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

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.

Zebrafish patient-derived xenografts accurately and quickly reproduce treatment outcomes in non–small cell lung cancer patients

“…However, there are few studies on lung cancer drug screening. Fan et al 21 explored the inhibitory effect of dosimertinib and gefitinib in a zebrafish brain metastasis model. Li et al 20 used the zPDX model to study the inhibitory effect of osimertinib on NSCLC with EGFR mutation and T790M resistance mutation.…”

“…We set up four to six concentration gradients for each drug and set the reference concentration based on our previous experience and published research. [19][20][21][22] Although there is no clear conversion formula, the maximum plasma concentration of medication for patients has also been used as a reference to set the reference concentration. 13,17 However, the final MTD concentration is still to be determined by setting the concentration gradient and conducting toxicity tests in zebrafish embryos.…”

RPTOR blockade suppresses brain metastases of NSCLC by interfering the ceramide metabolism via hijacking YY1 binding.

“…To observe whether abnormal RPTOR expression promotes BM in the host, the zebra sh BM model was established by overexpressing or knocking down RPTOR in zebra sh eggs and subsequently injecting lung cancer cells into the embryos, as previously described [32,33]. Brie y, zebra sh eggs at the unicellular stage of the Tg (coro1a:EGFP) line were recruited.…”

Zebrafish Xenograft Model for Studying Pancreatic Cancer-Instructed Innate Immune Microenvironment

“…It shows a high degree of physiological and genetic similarity to mammals and can be used to mimic the tumor microenvironment. Zebrafish xenotransplantation model is widely used to evaluate cancer progression, neo-angiogenesis, and drug response/resistance [ 23 , 24 , 25 ]. In zebrafish, the innate and adaptive immune system is highly conserved [ 26 ], and the cancer cells and innate immune cells can be fluorescently labelled and directly observed in the living animal due to optical clarity of zebrafish larvae.…”