A broad range of inflammatory gastrointestinal conditions impact millions of people in the United States and abroad. The etiology of the disease is multifactorial and heterogeneous, but the common underlying feature is an overactive immune system. Genetics, environmental factors, age, sex, race and even the composition of bacteria that reside in the gut lumen are associated with causation of gut inflammation. While some therapeutic advances have been made, a large number of patients do not respond to existing pharmaceutical interventions leaving many to have life-long morbidity. This provides strong rationale to improve existing therapies and discover alternative and novel approaches to deal with inflammatory conditions of the gut. Preclinical models used for drug research and discovery have been historically poor. They are comprised cancer cells that have low physiological relevance and do not possess the immune cell compartment, which is a missing pivotal component to achieve an accurate model of human inflammatory conditions. For these reasons, there is a need in the therapeutics marketplace for an in vitro intestinal platform that accurately recapitulates luminal-epithelial- immune cell axis of the intestines. To meet this need, Altis Biosystems Inc., an early-stage biotechnology company, has collaborated with scientists at academic laboratories to develop a novel, primary-stem cells-based, in vitro intestinal model (termed RepliGut). We have finished an SBIR Phase I program focused on transgenesis and gene editing of intestinal stem cells (ISCs) cultured on RepliGut. We developed reporter-gene ISCs that generate differentiated epithelium and sense and report in real-time key features of gut inflammation, 1) barrier function, and 2) NF-kB activation. Here we expand the scope to generate a first-in-kind co-culture model called, InflammaGutTM. InflammaGut is a tripartite and flexible system that uses fundamental technology of RepliGut and superimposes inflammation reporter genes and co-culture of human epithelium with allogeneic gut-associated lymphocytes (GAL) to recreate the LEI-axis. Four products are envisioned: 1) InflammaGut:ZO1 and InflammaGut:NF-kB reporter gene epithelium, 2) InflammaGut:Co-Culture: an epithelial/gut-associate lymphocyte (GAL) co-culture system, 3) commercializable human gut-associated lymphocytes, and 4) a new contract research service (CRS) using InflammaGut. There is strong engineering and biological method innovation, including, 1) development of Transwell insert enabling scalable culture of RepliGut differentiated human epithelium over a hydrogel encapsulated lymphoid-cell compartment, and 2) isolation, culture, and cryobanking of GAL from human organ donors. InflammaGut will be built, validated, and tested by academic collaborators (founders of the RepliGut technology), Altis Biosystems, and two industry collaborators. 1
Public Health Relevance Statement: Narrative: InflammaGutTM is a novel co-culture platform that mimics the Lumina-Epithelial-Lymphocyte axis using human stem cell-derived epithelium and gut-associated lymphocytes to accurately model gut inflammation and revolutionize pre-clinical culture systems for the pharmaceutical industry. 1
Project Terms: Affect; Age; ages; Bacteria; Biotechnology; Biotech; Cell Communication; Cell Interaction; Cell-to-Cell Interaction; Cell Compartmentation; Cell Compartmentations; Cell Differentiation process; Cell Differentiation; Cell Line; CellLine; Strains Cell Lines; cultured cell line; cell motility; Cell Locomotion; Cell Migration; Cell Movement; Cellular Migration; Cellular Motility; Motility; Cell Survival; Cell Viability; Cells; Cell Body; Client; Collagen; Colon; Colonic Neoplasms; Colon Neoplasms; Colon Tumor; Colonic Mass; Colonic Tumor; colon neoplasia; Cryopreservation; Cryofixation; cold preservation; cold storage; Disease; Disorder; Drug Design; Drug Industry; Pharmaceutic Industry; Pharmaceutical Industry; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Engineering; Environment; Epithelial