This article describes the utilization of high-resolution ultrasound in genetically engineered pancreatic cancer mice. The primary aim is to provide a detailed instruction for detection and evaluation of endogenous pancreatic tumors.
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Method Article
Utilizing High Resolution Ultrasound to Monitor Tumor Onset and Growth in Genetically Engineered Pancreatic Cancer Models
In This Article
Summary
Abstract
The LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre (KPC) mouse model represents an established and frequently used transgenic model to evaluate novel therapies in pancreatic cancer. Tumor onset is variable in the KPC model between 8 weeks and several months. Therefore, non-invasive imaging tools are required to screen for tumor onset and monitor for response to treatment. To address this issue, different approaches have emerged over the last years. High resolution ultrasound has major advantages such as non-invasiveness, fast session times and a high image resolution without radiation exposure. However, ultrasound in mice is not trivial and sufficient anatomical knowledge and practical skills are required to successfully perform high resolution ultrasound in preclinical pancreatic cancer models. With the following article, a detailed hands-on guide for abdominal ultrasound in murine models with a particular focus on endogenous pancreatic cancer models is presented. Furthermore, a summary of common mistakes and how to avoid them is provided.
Introduction
Genetically engineered mouse models have gained an increasing importance in cancer research due to their ability to closely recapitulate the complex nature of human carcinogenesis1,2,3. One of the most frequently used models to study pancreatic cancer development, progression and therapeutic response is characterized by an activating mutation in the Kras oncogene combined with an inactivation of the tumor suppressor p534. This LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre (KPC) mouse model mimics the step-wise progression f....
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Protocol
This protocol is in accordance with the animal care guidelines at the University Medical Centre Goettingen, Germany (33.9-42502-04-15/2056). Depending on specific requirements of individual animal review boards, some of the protocol steps could be modified accordingly.
1. Abdominal Palpation of KPC Mice
- To avoid unnecessary ultrasound scans, palpate the mouse abdomen to identify mice which might possibly bear an intraabdominal lesion and should subsequently undergo abdominal ultrasound.
- Perform abdominal palpation weekly starting from 8 weeks of age in the KPC mouse model. High resolution ultrasound can also be appli....
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Results
Ultrasound imaging is a versatile and non-invasive technique that is used to address several issues in murine models of human diseases. Compared to all other imaging approaches major advantages are high-throughput, cost efficiency, short acquisition time and real-time imaging. However, this tool needs expertise to generate accurate, high quality images. Particularly in the case of unwanted artefacts at least some experience with ultrasound imaging in general is very helpful. In relation t.......
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Discussion
With this protocol, a detailed description for quantifying pancreatic tumors using high-resolution abdominal ultrasound imaging in genetically engineered mouse models is provided. Recently, Sastra et al. published a detailed description how to quantify pancreatic tumors in mouse models, but no visualized instructions about the preparation and handling as prerequisite for all further steps were shown11. The overall goal of this manuscript is to provide a comprehensive visual guide for high.......
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Disclosures
The authors have nothing to disclose.
Acknowledgements
This research was supported by the Deutsche Krebshilfe (Max Eder Group to AN: 110972), a DGVS doctoral thesis scholarship (to SMB), and an Else-Kröner-Fresenius-Foundation scholarship (to RGG) at the University Medical Center Goettingen. We thank Jutta Blumberg and Ulrike Wegner for expert technical assistance. We also thank all animal technicians at the animal facility of the University Medical Center Goettingen for mouse keeping. All experiments were performed according to German animal welfare regulations.
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Visual Sonics Vevo2100 High Resolution Ultrasound System, including imaging stage and anesthesia line | FUJIFILM VisualSonics Inc, Canada | VS-11945 | |
| Vevo 2100 MicroScan Transducer MS-550-D (22-55MHz) | FUJIFILM VisualSonics Inc, Canada | VS-11874 | |
| Vevo Anesthesia System (anesthesia induction chamber with fresh and waste gas inlet) | FUJIFILM VisualSonics Inc, Canada | SA-12055 | |
| Vevo Imaging Station (working stage with nose cone for anesthesia supply) | FUJIFILM VisualSonics Inc, Canada | SA-11982 | |
| electronic pet clippers | Panasonic Marketing Europe, Germany | 5025232484324 | Panasonic ER-PA10-s |
| Labotect Hot plate | Labor tech Göttingen, Germany | 13854 | |
| eye cream (ophthalmic ointment) | Schülke&Mayr, Germany | 9080249 | |
| veterinary isoflurane | Abbvie, Germany | 4831867 | |
| depilatory cream | RB healthcare UK, United Kingdom | 8218535 | |
| 70% ethanol (v/v) in distilled water | TH. Geyer, Germany | 22941000 | |
| ultrasound gel | Asmuth, Germany | 13477 | |
| tissue wipes | Kimberly-Clark Germany, Germany | 7558 | |
| cotton tips | Meditrade, Germany | 75481116 | |
| glass bowl for ultrasound gel | ARC France, France | H1149 | |
| water bowl | W & P Trading Co., USA | B00K2P6PLQ | |
| gauze sponges | Fuhrmann, Germany | 960504 |
References
- Kersten, K., de Visser, K. E., van Miltenburg, M. H., Jonkers, J. Genetically engineered mouse models in oncology research and cancer medicine. EMBO Molecular Medicine. 9 (2), 137-153 (2017).
- Olive, K. P., Politi, K. Translational therapeutics in genetically engin....
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