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Journal of Experimental & Clinical Cancer Research
Published in: Journal of Experimental & Clinical Cancer Research 1/2021

Open Access 01-12-2021 | NSCLC | Research

Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

Authors: Ruo-Yue Fan, Jia-Qi Wu, Yu-Yang Liu, Xiang-Yu Liu, Si-Tong Qian, Chong-Yong Li, Ping Wei, Zhe Song, Ming-Fang He

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2021

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Abstract

Background

Brain metastasis (BM) is thought to be related to the mortality and poor prognosis of non-small cell lung cancer (NSCLC). Despite promising development of NSCLC treatment, the treatment of NSCLC BM is still not optimistic due to the existence of the blood-brain barrier (BBB) that prevent drug penetration, as well as the short median survival time of the patients left for treatment. In this context, further development of quick and effective pre-clinical models is needed in NSCLC BM treatment. Here, we report a model system using zebrafish to promote the development of drugs for patients with NSCLC BM.

Methods

Three different NSCLC cell lines (H1975, A549 and H1299) were used to establish zebrafish BM models. The embryo age and cell number for injection were first optimized. Metastatic cells were observed in the brain blood vessels of zebrafish and were verified by hematoxylin-eosin (HE) staining. Then, the metastasis potentials of H1975 and A549 with manipulated microRNA-330-3p (miR-330-3p) expression were also investigated. Finally, sensitivities of H1975 and A549 to osimertinib and gefitinib were tested.

Results

This zebrafish BM model could distinguish NSCLC cell lines with different BM potential. Over-expressed miR-330-p significantly improved the BM potential of the A549 cells while knockdown miR-330-p reduced the BM ability of the H1975 cells. Both osimertinib and gefitinib showed inhibition effect in zebrafish BM model with the inhibition rate higher than 50 %. H1975 cell showed much higher sensitivity to osimertinib rather than gefitinib both in vivo and in vitro.