Cells; Gastrointestinal Diseases; gastrointestinal disorder; Genes; Goals; Health; Human; Modern Man; Immune system; allergic/immunologic body system; allergic/immunologic organ system; In Vitro; Inflammation; Inflammatory Bowel Diseases; Inflammatory Bowel Disorder; inflammatory disease of the intestine; inflammatory disorder of the intestine; intestinal autoinflammation; Small Intestines; small bowel; Intestines; Intestinal; bowel; Laboratories; Lymphocyte; Lymphatic cell; Lymphocytic; lymph cell; Methods; Biological Models; Biologic Models; Model System; Morbidity - disease rate; Morbidity; Mucous Membrane; Mucosa; Mucosal Tissue; Persons; Organ Donor; Patients; Production; Proteins; QOL; Quality of life; Racial Group; Racial Stocks; Race; Research; Science; Cell Communication and Signaling; Cell Signaling; Intracellular Communication and Signaling; Signal Transduction Systems; Signaling; biological signal transduction; Signal Transduction; Societies; Progenitor Cells; stem cells; Technology; Testing; Time; tissue culture; Transplantation; transplant; United States; cytokine; NF-kappa B; Immunoglobulin Enhancer-Binding Protein; NF-kB; NF-kappaB; NFKB; Nuclear Factor kappa B; Nuclear Transcription Factor NF-kB; Transcription Factor NF-kB; kappa B Enhancer Binding Protein; nuclear factor kappa beta; Tumor Cell Line; Lymphoid Cell; Mediating; base; culture plates; improved; Lateral; Apical; Acute; Chronic; Encapsulated; Phase; Biological; biologic; Physiological; Physiologic; Reporter Genes; Evaluation; Epithelial; Individual; Withdrawal; Occluding Junctions; Zonula Occludens; Tight Junctions; drug use; Drug usage; inflammatory mediator; Inflammation Mediators; Co-culture; Cocultivation; Coculture; Coculture Techniques; Therapeutic; Genetic; Inflammatory; Malignant Cell; cancer cell; Reporter; Life; programs; Scientist; Immunes; Immune; Complex; Autologous; Event; Side; System; gastrointestinal; Services; American; cohort; Peripheral Blood Lymphocyte; Hydrogels; transgenic; Transgenic Organisms; Immunomodulation; immune modulation; immune regulation; immunologic reactivity control; immunomodulatory; immunoregulatory; immunoregulation; novel; drug market; General Public; General Population; Environmental Factor; environmental risk; Environmental Risk Factor; Reporting; Gut Epithelium; gastrointestinal epithelium; Modeling; High Throughput Assay; high throughput screening; monolayer; Intervention Strategies; interventional strategy; Intervention; Transgenesis; Gene Transfer Techniques; Pharmaceutical Agent; Pharmaceuticals; Pharmacological Substance; Pharmacologic Substance; Causality; causation; disease causation; Etiology; Symptoms; Detection; Dietary Factors; immune competent; Immunocompetent; Industry Collaborators; Industry Collaboration; Preclinical Models; Pre-Clinical Model; Reproducibility; Research Contracts; Allogenic; Small Business Innovation Research Grant; SBIR; Small Business Innovation Research; Validation; sex; Modification; Development; developmental; pre-clinical; preclinical; human stem cells; novel strategies; new approaches; novel approaches; novel strategy; innovation; innovate; innovative; novel therapeutics; new drug treatments; new drugs; new therapeutics; new therapy; next generation therapeutics; novel drug treatments; novel drugs; novel therapy; self-renewal; self-renew; demographics; commercialization; drug candidate; flexibility; flexible; phase 1 study; Phase I Study; screening; microbiota; microbial consortia; microbial flora; microflora; multispecies consortia; Drug Screening; human model; model of human; Financial Hardship; financial burden; financial distress; financial strain; financial stress; tumor-immune system interactions; immune microenvironment; immunosuppressive microenvironment; immunosuppressive tumor microenvironment; tumor immune microenvironment; gut inflammation; bowel inflammation; inflamed bowel; inflamed gut; inflamed intestine; intestinal inflammation
