Name: ultrasound imaging-guided intracardiac injection to develop a mouse model of breast cancer brain metastases followed by longitudinal mri
Uploaded: 2014-03-06T13:45:00
Description: Watch this Scientific Journal Video about Ultrasound Imaging-guided Intracardiac Injection to Develop a Mouse Model of Breast Cancer Brain Metastases Followed by Longitudinal MRI at JoVE.com

We are grateful to Drs. Diane Palmieri and Patricia Steeg of NCI for providing us MDA-MB231Br cells. We thank Dr. Ralph Mason, Mr. Jason Reneau and Ms. Ramona Lopes for technical and collegial support. This work was supported in part by the DOD IDEA Awards W81XWH-08-1-0583 and W81XWH-12-1-0317. MRI experiments were performed in the Advanced Imaging Research Center, an NIH BTRP # P41-RR02584 facility, and ultrasound-guided intracardiac injection was performed with VisualSonics Vevo 770 under 1S10RR02564801.

All animal procedures were approved by the Institutional Animal Care and Use Committee of University of Texas Southwestern Medical Center.
1. Preparation of the MDA-MB231/Br-GFP Cells
<ol>
	<li>Retrieve and culture the MDA-MB231/Br-GFP cells (kindly provided by Drs. Palmieri and Steeg, NCI) in DMEM medium containing 10% FBS, 1% glutamine and 1% penicillin/streptomycin.</li>
	<li>Observe the condition of the cells and color of medium, and replace medium every 2-3 days.</li>
	<li>Trypsinize an...

<ol>
	<li>Schouten, L. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidence+of+brain+metastases+in+a+cohort+of+patients+with+carcinoma+of+the+breast,+colon,+kidney,+and+lung+and+melanoma.">Incidence of brain metastases in a cohort of patients with carcinoma of the breast, colon, kidney, and lung and melanoma.</a> Cancer. 94, 2698-2705 (2002).</li><li>Lin, N. U., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CNS+metastases+in+breast+cancer.">CNS metastases in breast cancer.</a> J. Clin. Oncol. 22, 3608-3617 (2004).</li><li>Eichler, A. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biology+of+brain+metastases-translation+to+new+therapies.">The biology of brain metastases-translation to new therapies.</a> Nat. Rev. Clin. Oncol. , (2011).</li><li>Lockman, P. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterogeneous+blood-tumor+barrier+permeability+determines+drug+efficacy+in+experimental+brain+metastases+of+breast+cancer.">Heterogeneous blood-tumor barrier permeability determines drug efficacy in experimental brain metastases of breast cancer.</a> Clin. Cancer Res. 16, 5664-5678 (2010).</li><li>Yoneda, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+bone-seeking+clone+exhibits+different+biological+properties+from+the+MDA-MB-231+parental+human+breast+cancer+cells+and+a+brain-seeking+clone+in+vivo+and+in.">A bone-seeking clone exhibits different biological properties from the MDA-MB-231 parental human breast cancer cells and a brain-seeking clone in vivo and in.</a> 16, 1486-1495 (2001).</li><li>Palmieri, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Her-2+overexpression+increases+the+metastatic+outgrowth+of+breast+cancer+cells+in+the+brain.">Her-2 overexpression increases the metastatic outgrowth of breast cancer cells in the brain.</a> Cancer Res. 67, 4190-4198 (2007).</li><li>Percy, D. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+characterization+of+changing+blood-tumor+barrier+permeability+in+a+mouse+model+of+breast+cancer+metastasis:+a+complementary+magnetic+resonance+imaging+approach.">In vivo characterization of changing blood-tumor barrier permeability in a mouse model of breast cancer metastasis: a complementary magnetic resonance imaging approach.</a> Invest. Radiol. 46, 718-725 (2011).</li><li>Zhou, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longitudinal+MRI+evaluation+of+intracranial+development+and+vascular+characteristics+of+breast+cancer+brain+metastases+in+a+mouse+model.">Longitudinal MRI evaluation of intracranial development and vascular characteristics of breast cancer brain metastases in a mouse model.</a> PLoS One. 8, (2013).</li><li>Zhou, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+near-infrared+optical+imaging+of+2-deoxyglucose+uptake+by+intracranial+glioma+of+athymic+mice.">Dynamic near-infrared optical imaging of 2-deoxyglucose uptake by intracranial glioma of athymic mice.</a> PLoS One. 4, (2009).</li><li>Zhang, R. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+permeability+of+the+blood-brain+barrier+in+experimental+brain+metastases+produced+by+human+neoplasms+implanted+into+nude+mice.">Differential permeability of the blood-brain barrier in experimental brain metastases produced by human neoplasms implanted into nude mice.</a> Am. J. Pathol. 141, 1115-1124 (1992).</li><li>Mason, R. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+oximetry:+comparison+of+19F+MR+EPI+and+electrodes.">Tumor oximetry: comparison of 19F MR EPI and electrodes.</a> Adv. Exp. Med. Biol. 530, 19-27 (2003).</li><li>Gril, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pazopanib+reveals+a+role+for+tumor+cell+B-Raf+in+the+prevention+of+HER2++breast+cancer+brain+metastasis.">Pazopanib reveals a role for tumor cell B-Raf in the prevention of HER2+ breast cancer brain metastasis.</a> Clin. Cancer Res. 17, 142-153 (2010).</li></ol>

