<em>In ovo</em> chorioallantoic membrane assay as a xenograft model for pediatric rhabdomyosarcoma

Shoji,Chika; Kikuchi,Ken; Yoshida,Hideki; Miyachi,Mitsuru; Yagyu,Shigeki; Tsuchiya,Kunihiko; Nakaya,Takaaki; Hosoi,Hajime; Iehara,Tomoko

doi:10.3892/or.2023.8513

In ovo chorioallantoic membrane assay as a xenograft model for pediatric rhabdomyosarcoma

Authors:
- Chika Shoji
- Ken Kikuchi
- Hideki Yoshida
- Mitsuru Miyachi
- Shigeki Yagyu
- Kunihiko Tsuchiya
- Takaaki Nakaya
- Hajime Hosoi
- Tomoko Iehara
View Affiliations

Affiliations: Department of Pediatrics, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602‑8566, Japan, Department of Infectious Diseases, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602‑8566, Japan
Published online on: March 2, 2023 https://doi.org/10.3892/or.2023.8513
Article Number: 76
Copyright: © Shoji 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

Rhabdomyosarcoma (RMS) is the most common highly malignant pediatric soft tissue sarcoma. While recent multidisciplinary treatments have improved the 5‑year survival rate of low/intermediate‑risk patients to 70‑90%, there are various complications that arise due to treatment‑related toxicities. Immunodeficient mice‑derived xenograft models have been widely used in cancer drug research; however, these models have some limitations, including i) they are time‑consuming and expensive, ii) their use needs to be approved by animal experimental ethics committees, and iii) the inability to visualize where tumor cells or tissues were engrafted. The present study performed a chorioallantoic membrane (CAM) assay in fertilized chicken eggs, which is time‑saving, simple, and easy to standardize and handle because of the high vascularization and the immature immune system of the fertilized eggs. The present study aimed to examine the usability of the CAM assay as a novel therapeutic model for the development of precision medicine for pediatric cancer. A protocol was developed for constructing cell line‑derived xenograft (CDX) models using a CAM assay by transplanting RMS cells on the CAM. It was then examined as to whether these CDX models could be used as therapeutic drug evaluation models using vincristine (VCR) and human RMS cell lines. After grafting and culturing the RMS cell suspension on the CAM, three‑dimensional proliferation over time was observed visually and by comparing volumes. VCR reduced the size of the RMS tumor on the CAM in a dose‑dependent manner. Currently, treatment strategies based on patient‑specific oncogenic backgrounds have not been adequately developed in the field of pediatric cancer. The establishment of a CDX model with the CAM assay may lead to the advancement of precision medicine and help formulate novel therapeutic strategies for intractable pediatric cancer.

