol. 39:112. 2021. View Article : Google Scholar
17	Horoszewicz JS, Leong SS, Kawinski E, Karr JP, Rosenthal H, Chu TM, Mirand EA and Murphy GP: LNCaP model of human prostatic carcinoma. Cancer Res. 43:1809–1818. 1983.PubMed/NCBI
18	Cai C, Chen S, Ng P, Bubley GJ, Nelson PS, Mostaghel EA, Marck B, Matsumoto AM, Simon NI, Wang H, et al: Intratumoral de novo steroid synthesis activates androgen receptor in castration-resistant prostate cancer and is upregulated by treatment with CYP17A1 inhibitors. Cancer Res. 71:6503–6513. 2011. View Article : Google Scholar : PubMed/NCBI
19	Rajendran D, Hussain A, Yip D, Parekh A, Shrirao A and Cho CH: Long-term liver-specific functions of hepatocytes in electrospun chitosan nanofiber scaffolds coated with fibronectin. J Biomed Mater Res A. 105:2119–2128. 2017. View Article : Google Scholar : PubMed/NCBI
20	Hussain A, Collins G, Yip D and Cho CH: Functional 3-D cardiac co-culture model using bioactive chitosan nanofiber scaffolds. Biotechnol Bioeng. 110:637–647. 2013. View Article : Google Scholar : PubMed/NCBI
21	Ho MH, Liao MH, Lin YL, Lai CH, Lin PI and Chen RM: Improving effects of chitosan nanofiber scaffolds on osteoblast proliferation and maturation. Int J Nanomedicine. 9:4293–4304. 2014.PubMed/NCBI
22	Kimura Y, Matsugaki A, Sekita A and Nakano T: Alteration of osteoblast arrangement via direct attack by cancer cells: New insights into bone metastasis. Sci Rep. 7:448242017. View Article : Google Scholar : PubMed/NCBI
23	Kingsley LA, Fournier PG, Chirgwin JM and Guise TA: Molecular biology of bone metastasis. Mol Cancer Ther. 6:2609–2617. 2007. View Article : Google Scholar : PubMed/NCBI
24	Wang N, Docherty FE, Brown HK, Reeves KJ, Fowles AC, Ottewell PD, Dear TN, Holen I, Croucher PI and Eaton CL: Prostate cancer cells preferentially home to osteoblast-rich areas in the early stages of bone metastasis: Evidence from in vivo models. J Bone Miner Res. 29:2688–2696. 2014. View Article : Google Scholar : PubMed/NCBI
25	Klein CA: Selection and adaptation during metastatic cancer progression. Nature. 501:365–372. 2013. View Article : Google Scholar : PubMed/NCBI
26	Guise TA, Mohammad KS, Clines G, Stebbins EG, Wong DH, Higgins LS, Vessella R, Corey E, Padalecki S, Suva L and Chirgwin JM: Basic mechanisms responsible for osteolytic and osteoblastic bone metastases. Clin Cancer Res. 12 (Suppl 1):6213S–6216S. 2006. View Article : Google Scholar : PubMed/NCBI
27	Weilbaecher KN, Guise TA and McCauley LK: Cancer to bone: A fatal attraction. Nat Rev Cancer. 11:411–425. 2011. View Article : Google Scholar : PubMed/NCBI
28	Chiechi A, Waning DL, Stayrook KR, Buijs JT, Guise TA and Mohammad KS: Role of TGF-β in breast cancer bone metastases. Adv Biosci Biotechnol. 4:15–30. 2013. View Article : Google Scholar : PubMed/NCBI
29	Pickup M, Novitskiy S and Moses HL: The roles of TGFβ in the tumour microenvironment. Nat Rev Cancer. 13:788–799. 2013. View Article : Google Scholar : PubMed/NCBI
30	Juárez P and Guise TA: TGF-β in cancer and bone: Implications for treatment of bone metastases. Bone. 48:23–29. 2011. View Article : Google Scholar : PubMed/NCBI
31	Montanari M, Rossetti S, Cavaliere C, D'Aniello C, Malzone MG, Vanacore D, Di Franco R, La Mantia E, Iovane G, Piscitelli R, et al: Epithelial-mesenchymal transition in prostate cancer: An overview. Oncotarget. 8:35376–35389. 2017. View Article : Google Scholar : PubMed/NCBI
32	Nakazawa M and Kyprianou N: Epithelial-mesenchymal-transition regulators in prostate cancer: Androgens and beyond. J Steroid Biochem Mol Biol. 166:84–90. 2017. View Article : Google Scholar : PubMed/NCBI
33	Shiota M, Itsumi M, Takeuchi A, Imada K, Yokomizo A, Kuruma H, Inokuchi J, Tatsugami K, Uchiumi T, Oda Y and Naito S: Crosstalk between epithelial-mesenchymal transition and castration resistance mediated by Twist1/AR signaling in prostate cancer. Endocr Relat Cancer. 22:889–900. 2015. View Article : Google Scholar : PubMed/NCBI
34	Fizazi K, Yang J, Peleg S, Sikes CR, Kreimann EL, Daliani D, Olive M, Raymond KA, Janus TJ, Logothetis CJ, et al: Prostate cancer cells-osteoblast interaction shifts expression of growth/survival-related genes in prostate cancer and reduces expression of osteoprotegerin in osteoblasts. Clin Cancer Res. 9:2587–2597. 2003.PubMed/NCBI
35	Xu K, Wang Z, Copland JA, Chakrabarti R and Florczyk SJ: 3D porous chitosan-chondroitin sulfate scaffolds promote epithelial to mesenchymal transition in prostate cancer cells. Biomaterials. 254:1201262020. View Article : Google Scholar : PubMed/NCBI
36	Xu K, Ganapathy K, Andl T, Wang Z, Copland JA, Chakrabarti R and Florczyk SJ: 3D porous chitosan-alginate scaffold stiffness promotes differential responses in prostate cancer cell lines. Biomaterials. 217:1193112019. View Article : Google Scholar : PubMed/NCBI
37	Wang K, Kievit FM, Florczyk SJ, Stephen ZR and Zhang M: 3D porous chitosan-alginate scaffolds as an in vitro model for evaluating nanoparticle-mediated tumor targeting and gene delivery to prostate cancer. Biomacromolecules. 16:3362–3372. 2015. View Article : Google Scholar : PubMed/NCBI
38	Elbadawy M, Abugomaa A, Yamawaki H, Usui T and Sasaki K: Development of prostate cancer organoid culture models in basic medicine and translational research. Cancers (Basel). 12:7772020. View Article : Google Scholar : PubMed/NCBI
39	Bartucci M, Ferrari AC, Kim IY, Ploss A, Yarmush M and Sabaawy HE: Personalized medicine approaches in prostate cancer employing patient derived 3D organoids and humanized mice. Front Cell Dev Biol. 4:642016. View Article : Google Scholar : PubMed/NCBI
40	Kuchimaru T, Kataoka N, Nakagawa K, Isozaki T, Miyabara H, Minegishi M, Kadonosono T and Kizaka-Kondoh S: A reliable murine model of bone metastasis by injecting cancer cells through caudal arteries. Nat Commun. 30:29812018. View Article : Google Scholar : PubMed/NCBI