Conclusions

We established zebrafish brain metastasis model for studying mechanism and treatment of NSCLC BM. This study provided a useful model for NSCLC brain metastasis that could be used to study the mechanism that drive NSCLC cells to the brain as well as identify potential therapeutic options.
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Literature
1.
2.
Mehta MP, Rodrigus P, Terhaard CH, Rao A, Suh J, Roa W, et al. Survival and neurologic outcomes in a randomized trial of motexafin gadolinium and whole-brain radiation therapy in brain metastases. J Clin Oncol. 2003;21(13):2529–36.PubMedCrossRef
3.
Nayak L, Lee EQ, Wen PY. Epidemiology of brain metastases. Current oncology reports. 2012;14(1):48–54.PubMedCrossRef
4.
An N, Jing W, Wang H, Li J, Liu Y, Yu J, et al. Risk factors for brain metastases in patients with non-small-cell lung cancer. Cancer medicine. 2018;7(12):6357–64.PubMedPubMedCentralCrossRef
5.
Hung YP, Sholl LM. Diagnostic and Predictive Immunohistochemistry for Non-Small Cell Lung Carcinomas. Adv Anat Pathol. 2018;25(6):374–86.PubMedCrossRef
6.
Niu Z, Guo S, Cao J, Zhang Y, Guo X, Grossi F, et al. Immune checkpoint inhibitors for treatment of small-cell lung cancer: a systematic review and meta-analysis. Ann Transl Med. 2021;9(8):705.PubMedPubMedCentralCrossRef
7.
Li W, Yu H. Separating or combining immune checkpoint inhibitors (ICIs) and radiotherapy in the treatment of NSCLC brain metastases. Journal of cancer research and clinical oncology. 2020;146(1):137–52.PubMedCrossRef
8.
Alomari A, Rauch PJ, Orsaria M, Minja FJ, Chiang VL, Vortmeyer AO. Radiologic and histologic consequences of radiosurgery for brain tumors. Journal of neuro-oncology. 2014;117(1):33–42.PubMedCrossRef
9.
Khan AJ, Dicker AP. On the merits and limitations of whole-brain radiation therapy. J Clin Oncol. 2013;31(1):11–3.PubMedCrossRef
10.
Duma N, Santana-Davila R, Molina JR, Herbst RS, Morgensztern D, Boshoff C, et al. Non-Small Cell Lung Cancer: Epidemiology, Screening, Diagnosis, and Treatment. Mayo Clin Proc. 2019;94(8):1623–40.PubMedCrossRef
11.
Herbst RS, Morgensztern D, Boshoff C. The biology and management of non-small cell lung cancer. Nature. 2018;553(7689):446–54.PubMedCrossRef
12.
Ko EC, Raben D, Formenti SC. The Integration of Radiotherapy with Immunotherapy for the Treatment of Non-Small Cell Lung Cancer. Clin Cancer Res. 2018;24(23):5792–806.PubMedCrossRef
13.
Shetty V, Babu S. Management of CNS metastases in patients with EGFR mutation-positive NSCLC. Indian J Cancer. 2019;56(Supplement):S31-S7.
14.
Wittlinger F, Laufer SA. The pre-clinical discovery and development of osimertinib used to treat non-small cell lung cancer. Expert opinion on drug discovery. 2021.
15.
Cai XT, Miao J, Sun RW, Wang SN, Molina-Vila MA, Chaib I, et al. Dihydroartemisinin overcomes the resistance to osimertinib in EGFR-mutant non-small-cell lung cancer. Pharmacol Res. 2021;170:105701.
16.
Wang S, Hu C, Xie F, Liu Y. Use of Programmed Death Receptor-1 and/or Programmed Death Ligand 1 Inhibitors for the Treatment of Brain Metastasis of Lung Cancer. OncoTargets and therapy. 2020;13:667–83.PubMedPubMedCentralCrossRef
17.
MacRae CA, Peterson RT. Zebrafish as tools for drug discovery. Nature reviews Drug discovery. 2015;14(10):721–31.PubMedCrossRef
18.
Brown HK, Schiavone K, Tazzyman S, Heymann D, Chico TJ. Zebrafish xenograft models of cancer and metastasis for drug discovery. Expert opinion on drug discovery. 2017;12(4):379–89.PubMedCrossRef
19.
Völkel P, Dupret B, Le Bourhis X, Angrand PO. [The zebrafish model in oncology]. Medecine sciences: M/S. 2018;34(4):345-53.
20.
Wu JQ, Zhai J, Li CY, Tan AM, Wei P, Shen LZ, et al. Patient-derived xenograft in zebrafish embryos: a new platform for translational research in gastric cancer. J Exp Clin Cancer Res. 2017;36(1):160.PubMedPubMedCentralCrossRef
21.
Fior R, Póvoa V, Mendes RV, Carvalho T, Gomes A, Figueiredo N, et al. Single-cell functional and chemosensitive profiling of combinatorial colorectal therapy in zebrafish xenografts. Proc Natl Acad Sci U S A. 2017;114(39):E8234-E43.CrossRef
22.
Zeng A, Ye T, Cao D, Huang X, Yang Y, Chen X, et al. Identify a Blood-Brain Barrier Penetrating Drug-TNB using Zebrafish Orthotopic Glioblastoma Xenograft Model. Scientific reports. 2017;7(1):14372.PubMedPubMedCentralCrossRef