View Figures

View References

1	Do K, O'Sullivan Coyne G and Chen AP: An overview of the NCI precision medicine trials-NCI MATCH and MPACT. Chin Clin Oncol. 4:312015.PubMed/NCBI
2	Lokman NA, Elder ASF, Ricciardelli C and Oehler MK: Chick chorioallantoic membrane (CAM) assay as an in vivo model to study the effect of newly identified molecules on ovarian cancer invasion and metastasis. Int J Mol Sci. 13:9959–9970. 2012. View Article : Google Scholar : PubMed/NCBI
3	DeBord LC, Pathak RR, Villaneuva M, Liu HC, Harrington DA, Yu W, Lewis MT and Sikora AG: The chick chorioallantoic membrane (CAM) as a versatile patient-derived xenograft (PDX) platform for precision medicine and preclinical research. Am J Cancer Res. 8:1642–1660. 2018.PubMed/NCBI
4	Chu PY, Koh AP, Antony J and Huang RY: Applications of the chick chorioallantoic membrane as an alternative model for cancer studies. Cells Tissues Organs. 211:222–237. 2022. View Article : Google Scholar : PubMed/NCBI
5	Valdes TI, Kreutzer D and Moussy F: The chick chorioallantoic membrane as a novel in vivo model for the testing of biomaterials. J Biomed Mater Res. 62:273–282. 2002. View Article : Google Scholar : PubMed/NCBI
6	Ribatti D: Chicken chorioallantoic membrane angiogenesis model. Methods Mol Biol. 843:47–57. 2012. View Article : Google Scholar : PubMed/NCBI
7	Fiorentzis M, Viestenz A, Siebolts U, Seitz B, Coupland SE and Heinzelmann J: The potential use of electrochemotherapy in the treatment of uveal melanoma: In vitro results in 3D tumor cultures and in vivo results in a chick embryo model. Cancers (Basel). 11:13442019. View Article : Google Scholar : PubMed/NCBI
8	Moreno-Jiménez I, Lanham SA, Kanczler JM, Hulsart-Billstrom G, Evans ND and Oreffo ROC: Remodelling of human bone on the chorioallantoic membrane of the chicken egg: De novo bone formation and resorption. J Tissue Eng Regen Med. 12:1877–1890. 2018. View Article : Google Scholar : PubMed/NCBI
9	Rasmussen SV, Berlow NE, Price LH, Mansoor A, Cairo S, Rugonyi S and Keller C: Preclinical therapeutics ex ovo quail eggs as a biomimetic automation-ready xenograft platform. Sci Rep. 11:233022021. View Article : Google Scholar : PubMed/NCBI
10	Kunz P, Schenker A, Sähr H, Lehner B and Fellenberg J: Optimization of the chicken chorioallantoic membrane assay as reliable in vivo model for the analysis of osteosarcoma. PLoS One. 14:e02153122019. View Article : Google Scholar : PubMed/NCBI
11	Ribatti D: The chick embryo chorioallantoic membrane (CAM) assay. Reprod Toxicol. 70:97–101. 2017. View Article : Google Scholar : PubMed/NCBI
12	Nowak-Sliwinska P, Segura T and Iruela-Arispe ML: The chicken chorioallantoic membrane model in biology, medicine and bioengineering. Angiogenesis. 17:779–804. 2014. View Article : Google Scholar : PubMed/NCBI
13	Vu BT, Shahin SA, Croissant J, Fatieiev Y, Matsumoto K, Le-Hoang Doan T, Yik T, Simargi S, Conteras A, Ratliff L, et al: Chick chorioallantoic membrane assay as an in vivo model to study the effect of nanoparticle-based anticancer drugs in ovarian cancer. Sci Rep. 8:85242018. View Article : Google Scholar : PubMed/NCBI
14	Dasgupta R, Fuchs J and Rodeberg D: Rhabdomyosarcoma. Semin Pediatr Surg. 25:276–283. 2016. View Article : Google Scholar : PubMed/NCBI
15	Ognjanovic S, Linabery AM, Charbonneau B and Ross JA: Trends in childhood rhabdomyosarcoma incidence and survival in the United States, 1975–2005. Cancer. 115:4218–4226. 2009. View Article : Google Scholar : PubMed/NCBI
16	Nakagawa N, Kikuchi K, Yagyu S, Miyachi M, Iehara T, Tajiri T, Sakai T and Hosoi H: Mutations in the RAS pathway as potential precision medicine targets in treatment of rhabdomyosarcoma. Biochem Biophys Res Commun. 512:524–530. 2019. View Article : Google Scholar : PubMed/NCBI
17	Ouchi K, Miyachi M, Yagyu S, Kikuchi K, Kuwahara Y, Tsuchiya K, Iehara T and Hosoi H: Oncogenic role of HMGA2 in fusion-negative rhabdomyosarcoma cells. Cancer Cell Int. 20:1922020. View Article : Google Scholar : PubMed/NCBI
18	Gurria JP and Dasgupta R: Rhabdomyosarcoma and extraosseous ewing sarcoma. Children (Basel). 5:1652018.PubMed/NCBI
19	Malempati S and Hawkins DS: Rhabdomyosarcoma: Review of the children's oncology group (COG) soft-tissue sarcoma committee experience and rationale for current COG studies. Pediatr Blood Cancer. 59:5–10. 2012. View Article : Google Scholar : PubMed/NCBI
20	Otabe O, Kikuchi K, Tsuchiya K, Katsumi Y, Yagyu S, Miyachi M, Iehara T and Hosoi H: MET/ERK2 pathway regulates the motility of human alveolar rhabdomyosarcoma cells. Oncol Rep. 37:98–104. 2017. View Article : Google Scholar : PubMed/NCBI
