Recipient Organization
TUFTS UNIVERSITY
28 SAWYER AVE
MEDFORD,MA 02155-5811
Performing Department
(N/A)
Non Technical Summary
As novel foods such as plant-based meats, cultivated meats, and other alternative proteins gain popularity for their potential to improve sustainability and reduce environmental impact, concerns remain about how these foods affect human health. Currently, food safety evaluations often rely on animal testing, which does not always accurately reflect how human bodies respond to new foods and present ethical and regulatory challenges. Moreover, the long-term health effects of novel foods, including their potential to trigger inflammation, allergies, or other chronic conditions, remain poorly understood. This project addresses the critical need for better, human-relevant tools to assess the safety and health impact of emerging foods before they reach the market. Ensuring the safety of our food supply is vital not only to individual health, but also to public trust, economic security, and the sustainable growth of the food industry.To solve this challenge, our team will build and use advanced laboratory models that mimic the human digestive system using human cells and tissues. We will simulate digestion and analyze how different novel foods interact with human gut tissues. These models will allow us to observe how cells respond, such as whether they show signs of stress, inflammation, or changes in function, without the need for animal testing. We will also study how long-term exposure to certain foods might change how our gut cells behave at a genetic level, using tools that measure changes in DNA and gene activity. Importantly, these tests will be designed using cells from multiple human donors to reflect a wide range of biological responses and increase relevance to real-world populations.By the end of this project, we aim to deliver an innovative, fully in vitro (lab-based) testing platform that can be used by scientists, regulators, and industry to evaluate the safety and biological effects of new food products. These tools will improve the reliability and efficiency of food safety research, reduce reliance on animal studies, and help manufacturers bring healthier, safer products to market. The broader societal benefits include greater consumer confidence, more ethical research practices, and stronger scientific foundations for future food innovation.
Animal Health Component
40%
Research Effort Categories
Basic
40%
Applied
40%
Developmental
20%
Goals / Objectives
Project Goal:The major goal of this project is to develop and apply an advanced, animal-free, in vitro human digestive and intestinal tissue model system to evaluate the safety, biological effects, and epigenetic impact of novel foods, including cultivated meat, plant-based proteins, and alternative dietary products. This system will serve as a standardized platform for comparative food safety assessment, enabling mechanistic insights and data generation to support regulatory and scientific evaluation of emerging food products.Objectives:Develop a physiologically relevant in vitro digestion pipeline using refined INFOGEST 2.0 methods incorporating human-derived enzyme activity, bile salt concentrations, and digestion kinetics tailored to whole foods. This pipeline will simulate digestion from oral intake to intestinal assimilation and support consistent feeding of the intestinal tissue model.Evaluate the biological impacts of digested novel foods on human intestinal tissue models. These impacts include epithelial integrity, mucus production, immune and allergenic responses, and microbiome changes. Responses will be measured at both acute and chronic timepoints to capture dynamic and persistent effects.Characterize epigenetic modifications in intestinal tissues following repeated exposure to novel food digests. This will involve profiling chromatin accessibility, DNA methylation, and histone modifications, and linking these changes to gene expression and functional outcomes observed in the intestinal model.
Project Methods
Aim 1: In Vitro Digestion Pipeline DevelopmentAn advanced digestion system will be constructed based on the INFOGEST 2.0 protocol, modified using human physiological data. This includes enzyme concentration calibration (e.g., pepsin, amylase, lipase), bile salt optimization, and kinetic digestion time points for whole food matrices. Human intubation studies will guide parameters to enhance physiological relevance. Proteolytic activity and digesta composition will be monitored by protein liquid chromatography (PLC), LC-MS, and time-resolved metabolomics profiling. Digested materials will be prepared under sterile conditions and applied to tissue constructs to assess tissue compatibility and stability.Aim 2: Intestinal Response and Functional EvaluationA bioengineered 3D human intestinal tissue model will be exposed to digested novel and control foods. This model includes organoid-derived epithelium, myofibroblasts, immune cells, neurons, and microbiota integrated into silk protein-based scaffolds. Exposure will be performed over short-term (Days 1, 3) and long-term (Weeks 1, 2) intervals. Functional readouts include:Cellular function and viability: Live/Dead, and Alamar Blue assays.Barrier integrity: FITC-dextran permeability, tight junction protein immunostaining.Inflammation and allergic responses: Multiplex cytokine arrays, ELISA, and immunophenotyping.Microbiome impact: 16S rRNA sequencing and SCFA profiling.Aim 3: Epigenetic Mechanism ProfilingOrganoid-derived epithelial cells will be repeatedly exposed to digested food products to simulate chronic dietary consumption. Key epigenetic changes will be monitored using:Chromatin accessibility: ATAC-seq.DNA methylation: Bisulfite sequencing.Histone modifications: ChIP-seq for H3K4me3, H3K27ac, and other relevant marks.Transcriptomic changes: RT-qPCR for specific genes.These data will be correlated with functional outcomes from Aim 2. Integration will reveal whether food-induced epigenetic changes correspond to altered inflammation, barrier integrity, or immune activation.Efforts to Cause a Change in Knowledge, Actions, or ConditionsResearch Mentorship: Undergraduate and graduate students will participate in laboratory rotations, thesis projects, and interdisciplinary training, gaining hands-on skills in tissue engineering, molecular biology, and food science.Curriculum Development: Project outputs will inform new modules in courses related to food safety, human physiology, microbiome health, and bioengineering.Publications and Presentations: Research findings will be shared through peer-reviewed journal articles, presentations at internal meetings and invited seminars, and participation in the NIFA Annual Grantees' Meeting.Stakeholder Engagement: Collaborations with the Friedman School of Nutrition Science and Policy at Tufts University, regulatory agencies (e.g., FDA) and food industry partners will ensure outputs align with translational needs.Open Access Protocols and Data: Protocols and data will be shared via public repositories and lab websites to encourage replication and adoption.Evaluation and Impact Assessment PlanTo assess the effectiveness and scientific impact of the project, both formative and summative evaluations will be conducted, including:Quantitative Scientific Output MetricsNumber of food types tested, models validated, and datasets generated.Publications in high-impact journals and citations.Research protocols uploaded to open-access databases (e.g., protocols.io).Number of invited talks, symposia contributions, and stakeholder consultations.Educational MetricsNumber of students trained, with tracking of outcomes such as thesis completions, conference presentations, and subsequent career placements.Student knowledge gain will be measured via pre- and post-training assessments in lab courses and workshops.Stakeholder and Industry EngagementDevelopment of partnership agreements for model testing or data use.Invitations for regulatory consultation.