Source: UNIVERSITY OF NEVADA submitted to NRP
HEALTH BENEFITS OF FREEZE-DRIED WATERCRESS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1030520
Grant No.
2023-67018-39757
Cumulative Award Amt.
$300,000.00
Proposal No.
2022-09496
Multistate No.
(N/A)
Project Start Date
Apr 1, 2023
Project End Date
Mar 31, 2026
Grant Year
2023
Program Code
[A1343]- Food and Human Health
Recipient Organization
UNIVERSITY OF NEVADA
(N/A)
RENO,NV 89557
Performing Department
(N/A)
Non Technical Summary
Pancreatic cancer is often diagnosed at an advanced stage and it is highly resistant to conventional chemotherapy and radiation therapy which makes it difficult to treat. The progression from normal pancreatic epithelium to pancreatic cancer is associated with the accumulation of several genetic mutations and gradually evolves via precursor lesions, called pancreatic intraepithelial neoplasia (PanIN) lesions. Epidemiological studies have suggested that consumption of cruciferous vegetables is associated with a lower risk of developing cancers, including pancreatic cancer.The overarching goal of this project is to characterize the efficacy of oral administration of the cruciferous vegetable watercress (Nasturtium officinale) on preventing the development and progression of early lesions of pancreatic intraepithelial neoplasia to more advanced lesions and pancreatic cancer, and on modulating the gut microbiome. The specific aims of this project are (1) to determine the efficacy of freeze-dried watercress on preventing the development and progression of precursor PanIN lesions to invasive pancreatic adenocarcinoma, by using a transgenic animal model which resembles histopathological features of the human disease, and (2) to characterize the effect of freeze-dried watercress consumption on the gut microbiome using metagenomics sequencing.This study will provide information regarding biomarkers of response to freeze-dried watercress in a pancreatic cancer model. The long-term goal is to develop a strategy to prevent the progression of PanIN lesions to pancreatic cancer using a whole-food approach.
Animal Health Component
50%
Research Effort Categories
Basic
50%
Applied
50%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
70214991010100%
Goals / Objectives
The overarching goal of the project is to characterize the efficacy of watercress (Nasturtium officinale) on preventing the development and progression of early lesions of pancreatic intraepithelial neoplasia to more advanced lesions and pancreatic cancer, and on modulating the gut microbiome.The objectives of this project are (1) 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 which resembles histopathological features of the human disease, and (2) to characterize the effect of freeze-dried watercress consumption on the gut microbiome by shotgun metagenomics sequencing.
Project Methods
Specific Aim 1. Determine the efficacy of oral administration of freeze-dried watercress to prevent the development and progression of pancreatic intraepithelial neoplasia (PanIN) lesions to pancreatic cancer using the LSL-KrasG12D/+;Pdx1Cre/+ transgenic mouse model.The methods used in specific aim 1 will involve the transgenic animal model LSL-KrasG12D/+;Pdx1Cre/+ which resembles histopathological features of human pancreatic cancer. 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 makes this model suitable for prevention and interception studies because it provides a long period of intervention during the early, preinvasive states. This specific aim will evaluate whether oral administration of freeze-dried watercress prevents the onset and/or progression of PanIN lesions to invasive pancreatic adenocarcinoma.The preventive effect of the watercress intervention compared with vehicle (control) will be assessed based on the following primary and secondary endpoints. Primary endpoints: (1) incidence and histopathological grading of PanIN lesions; and (2) incidence of pancreatic cancer; and Secondary endpoints: (3) Analysis of proliferation, apoptosis, and cell cycle markers in pancreatic tissue. Pancreas weights and incidence of metastasis will be also recorded and compared between watercress-treated group and control group.Specific Aim 2. Determine the impact of oral administration of freeze-dried watercress on the gut microbiome using shotgun metagenomic sequencing analysis.The methods used in specific aim 2 will involve shotgun metagenomic sequencing analysis of the gut microbiome. The shotgun metagenomic sequencing is used to determine the effect of watercress administration on the gut microbiome taxonomic composition, and to characterize the relative abundance of taxa as compared to vehicle-treated control.Overall, this study will provide information necessary in developing biomarkers of response to freeze-dried watercress and it will characterize the impact of freeze-dried watercress on the gut microbiome.

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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