23.
Li Y, Chen T, Miao X, Yi X, Wang X, Zhao H, et al. Zebrafish: A promising in vivo model for assessing the delivery of natural products, fluorescence dyes and drugs across the blood-brain barrier. Pharmacol Res. 2017;125(Pt B):246–57.PubMedCrossRef
24.
Chen L, Groenewoud A, Tulotta C, Zoni E, Kruithof-de Julio M, van der Horst G, et al. A zebrafish xenograft model for studying human cancer stem cells in distant metastasis and therapy response. Methods Cell Biol. 2017;138:471–96.PubMedCrossRef
25.
Kim EA, Lee JH, Heo SJ, Jeon YJ. Saringosterol acetate isolated from Hizikia fusiforme, an edible brown alga, suppressed hepatocellular carcinoma growth and metastasis in a zebrafish xenograft model. Chemico-biological interactions. 2021;335:109362.PubMedCrossRef
26.
Nakayama J, Lu JW, Makinoshima H, Gong Z. A Novel Zebrafish Model of Metastasis Identifies the HSD11β1 Inhibitor Adrenosterone as a Suppressor of Epithelial-Mesenchymal Transition and Metastatic Dissemination. Molecular cancer research: MCR. 2020;18(3):477–87.PubMedCrossRef
27.
Lee HW, Lee JI, Lee SJ, Cho HJ, Song HJ, Jeong DE, et al. Patient-derived xenografts from non-small cell lung cancer brain metastases are valuable translational platforms for the development of personalized targeted therapy. Clin Cancer Res. 2015;21(5):1172–82.PubMedCrossRef
28.
Yousefi M, Bahrami T, Salmaninejad A, Nosrati R, Ghaffari P, Ghaffari SH. Lung cancer-associated brain metastasis: Molecular mechanisms and therapeutic options. Cell Oncol (Dordr). 2017;40(5):419–41.CrossRef
29.
Quiñonez-Silvero C, Hübner K, Herzog W. Development of the brain vasculature and the blood-brain barrier in zebrafish. Dev Biol. 2020;457(2):181–90.PubMedCrossRef
30.
Eliceiri BP, Gonzalez AM, Baird A. Zebrafish model of the blood-brain barrier: morphological and permeability studies. Methods in molecular biology (Clifton, NJ). 2011;686:371–8.
31.
Avenel C, Tolf A, Dragomir A, Carlbom IB. Glandular Segmentation of Prostate Cancer: An Illustration of How the Choice of Histopathological Stain Is One Key to Success for Computational Pathology. Frontiers in bioengineering and biotechnology. 2019;7:125.PubMedPubMedCentralCrossRef
32.
Guo Y, Fan Y, Pei X. Fangjihuangqi Decoction inhibits MDA-MB-231 cell invasion in vitro and decreases tumor growth and metastasis in triple-negative breast cancer xenografts tumor zebrafish model. Cancer medicine. 2020;9(7):2564–78.PubMedPubMedCentralCrossRef
33.
Razak NA, Abu N, Ho WY, Zamberi NR, Tan SW, Alitheen NB, et al. Cytotoxicity of eupatorin in MCF-7 and MDA-MB-231 human breast cancer cells via cell cycle arrest, anti-angiogenesis and induction of apoptosis. Scientific reports. 2019;9(1):1514.PubMedPubMedCentralCrossRef
34.
Li Y, Zhu X, Xu W, Wang D, Yan J. miR-330 regulates the proliferation of colorectal cancer cells by targeting Cdc42. Biochemical and biophysical research communications. 2013;431(3):560–5.PubMedCrossRef
35.
Meng H, Wang K, Chen X, Guan X, Hu L, Xiong G, et al. MicroRNA-330-3p functions as an oncogene in human esophageal cancer by targeting programmed cell death 4. American journal of cancer research. 2015;5(3):1062–75.PubMedPubMedCentral
36.
Wei CH, Wu G, Cai Q, Gao XC, Tong F, Zhou R, et al. MicroRNA-330-3p promotes cell invasion and metastasis in non-small cell lung cancer through GRIA3 by activating MAPK/ERK signaling pathway. J Hematol Oncol. 2017;10(1):125.PubMedPubMedCentralCrossRef
37.
Wei C, Zhang R, Cai Q, Gao X, Tong F, Dong J, et al. MicroRNA-330-3p promotes brain metastasis and epithelial-mesenchymal transition via GRIA3 in non-small cell lung cancer. Aging. 2019;11(17):6734–61.PubMedPubMedCentralCrossRef
38.
Lim ZF, Ma PC. Emerging insights of tumor heterogeneity and drug resistance mechanisms in lung cancer targeted therapy. J Hematol Oncol. 2019;12(1):134.PubMedPubMedCentralCrossRef
39.
40.
Hotz JM, Thomas JR, Katz EN, Robey RW, Horibata S, Gottesman MM. ATP-binding cassette transporters at the zebrafish blood-brain barrier and the potential utility of the zebrafish as an in vivo model. Cancer drug resistance (Alhambra, Calif). 2021;4:620–33.
41.
