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A Novel Immunocompetent Mouse Model of Pancreatic Cancer with Robust Stroma: a Valuable Tool for Preclinical Evaluation of New Therapies

  • 2015 SSAT Plenary Presentation
  • Published:
Journal of Gastrointestinal Surgery Aims and scope

Abstract

A valid preclinical tumor model should recapitulate the tumor microenvironment. Immune and stromal components are absent in immunodeficient models of pancreatic cancer. While these components are present in genetically engineered models such as KrasG12D; Trp53R172H; Pdx-1Cre (KPC), immense variability in development of invasive disease makes them unsuitable for evaluation of novel therapies. We have generated a novel mouse model of pancreatic cancer by implanting tumor fragments from KPC mice into the pancreas of wild type mice. Three-millimeter tumor pieces from KPC mice were implanted into the pancreas of C57BL/6J mice. Four to eight weeks later, tumors were harvested, and stromal and immune components were evaluated. The efficacy of Minnelide, a novel compound which has been shown to be effective against pancreatic cancer in a number of preclinical murine models, was evaluated. In our model, consistent tumor growth and metastases were observed. Tumors demonstrated intense desmoplasia and leukocytic infiltration which was comparable to that in the genetically engineered KPC model and significantly more than that observed in KPC tumor-derived cell line implantation model. Minnelide treatment resulted in a significant decrease in the tumor weight and volume. This novel model demonstrates a consistent growth rate and tumor-associated mortality and recapitulates the tumor microenvironment. This convenient model is a valuable tool to evaluate novel therapies.

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Acknowledgments

This study was funded by NIH grants R01-CA170946 and CA124723 (to AKS); NIH grant R01-CA184274 (to SB); Katherine and Robert Goodale foundation support (to AKS) and Minneamrita Therapeutics LLC (to AKS).

Author information

Authors and Affiliations

  1. Division of Basic and Translational Research, Department of Surgery, University of Minnesota, Minneapolis, MN, 55455, USA

    Kaustav Majumder, Nivedita Arora, Shrey Modi, Rohit Chugh, Alice Nomura, Bhuwan Giri, Rajinder Dawra, Sundaram Ramakrishnan, Sulagna Banerjee, Ashok Saluja & Vikas Dudeja

Authors
  1. Kaustav Majumder
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  2. Nivedita Arora
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  3. Shrey Modi
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  4. Rohit Chugh
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  5. Alice Nomura
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  6. Bhuwan Giri
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  7. Rajinder Dawra
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  8. Sundaram Ramakrishnan
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  9. Sulagna Banerjee
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  10. Ashok Saluja
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  11. Vikas Dudeja
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Corresponding author

Correspondence to Vikas Dudeja.

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Conflict of Interest

R Chugh has ownership interest in a patent of Minnelide. AK Saluja has ownership interest (including patents) in Minneamrita therapeutics and is a consultant/advisory board member for Minneamrita Therapeutics. The remaining authors declare no conflict of interest.

Grant Support

NIH grants R01-CA170946 and CA124723 (to AKS); NIH grant R01-CA184274 (to SB)

Ethical Approval

All procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of University of Minnesota.

Additional information

Author names in bold designate shared first co-first authorship.

Primary Discussant

O. Joe Hines, M.D. (Los Angeles, CA)

Congratulations to you Mr. Majumder and your colleagues from Minnesota. Your group reports an immunocompetent model for pancreatic cancer utilizing cells and tissue harvested from mice bearing mutant kRas and p53. I agree that your analysis of the model suggests that it recapitulates human pancreatic cancers with desmoplasia, an inflammatory infiltrate, and a typical pattern of metastases.

My questions are related to two broad areas—the method of implantation and implications of varying degrees of fibrosis based on this method.

