A novel animal model of human breast cancer metastasis to the spine: a pilot study using intracardiac injection and luciferase-expressing cells

Presented at the 2012 Spine Section Meeting 

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Object

Metastatic spine disease is prevalent in cancer victims; 10%–30% of the 1.2 million new patients diagnosed with cancer in the US exhibit spinal metastases. Unfortunately, treatments are limited for these patients, as disseminated disease is often refractory to chemotherapy and is difficult to treat with surgical intervention alone. New animal models that accurately recapitulate the human disease process are needed to study the behavior of metastases in real time.

Methods

In this study the authors report on a cell line that reliably generates bony metastases following intracardiac injection and can be tracked in real time using optical bioluminescence imaging. This line, RBC3, was derived from a metastatic breast adenocarcinoma lesion arising in the osseous spine of a rat following intracardiac injection of MDA-231 human breast cancer cells.

Results

Upon culture and reinjection of RBC3, a statistically significantly increased systemic burden of metastatic tumor was noted. The resultant spine lesions were osteolytic, as demonstrated by small animal CT scanning.

Conclusions

This cell line generates spinal metastases that can be tracked in real time and may serve as a useful tool in the study of metastatic disease in the spine.

Abbreviations used in this paper:BLI = bioluminescence imaging; FBS = fetal bovine serum; SARRP = small animal radiation research platform; VB = vertebral body.

Article Information

Address correspondence to: Daniel Sciubba, M.D., 600 North Wolfe Street, Meyer 5-185, Baltimore, Maryland 21287. email: dsciubb1@jhmi.edu.

Please include this information when citing this paper: published online December 21, 2012; DOI: 10.3171/2012.11.SPINE12325.

© AANS, except where prohibited by US copyright law.

Headings

Figures

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    Bioluminescence of MDA-231 cells following intracardiac injection. Rats are shown on Days 10 (A), 34 (B), and 46 (C) after intracardiac injection of MDA-231 cells. A: Bioluminescence positivity is diffuse on Day 10. B: By Day 34, 3 animals demonstrate focal bioluminescence positivity in the spine. C: The surviving animals are shown on Day 46, with bioluminescence in the forepaw, femur, and hindpaw.

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    Small animal CT scanning and bioluminescent signal of osteolytic metastasis. A: Bioluminescence signal in lumbar spine of 1 rat following intracardiac injection of MDA-231 cells. The inset highlights full body metastasis. B: Small animal CT scanning confirms osteolysis of the posterior elements of lumbar spine (arrow) with decrease in calcium in the location of bioluminescent signal.

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    Bioluminescence of MDA-231 cells after intracardiac injection. A: Animals are shown on Day 34 after intracardiac injection. Metastases are evident in the spine in all 3 animals. Additional metastases are noted in the anatomical locations of the scapula, lung, femur, forepaw, tail, hindpaw, and brain. B and C: Cells were recovered from the spinal lesions in Rat 3 (A, right). These cells were luciferase (B) and GFP (C) positive, confirming their source as the MDA-231 cell line.

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    Bioluminescence of RBC3 cells following intracardiac injection. Rats are shown on Days 11 (A), 34 (B), and 43 (C) after intracardiac injection of RBC3 cells. A: Bioluminescence positivity is diffuse on Day 11. B: By Day 34, 6 animals demonstrate focal bioluminescence positivity in the spine. C: On Day 43, the remaining animals demonstrate marked progression of metastatic disease to bilateral femurs, multiple foci in the lumbar spine, scapula, and fore- and hindpaws. One animal did not demonstrate bioluminescence at any time during the study.

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    Small animal CT scanning and bioluminescent signal of osteolytic metastasis. A: Bioluminescence signal in lumbar spine of rat following intracardiac injection of RBC3 cells. B: Computed tomography scan confirming osteolysis of the posterior elements of lumbar spine (arrow) in the location of the bioluminescent signal.

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    Kaplan-Meier survival curve. Survival times to paralysis for MDA-231 and RBC3 intracardiac-injected rats are shown. The difference between the curves was not statistically significant (p = 0.277).

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    Quantification of the total tumor burden. Tumor burden was measured in radiance on Day 34 for all animals. The average tumor burden is shown. Error bars indicate standard deviation. There was a statistically significant difference in tumor burden between the groups (p = 0.013, Student t-test).

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    Photomicrographs of metastatic lesions. Metastatic lesions were excised en bloc and stained with H & E for analysis. A–C: Aggressive invasion of metastatic adenocarcinoma into the immature trabecular bone can be seen. D: Complete ablation of the bony trabeculae with invasion of the musculature. Specimens from an L-5 lesion on Day 34 (A), L-1 on Day 43 (B), L-2 on Day 43 (C), and S-1 on Day 53 (D) are shown. All lesions were excised from RBC3 intracardiac-injected animals.

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