Progress 04/01/24 to 03/31/25
Outputs
Target Audience:This seed project provides a proof-of-principle on the efficacy of freeze-dried watercress in preventing the development and progression of pancreatic intraepithelial neoplasia lesions, and therefore the outcomes of this project are relevant for individuals who present pancreatic intraepithelial neoplasia lesions and have a high risk of developing pancreatic cancer. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?Poster presentation at a national conference, NUTRITION 2025(P12-025-25): Stan, S.D. Freeze-dried watercress consumption modulates the gut microbiota in mice. Current Developments in Nutrition 9 (2025) 106391. What do you plan to do during the next reporting period to accomplish the goals?Finalize the research activities listed under objectives 1 and 2 of the project and submitmanuscripts for publication.
Impacts
What was accomplished under these goals?
We performed research activities listed under objectives 1 and 2 of the project. Objective 1 of the project is to determine the efficacy of freeze-dried watercress to prevent the development of precursor pancreatic intraepithelial neoplasia lesions to invasive pancreatic adenocarcinoma, by using the transgenic animal model LSL-KrasG12D/+;Pdx1Cre/+. In this model, mice develop pancreatic cancer within a physiologic microenvironment and present progressive stages of pancreatic cancer starting from early precursor intraepithelial hyperplasia lesions PanIN-1 to intermediate lesions PanIN-2, and then to advanced PanIN-3 lesions which can progress to pancreatic adenocarcinoma in older mice. The progressive development to high-grade PanIN lesions over several months makes this model suitable for prevention and interception studies because it provides the opportunityof intervention during the early, preinvasive stages. During this reporting period, it was accomplished (1) the expansion of the colony of transgenic mice, (2) the randomized assignment of the mice to either the watercress intervention group or the control group, and (3) the progression of treatment which iscurrently underway. In this model, advanced PanIN lesions and pancreatic adenocarcinoma occur in the control mice atapproximately 38-44 weeks of age. Objective 2 of the project is to determine the impact of oral administration of freeze-dried watercress on the gut microbiome using shotgun metagenomic sequencing analysis. Although the treatment study is currently underway, we performed an early, preliminary investigation of the gut microbiota using shotgun metagenomic sequencing analysis at two early time points, to set up the methodology protocol for objective 2 and to obtain an early characterization of the impact of oral administration of freeze-dried watercress on the gut microbiome. Differential abundance of taxa was analyzed using the One Codex microbiome analysis platform. The results were presented as a poster at the NUTRITION 2025 conference (P12-025-25).
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Progress 04/01/23 to 03/31/24
Outputs
Target Audience:This project will provide a proof-of-principle for the efficacy of freeze-dried watercress to prevent the development and progression of pancreatic intraepithelial neoplasia lesions, and therefore the outcomes of this project are relevant for individuals who present pancreatic intraepithelial neoplasia lesions and have a high risk of developing pancreatic cancer. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Trained one graduate student (Nutrition Science). How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?Performresearch activities under objectives 1 and 2 of the project as described under the experimental design for this project. 1. Continue the transgenic animal study as described in the experimental design for specific aim1. 2. Gut microbiome analysis by shotgun metagenomicsequencing analysis as described in the experimental design for specific aim2.
Impacts
What was accomplished under these goals?
We performed research activities under objective 1 of the project. Objective 1 of the project is to determine the efficacy of freeze-dried watercress to prevent the development of precursor pancreatic intraepithelial neoplasia lesions to invasive pancreatic adenocarcinoma, by using a transgenic animal model. Transgenic animal model LSL-KrasG12D/+;Pdx1Cre/+: The methodused to accomplish objective 1 involves the transgenic animal model LSL-KrasG12D/+;Pdx1Cre/+. In this model, mice develop pancreatic cancer within a physiologic microenvironment and present progressive stages of pancreatic cancer starting from early precursor intraepithelial hyperplasia lesions PanIN-1 to intermediate lesions PanIN-2, and then to advanced PanIN-3 lesions which can progress to pancreatic adenocarcinoma in older mice. The progressive development to high-grade PanIN lesions over several months makes this model suitable for prevention and interception studies because it provides a long period of intervention during the early, preinvasive states. In this model, pancreatic adenocarcinoma occurs by approximately 40-44 weeks of age. Breeding pairs of transgenic mice were purchased from the Jackson Laboratories and the transgenic animal study is underway. Preparation of freeze-dried watercress: Two commercially available samples of watercress, produced by two different growers, were purchased from local stores, freeze-dried, and then analyzed for the amount of the bioactive and cancer chemopreventive compound phenethyl isothiocyanate (PEITC).Freeze-dried watercress was grinded into a powder and then stored at -20°C. Scale-up production of freeze-dried watercress for the entire study is underway. Two sets of experimentswere completed for the freeze-dried watercress analysis: (1) Determination of PEITC amount in freeze-dried watercressby HPLC and GC-MS. Both HPLC and GC-MS analyses identified significant amounts of PEITC in the freeze-dried watercress samples. (2) Testing of freeze-dried watercress for mouse pathogens by PCR evaluation (Opti-XXpress/EDx Mouse Global Profile); thisanalysis wasneeded since the freeze-dried watercress is administered to mice. The PCRevaluation offreeze-dried watercressidentified no mouse pathogens. In addition, we developed a new and better method to administer freeze-dried watercress to mice by incorporating the freeze-dried watercressinto the diet. AIN-93M diet in powder form was mixed with freeze-dried watercress powder and the freeze-dried watercress-diet pellets were formulated as a replacement method for the daily oral gavage method of freeze-dried watercress which was initially proposed in this project. The PCRevaluation (Opti-XXpress/EDx Mouse Global Profile)of the freeze-dried watercress-diet pellets identified no mouse pathogens. One manuscript submitted: A review manuscript on the effects of the gut microbiome metabolites on cancer was submitted. Citation:Soto, J.P. and Stan, S.D. Effect of bacterial metabolites and probiotic strains on cancer. International Journal of Molecular Sciences (submitted).
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