O’Brown NM, Megason SG, Gu C. Suppression of transcytosis regulates zebrafish blood-brain barrier function. Elife. 2019;8:e47326.
42.
Póvoa V, Rebelo de Almeida C, Maia-Gil M, Sobral D, Domingues M, Martinez-Lopez M, et al. Innate immune evasion revealed in a colorectal zebrafish xenograft model. Nature communications. 2021;12(1):1156.PubMedPubMedCentralCrossRef
43.
Bajoghli B, Dick AM, Claasen A, Doll L, Aghaallaei N. Zebrafish and Medaka: Two Teleost Models of T-Cell and Thymic Development. Int J Mol Sci. 2019;20(17):4179.
44.
45.
Schuermann A, Helker CS, Herzog W. Angiogenesis in zebrafish. Seminars in cell & developmental biology. 2014;31:106–14.CrossRef
46.
Zhang Q, Lu S, Li T, Yu L, Zhang Y, Zeng H, et al. ACE2 inhibits breast cancer angiogenesis via suppressing the VEGFa/VEGFR2/ERK pathway. J Exp Clin Cancer Res. 2019;38(1):173.PubMedPubMedCentralCrossRef
47.
Huang JQ, Wei FK, Xu XL, Ye SX, Song JW, Ding PK, et al. SOX9 drives the epithelial-mesenchymal transition in non-small-cell lung cancer through the Wnt/β-catenin pathway. Journal of translational medicine. 2019;17(1):143.PubMedPubMedCentralCrossRef
48.
Kim J, Chuang HC, Wolf NK, Nicolai CJ, Raulet DH, Saijo K, et al. Tumor-induced disruption of the blood-brain barrier promotes host death. Dev cell. 2021;56(19):2712–21.
49.
Yonemori K, Tsuta K, Ono M, Shimizu C, Hirakawa A, Hasegawa T, et al. Disruption of the blood brain barrier by brain metastases of triple-negative and basal-type breast cancer but not HER2/neu-positive breast cancer. Cancer. 2010;116(2):302–8.PubMedCrossRef
50.
Mehta AI, Brufsky AM, Sampson JH. Therapeutic approaches for HER2-positive brain metastases: circumventing the blood-brain barrier. Cancer treatment reviews. 2013;39(3):261–9.PubMedCrossRef
51.
Astell KR, Sieger D. Investigating microglia-brain tumor cell interactions in vivo in the larval zebrafish brain. Methods Cell Biol. 2017;138:593–626.PubMedCrossRef
52.
Ballard P, Yates JW, Yang Z, Kim DW, Yang JC, Cantarini M, et al. Preclinical Comparison of Osimertinib with Other EGFR-TKIs in EGFR-Mutant NSCLC Brain Metastases Models, and Early Evidence of Clinical Brain Metastases Activity. Clin Cancer Res. 2016;22(20):5130–40.PubMedCrossRef
53.
Arora S, Ranade AR, Tran NL, Nasser S, Sridhar S, Korn RL, et al. MicroRNA-328 is associated with (non-small) cell lung cancer (NSCLC) brain metastasis and mediates NSCLC migration. International journal of cancer. 2011;129(11):2621–31.PubMedPubMedCentralCrossRef
54.
Lu HY, Su D, Pan XD, Jiang H, Ma SL. Mutation and expression of multiple treatment response-related genes in a population with locally advanced non-small cell lung cancer. Oncology letters. 2012;3(2):415–20.PubMedCrossRef
55.
Novoa B, Figueras A. Zebrafish: model for the study of inflammation and the innate immune response to infectious diseases. Adv Exp Med Biol. 2012;946:253–75.PubMedCrossRef
56.
Usai A, Di Franco G, Colucci P, Pollina LE, Vasile E, Funel N, et al. A Model of a Zebrafish Avatar for Co-Clinical Trials. Cancers. 2020;12(3):677.
57.
Di Franco G, Usai A, Funel N, Palmeri M, Montesanti IER, Bianchini M, et al. Use of zebrafish embryos as avatar of patients with pancreatic cancer: A new xenotransplantation model towards personalized medicine. World journal of gastroenterology. 2020;26(21):2792–809.PubMedPubMedCentralCrossRef
58.
Fazio M, Ablain J, Chuan Y, Langenau DM, Zon LI. Zebrafish patient avatars in cancer biology and precision cancer therapy. Nature reviews Cancer. 2020;20(5):263–73.PubMedPubMedCentralCrossRef
59.
Miserocchi G, Mercatali L, Liverani C, De Vita A, Spadazzi C, Pieri F, et al. Management and potentialities of primary cancer cultures in preclinical and translational studies. Journal of translational medicine. 2017;15(1):229.PubMedPubMedCentralCrossRef
Metadata
Title
Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis
Authors
Ruo-Yue Fan
Jia-Qi Wu
Yu-Yang Liu
Xiang-Yu Liu
Si-Tong Qian
Chong-Yong Li
Ping Wei
Zhe Song
Ming-Fang He
Publication date
01-12-2021
Publisher
BioMed Central
Published in
Journal of Experimental & Clinical Cancer Research / Issue 1/2021
Electronic ISSN: 1756-9966
DOI
https://doi.org/10.1186/s13046-021-02173-5

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