Name: high-resolution ultrasonography for the analysis of orthotopic atc tumors in a genetically engineered mouse model
Uploaded: 2022-10-11T15:00:00
Description: Watch this Scientific Journal Video about High-Resolution Ultrasonography for the Analysis of Orthotopic ATC Tumors in a Genetically Engineered Mouse Model at JoVE.com

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The present study was performed with approval from the Animal Care and Use Committee of Sichuan University. TPO-cre/ERT2; BrafCA/wt; Trp53&#916;ex2-10/&#916;ex2-10&#160;transgenic mice10 were used in this study (see Table of Materials). The protocol steps can be modified for different animal species&#160;if necessary. Twelve mice, including six females and six males, with a mean age of 93 days, were used here.
1. Exp...

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This protocol uses high-resolution ultrasonography to analyze orthotopic ATC tumors in a genetically engineered mouse model. The transgenic model, with a genotype of TPO-cre/ERT2; BrafCA/wt; Trp53&#916;ex2-10/&#916;ex2-10, was developed in our laboratory. The animals overexpress BrafV600E and lack Trp53; injecting the animals intraperitoneally with tamoxifen leads to tumor growth after approximately 1 month10. The tumors grow rapidly and reach a m...

Although anaplastic thyroid carcinoma (ATC) accounts for fewer than 2% of thyroid cancers, it causes more than 50% of thyroid cancer-related deaths annually. The median survival time after diagnosis with ATC is only about 6 months, and no treatments are available that significantly improve survival1,2.
The rarity of ATC has hampered the research studying how the disease begins and aggressively progresses. Genetically engineered mouse models that mimic the disease have recently become available, which provide insights into the disease and its responses to possible treatments3,4,5. Such studies require accurate tumor imaging for measurements and monitoring, which is typically performed using magnetic resonance imaging, computed tomography, or high-resolution ultrasonography6,7. Ultrasonography has been widely used in mouse organs. It has advantages over magnetic resonance imaging and computed tomography since it can be performed in real-time and does not expose the subject to radiation, and the necessary equipment is small enough to be portable8,9. However, studies on continuously monitoring ATC growth using ultrasound are rare; therefore, this work explores the utility of ultrasound in this context.
Here, a protocol for using high-resolution ultrasonography to accurately scan, monitor, and measure tumors in a mouse model of ATC is presented.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Adhesive tape</td>
			<td>Winner</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthesia system</td>
			<td>RWDlifescience</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Brafflox/wt mice</td>
			<td>Collaboration with Institute of Life Science, eBond Pharmaceutical Technology Ltd, Chengdu, China</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Chamber for anesthesia induction</td>
			<td>RWDlifescience</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton swabs</td>
			<td>Winner</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Depilatory cream</td>
			<td>Veet</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Electric heating blanket</td>
			<td>Petbee</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane vaporizer</td>
			<td>RWDlifescience</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Medical gloves</td>
			<td>Winner</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Paper towels</td>
			<td>Breeze</td>
			<td>B914JY</td>
			<td></td>
		</tr>
		<tr>
			<td>TPO-cre/ERT2 mice</td>
			<td>Collaboration with Institute of Life Science, eBond Pharmaceutical Technology Ltd, Chengdu, China</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Trp53flox/wt mice</td>
			<td>Collaboration with Institute of Life Science, eBond Pharmaceutical Technology Ltd, Chengdu, China</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound gel</td>
			<td>Keppler</td>
			<td>KL-250</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound machine</td>
			<td>VisualSonics</td>
			<td>Vevo 3100</td>
			<td></td>
		</tr>
	</tbody>
</table>

high-resolution ultrasonography for the analysis of orthotopic atc tumors in a genetically engineered mouse model

Anaplastic thyroid carcinoma (ATC) is associated with a poor prognosis and short median survival time, but no effective treatment improves the outcomes significantly. Genetically engineered murine models that mimic ATC&#39;s progression may help researchers to study treatments for this disease. Crossing three different genotypes of mice, a TPO-cre/ERT2; BrafCA/wt; Trp53&#916;ex2-10/&#916;ex2-10&#160;transgenic ATC model was developed. The ATC murine model was induced by an intraperitoneal injection of tamoxifen with overexpression of BrafV600E and deletion of Trp53, and the tumors were generated within about 1 month. High-resolution ultrasound was applied to investigate the tumor initiation and progression, and the dynamic growth curve was obtained by measuring the tumor sizes. Compared to magnetic resonance imaging (MRI) and computed tomography scanning, ultrasound has advantages in observing the ATC murine model, such as being noninvasive, portable, in real-time, and without radiation exposure. High-resolution ultrasound is suitable for dynamic and multiple measurements. However, ultrasonographic examination of the thyroid in mice requires relevant anatomical knowledge and experience. This article provides a detailed procedure for utilizing high-resolution ultrasound to scan tumors in the transgenic ATC model. Meanwhile, ultrasonic parameter adjustment, ultrasound scanning skills, anesthesia and recovery of the animals, and other elements that need attention during the process are listed.

The average right ATC size at the beginning of the study was 4.867 mm2, and the average left ATC size was 5.189 mm2. On the fourth measurement, the average right ATC size had grown to 11.844 mm2, while the tumor size of the left lobe had grown to 9.280 mm2. The total ATC size increased from 10.057 mm2 to 15.843 mm2. In the later stage of the study, the ATC grew rapidly. In terms of the mouse labeled &#34;P92&#34; (Table 1), the tumor size ...

Watch this Scientific Journal Video about High-Resolution Ultrasonography for the Analysis of Orthotopic ATC Tumors in a Genetically Engineered Mouse Model at JoVE.com

High-Resolution Ultrasonography for the Analysis of Orthotopic ATC Tumors in a Genetically Engineered Mouse Model

The present protocol describes high-frequency ultrasonography for visualizing the entire mouse thyroid gland and monitoring the growth of anaplastic thyroid carcinoma.

Anaplastic thyroid carcinoma (ATC) is associated with a poor prognosis and short median survival time, but no effective treatment improves the outcomes ...

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