21	Davicioni E, Anderson MJ, Finckenstein FG, Lynch JC, Qualman SJ, Shimada H, Schofield DE, Buckley JD, Meyer WH, Sorensen PH and Triche TJ: Molecular classification of rhabdomyosarcoma-genotypic and phenotypic determinants of diagnosis: A report from the children's oncology group. Am J Pathol. 174:550–564. 2009. View Article : Google Scholar : PubMed/NCBI
22	Williamson D, Missiaglia E, de Reyniès A, Pierron G, Thuille B, Palenzuela G, Thway K, Orbach D, Laé M, Fréneaux P, et al: Fusion gene-negative alveolar rhabdomyosarcoma is clinically and molecularly indistinguishable from embryonal rhabdomyosarcoma. J Clin Oncol. 28:2151–2158. 2010. View Article : Google Scholar : PubMed/NCBI
23	Meza JL, Anderson J, Pappo AS and Meyer WH; Children's Oncology Group, : Analysis of prognostic factors in patients with nonmetastatic rhabdomyosarcoma treated on intergroup rhabdomyosarcoma studies III and IV: The children's oncology group. J Clin Oncol. 24:3844–3851. 2006. View Article : Google Scholar : PubMed/NCBI
24	Arndt CAS, Stoner JA, Hawkins DS, Rodeberg DA, Hayes-Jordan AA, Paidas CN, Parham DM, Teot LA, Wharam MD, Breneman JC, et al: Vincristine, actinomycin, and cyclophosphamide compared with vincristine, actinomycin, and cyclophosphamide alternating with vincristine, topotecan, and cyclophosphamide for intermediate-risk rhabdomyosarcoma: Children's oncology group study D9803. J Clin Oncol. 27:5182–5188. 2009. View Article : Google Scholar : PubMed/NCBI
25	Oberlin O, Rey A, Lyden E, Bisogno G, Stevens MC, Meyer WH, Carli M and Anderson JR: Prognostic factors in metastatic rhabdomyosarcomas: Results of a pooled analysis from United States and European cooperative groups. J Clin Oncol. 26:2384–2389. 2008. View Article : Google Scholar : PubMed/NCBI
26	Arndt CAS, Rose PS, Folpe AL and Laack NN: Common musculoskeletal tumors of childhood and adolescence. Mayo Clin Proc. 87:475–487. 2012. View Article : Google Scholar : PubMed/NCBI
27	Lockney NA, Friedman DN, Wexler LH, Sklar CA, Casey DL and Wolden SL: Late toxicities of intensity-modulated radiation therapy for head and neck rhabdomyosarcoma. Pediatr Blood Cancer. 63:1608–1614. 2016. View Article : Google Scholar : PubMed/NCBI
28	Clement SC, Schoot RA, Slater O, Chisholm JC, Abela C, Balm AJM, van den Brekel MW, Breunis WB, Chang YC, Davila Fajardo R, et al: Endocrine disorders among long-term survivors of childhood head and neck rhabdomyosarcoma. Eur J Cancer. 54:1–10. 2016. View Article : Google Scholar : PubMed/NCBI
29	Walterhouse D and Watson A: Optimal management strategies for rhabdomyosarcoma in children. Paediatr Drugs. 9:391–400. 2007. View Article : Google Scholar : PubMed/NCBI
30	Douglass EC, Valentine M, Etcubanas E, Parham D, Webber BL, Houghton PJ, Houghton JA and Green AA: A specific chromosomal abnormality in rhabdomyosarcoma. Cytogenet Cell Genet. 45:148–155. 1987. View Article : Google Scholar : PubMed/NCBI
31	Kanda Y: Investigation of the freely available easy-to-use software ‘EZR’ for medical statistics. Bone Marrow Transplant. 48:452–458. 2013. View Article : Google Scholar : PubMed/NCBI
32	Schwartz SO and Stansbury F: Significance of nucleated red blood cells in peripheral blood; analysis of 1,496 cases. J Am Med Assoc. 154:1339–1340. 1954. View Article : Google Scholar : PubMed/NCBI
33	Ghaffari-Tabrizi-Wizsy N, Passegger CA, Nebel L, Krismer F, Herzer-Schneidhofer G, Schwach G and Pfragner R: The avian chorioallantoic membrane as an alternative tool to study medullary thyroid cancer. Endocr Connect. 8:462–467. 2019. View Article : Google Scholar : PubMed/NCBI
34	Linardic CM, Downie DL, Qualman S, Bentley RC and Counter CM: Genetic modeling of human rhabdomyosarcoma. Cancer Res. 65:4490–4495. 2005. View Article : Google Scholar : PubMed/NCBI
35	Dolgikh N, Hugle M, Vogler M and Fulda S: NRAS-mutated rhabdomyosarcoma cells are vulnerable to mitochondrial apoptosis induced by coinhibition of MEK and PI3Kα. Cancer Res. 78:2000–2013. 2018. View Article : Google Scholar : PubMed/NCBI
36	Heinicke U, Kupka J, Fichter I and Fulda S: Critical role of mitochondria-mediated apoptosis for JNJ-26481585-induced antitumor activity in rhabdomyosarcoma. Oncogene. 35:3729–3741. 2016. View Article : Google Scholar : PubMed/NCBI
37	Asam C, Buerger K, Felthaus O, Brébant V, Rachel R, Prantl L, Witzgall R, Haerteis S and Aung T: Subcellular localization of the chemotherapeutic agent doxorubicin in renal epithelial cells and in tumor cells using correlative light and electron microscopy. Clin Hemorheol Microcirc. 73:157–167. 2019. View Article : Google Scholar : PubMed/NCBI
38	Abraham J, Prajapati SI, Nishijo K, Schaffer BS, Taniguchi E, Kilcoyne A, McCleish AT, Nelon LD, Giles FG, Efstratiadis A, et al: Evasion mechanisms to Igf1r inhibition in rhabdomyosarcoma. Mol Cancer Ther. 10:697–707. 2011. View Article : Google Scholar : PubMed/NCBI