1. OJH: Is it possible that single cell suspension injection resulted in a false incidence of metastatic lesions? We have found that after injection single cells can escape into the peritoneum implanting on various organs resulting in what appears to be a true metastasis but in fact is an aberration of this technique. What was the incidence of metastasis between the two techniques and if there was a difference do you think this issue influenced these findings?

2. OJH: You have made a very interesting observation that single cell suspension injection resulted in limited fibrosis while the tumor fragment model engendered significant desmoplasia. New evidence suggests that the process of desmoplasia during the early phases of pancreatic carcinogenesis may have a protective/beneficial impact. Desmoplasia may act as a scar slowing the conversion to frank cancer and in the case of cancer help to contain the cancer cells locally. The depletion of carcinoma-associated fibroblasts and fibrosis has been shown in induce immunosuppression and accelerate pancreatic cancer progression. Is it possible in your experiments that when injecting pure cancer cells this process is circumvented but when tumor fragments were used in your model, stromal elements that have participated in the process of carcinogenesis were introduced and may responsible for your findings of fibrosis that are similar to the KPC model? Was there a difference in disease burden and survival between these two groups? What do you hypothesize is the role of fibrosis in pancreatic cancer biology?

Closing Discussant

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1. KM: Thank you for your comments and questions, Dr. Hines. We have compared three immunocompetent models: the Syngeneic Tumor Implantation Model (STIM), genetically engineered KPC model and the previously used KPC tumor derived cell line implantation model. We would like to emphasize that the model where the single cell suspension generated from KPC tumors is injected has been reported before and we have used this model only as a comparison to the model developed by us. I agree with your comment about the single cell suspension injections contributing to a false incidence of metastatic lesions and we believe that this is one of the major limitations of this model. We found that although the mice injected with cell suspensions have peritoneal metastases and a shorter survival, they do not generally have liver or lung metastases as opposed to both the STIM model proposed by us and genetically engineered KPC models. We observed peritoneal metastases in 90 % of mice injected with single suspensions and in 80 % of STIM mice. However we observed liver metastases in 60 % and lung metastases in 20 % of our STIM mice as opposed to 0 % of single cell suspension injected mice. We therefore agree with you and have reason to believe that although the single cell suspension model has peritoneal metastases, they might be a result of direct cell spillage into the peritoneum since systemic metastases to lung and liver are not observed at the time of death.

2. KM: That is a great question. We do believe that since single cell suspensions only contain the epithelial component of pancreatic cancer, as opposed to the tumor fragment which contains both the epithelial and stromal components, the tumors that develop in the mice injected with single cell suspensions lack adequate desmoplastic reaction and therefore are more responsive to chemotherapeutic agents in preclinical studies (Olive et al., 2009). Considering the undeniable role of stroma in mediating tumor progression and chemoresistance, a valid tumor model should have stromal elements. In mice injected with single cell suspensions, mice became moribund by 4–6 weeks and 90 % of had peritoneal metastases when euthanized. Most of our STIM mice died within 3 months post-implantation, 80 % had peritoneal metastases, 60 % had liver metastases and 20 % had lung metastases. However we believe that our model recapitulates the disease progression of pancreatic cancer in a predictable manner and retains the metastatic properties of pancreatic cancer as evident by the presence of not only peritoneal metastases but also liver and lung metastases. Based on previous studies as well as our own experience, we have observed that once a tumor has been established, stroma is involved in both tumor progression and in mediating chemoresistance. We, like others before us, have observed that drugs that target both the epithelial and stromal components are more effective in decreasing tumor growth and metastasis. Since our model does not pass through the pre-invasive stages of pancreatic development, we cannot comment on the role of stroma in the tumorigenesis, especially in pre-invasive lesions such as PanINs.

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Majumder, K., Arora, N., Modi, S. et al. A Novel Immunocompetent Mouse Model of Pancreatic Cancer with Robust Stroma: a Valuable Tool for Preclinical Evaluation of New Therapies. J Gastrointest Surg 20, 53–65 (2016). https://doi.org/10.1007/s11605-015-2985-y

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