MRI for noninvasive characterization of pancreatic tumor in three mouse models
Navid Farr1, Yak-Nam Wang2, Samantha D’Andrea3, Kayla Gravelle3, Joo Ha Hwang1,3, and Donghoon Lee4

1Department of Bioengineering, University of Washington, Seattle, WA, United States, 2Applied Physics Laboratory, University of Washington, Seattle, WA, United States, 3Department of Medicine, University of Washington, Seattle, WA, United States, 4Department of Radiology, University of Washington, Seattle, WA, United States

Synopsis

The lethal nature of the pancreatic cancer is apparent in the 5-year survival rate of less than 5%. Choosing the right animal model is of utmost importance in oncology drug development research. There is a dire necessity for noninvasive techniques to monitor and characterize the tumors in different models to properly design drug delivery studies. We performed multi-parametric MRI at a high resolution to noninvasively identify the tumor and assess tumor volume characteristics. We used T1 and T2 relaxation, diffusion, and magnetization transfer effects along with 3 dimensional volume measurements. MR measurements were then compared with quantifiable histopathological results.

Target audience

Researchers working on new therapies for pancreatic cancer and groups working on preclinical studies with interest in noninvasive monitoring technique will benefit from this work.

Purpose

Pancreatic ductal adenocarcinoma (PDA) is among the most lethal human cancers. In addition, PDA is among the few cancers in which death rates have risen in the 21st century (1). Many conventional chemotherapeutic regimens have failed the standard of care, as only a minority of patients have had survival prolonged by several weeks (2,3). Oncology drug development relies heavily on mouse models bearing tumors for efficacy testing of delivery and efficacy of therapeutic agents. Therefore, choosing the right mouse model with characteristics similar to human PDA is essential. Also, noninvasive techniques are necessary to monitor and characterize the tumors in different models for properly designed drug delivery studies. Characterizing the tumor tissue can be used to predict the response to chemotherapy (4). The purpose of this study was to characterize the different properties of PDA tumors in three different mouse models. Alongside T2-weighted MRI, which was used to accurately identify and measure the size of the tumor, multi-parametric MR techniques were utilized to identify the tumor characteristics.

Methods

In addition to a genetically engineered KPC mouse model, which spontaneously develops PDA (5), we used orthotopic and subcutaneous xenograph mouse models. We conducted multi-parametric MRI at 14T (Paravision 5.1 software, Bruker Corp, Billerica, MA) to identify the tumor and surrounding organs and measured the stroma level and diffusion of each tumor model. MRI measurements include quantitative T2, ADC, magnetization transfer suppression ratio and three-dimensional (3D) volume measurements. Noninvasive MRI was performed for 8 animals of each tumor model. T2 relaxation, ADC values and tumor volumes were longitudinally recorded. The magnetization transfer ratio (MTR) was measured by utilizing a gradient echo sequence with an offset frequency of 7000 Hz. The MTR was compared to quantified fibrotic tissue measured from Masson’s trichrome staining of tumor samples.

Results and Discussion

T2 relaxation times of three tumor types were very similar but there was a statistically significant difference between the tumors and surrounding organs. Thus it will be helpful in detecting pathological tissues due to their elongated T2 relaxation times (Fig. 1). 3D reconstruction of each tumor is made possible by high resolution two-dimensional imaging, which generates thin multislice images that can then be used to calculate the volume of the tumor. ADC values of the tumor tissue in KPC tumor model were significantly lower than the other two types of orthotopic and subcutaneous models. This result might be associated with increased cellularity of the tumor in comparison to the other tumor models. Histology data revealed that the KPC model consistently demonstrated significantly greater levels of fibrosis and associated collagen deposition as shown in Fig. 2. The comparison of MTR and histologyc data indicates there is a correlation (r = 0.87) between MT ratio and the amount of stroma in each tissue (see Fig. 3). The data confirms that the MTR can be used as a noninvasive biomarker of stroma measurement.

Conclusions

Multi-parametric MRI utilizing T2, ADC, MTR and 3D volume measurements have feasibility in the identification and monitoring of the disease progression. Moreover, quantitative MR techniques are reliable noninvasive biomarkers of tumor characteristics.

Acknowledgements

This work was supported by NIH grants R01 CA154451 and R01 CA188654.

References

1. Siegel, R., Ma, J., Zou, Z. & Jemal, A. Cancer Statistics , 2014. 64, 9–29.

2. Burris 3rd, H. A. et al. Improvements in survival and clinical benefit with gemcitabine as first- line therapy for patients with advanced pancreas cancer: a randomized trial. J Clin Oncol. 1997;15, 2403–2413.

3. Tempero, M. et al. Randomized phase II comparison of dose-intense gemcitabine: thirty-minute infusion and fixed dose rate infusion in patients with pancreatic adenocarcinoma. J. Clin. Oncol. 2003; 21, 3402–8.

4. Niwa, T. et al. Advanced pancreatic cancer: the use of the apparent diffusion coefficient to predict response to chemotherapy. Br. J. Radiol. 2009;82, 28–34.

5. Hingorani, S. R. et al. Trp53 and Kras cooperate to promote chromosomal instability and widely metastatic pancreatic ductal adenocarcinoma in mice. Cancer Cell. 2005; 7, 469–483.

Figures

Figure 1. T2 relaxation time (ms) of the tumor overlaid onto a T2 weighted image. T2-map calculated semi-automatically from a series of T2 acquisitions.

Figure 2. Masson-trichrome-staining histology sections from tumors grown in each mouse model. (Left) KPC, (Middle) Orthotopic and (Right) Subcutaneous mice. Fibrotic stroma is depicted as blue-stained bands of collagen. Each image shows the overall distribution of fibrotic tissue within these central tumor slices.

Figure 3. MTR measurements (top) and histologic fibrotic tissue area measurements (bottom) for each of the three mouse models. There is a correlation between MT ratio and the amount of stroma in the tumor (r = 0.87).



Proc. Intl. Soc. Mag. Reson. Med. 24 (2016)
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