In the present study, we have demonstrated that ultrasound imaging-guided left ventricular injection ensures the accuracy so that every animal in this study developed brain metastases and no animal death was observed.&#160;The beauty of image-guided injection is that the path of needle penetration of skin and finally into the left ventricle can be monitored and adjusted under image, which is distinct from the manual procedure requiring strict anatomic landmarks to follow.
Current understanding...

Brain metastasis is the most common intracranial malignancy in adults. The prognosis is extremely poor, with a median survival of 4-6 months even with aggressive treatment. Breast cancer is one of the three major primary cancers with a high morbidity of brain metastasis1-3. Several brain-tropic breast cancer lines are capable of developing brain metastases after intracardiac or intracarotid injection4. Direct injection of tumor cells into the left ventricle can bypass the lung capillary bed and thus increase the incidence of forming brain metastases while minimizing visceral metastases.&#160;The MDA-MB231Br line is one of the most widely used human breast cancer lines to develop brain metastasis in rodent models5,6.
Like many other studies4,7, we have performed a manual procedure of intracardiac injection in our previous studies. However, only 50% successful rate was obtained with the manual injection and a fraction of mice died from the repeated invasive procedures if prior trials failed. Here, we are introducing the use of an imaging-guided procedure to secure the injection of brain-seeking breast cancer cells into the left ventricle of athymic mice. Longitudinal high resolution MRI is applied to follow intracranial development of brain metastases.

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			<td height="21" style="height:21px;width:413px;">DMEM</td>
			<td style="width:136px;">HyClone</td>
			<td style="width:87px;">SH30022.01</td>
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			<td height="21" style="height:21px;width:413px;">VisualSonics Vevo 770 High-Resolution Imaging System </td>
			<td>Visual Sonics Inc.</td>
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			<td height="21" style="height:21px;">9.4T horizontal bore magnet with a Varian INOVA Unity system </td>
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ultrasound imaging-guided intracardiac injection to develop a mouse model of breast cancer brain metastases followed by longitudinal mri

Breast cancer brain metastasis, occurring in 30% of breast cancer patients at stage IV, is associated with high mortality. The median survival is only 6 months. It is critical to have suitable animal models to mimic the hemodynamic spread of the metastatic cells in the clinical scenario. Here, we are introducing the use of small animal ultrasound imaging to guide an accurate injection of brain tropical breast cancer cells into the left ventricle of athymic nude mice. Longitudinal MRI is used to assessing intracranial initiation and growth of brain metastases. Ultrasound-guided intracardiac injection ensures not only an accurate injection and hereby a higher successful rate but also significantly decreased mortality rate, as compared to our previous manual procedure. In vivo high resolution MRI allows the visualization of hyperintense multifocal lesions, as small as 310 &#181;m in diameter on T2-weighted images at 3 weeks post injection. Follow-up MRI reveals intracranial tumor growth and increased number of metastases that distribute throughout the whole brain.

With the high spatial resolution of MRI (78 &#181;m in plane resolution), hyperintense lesions can be identified as small as 310 &#181;m in diameter (Figure&#160;2). Since the metastases in this study are very small and development of necrosis and edema is minimal, the hyperintense lesion on T2-weighted images truly represented the tumor mass.
Longitudinal MRI studies allow in vivo noninvasive evaluation of tumor growth. As shown in Figure 3</strong...

Watch this Scientific Journal Video about Ultrasound Imaging-guided Intracardiac Injection to Develop a Mouse Model of Breast Cancer Brain Metastases Followed by Longitudinal MRI at JoVE.com

Ultrasound Imaging-guided Intracardiac Injection to Develop a Mouse Model of Breast Cancer Brain Metastases Followed by Longitudinal MRI

A breast cancer brain metastasis mouse model is established with ultrasound imaging-guided intracardiac injection of MDA-MB231/Br-GFP cells. Development of multifocal intracranial metastases has been monitored longitudinally using high-resolution 9.4 T MRI.

Breast cancer brain metastasis, occurring in 30% of breast cancer patients at stage IV, is associated with high mortality. The median survival is only 6 ...

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