The human gastrointestinal tract exhibits diverse and essential roles in processes as widespread as digestion, absorption, secretion and immunity. However, these same crucial functions represent a significant vulnerability by which bacterial and viral gastrointestinal pathogens induce substantial morbidity and mortality in humans worldwide. While animal models are invaluable to the study of basic mechanisms of infectious disease, the interactions between some of the most prominent gastrointestinal pathogens with host tissues are species specific and host restricted, particularly for infections that have co-evolved with humans. Thus, the study of many pathogens has heretofore required animal-adapted strains that do not accurately recapitulate the human disease pathophysiology and host range restriction in mice, or alternatively oncogenically transformed human cell lines that do not model normal tissue characteristics. Recent advances in culture of primary tissues as 3-dimensional organoids that reproduce multilineage differentiation and organ architecture represent a promising technology to modeling the interaction of human enteric pathogens with human gastrointestinal epithelium in vitro. The overall goal of this application is to create alternative model systems for a variety of human enteric diseases and pathogen-associated specific immune responses using optimized 3D human gastrointestinal organoid cultures via the co-culture of (1) primary gastric and intestinal organoids with (2) bacterial/viral pathogens and/or (3) immune effector cells. We have thus assembled a multidisciplinary, collaborative, and synergistic team at Stanford University including Calvin Kuo (gastrointestinal organoid culture), Manuel Amieva (Helicobacter pylori), Harry Greenberg (Rotavirus), Sarah Heilshorn (microenvironment bioengineering), Sean Bendall (CyTOF), Elizabeth Mellins (immunology), and Denise Monack (Salmonella Typhi). Together, these PIs form the Stanford Novel, Alternative Models for Enteric Diseases Cooperative Research Center (Stanford NAMSED CRC) Program. The Stanford NAMSED CRC is composed of Cores A-C (Administration, Organoid Production, and Advanced Co-Culture Engineering and Single Cell Statistics of Gut Immunology ACCESS-GI), and Projects 1-2, which study bacterial and viral pathogen interactions with human GI organoids, respectively. Towards these goals, we employ human organoid technologies incorporating epithelial-only or epithelial/mesenchymal components to create tissue-specific models of bacterial (Helicobacter pylori, Salmonella Typhi, Salmonella Typhimurium) or viral (rotavirus) infections, with or without immune effector cells or stimuli. We deploy innovative supporting technologies including CyTOF mass cytometry, bioengineered extracellular matrices, microfluidics and CRISPR-based gene editing. Overall, the Stanford NAMSED represents a synergistic group devoted to the integrated modeling of enteric pathogens as an epithelial-immune organoid unit.

Public Health Relevance

Bacterial and viral infections of the gastrointestinal tract are a leading cause of illness worldwide. Here, we grow human intestine and stomach as 3-dimensional 'organoids' and add gastrointestinal bacteria and viruses to create new disease models that could lead to improved treatments for these diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program--Cooperative Agreements (U19)
Project #
5U19AI116484-03
Application #
9221980
Study Section
Special Emphasis Panel (ZAI1-LG-M (J1))
Program Officer
Alarcon, Rodolfo M
Project Start
2015-03-01
Project End
2020-02-28
Budget Start
2017-03-01
Budget End
2018-02-28
Support Year
3
Fiscal Year
2017
Total Cost
$1,006,235
Indirect Cost
$308,078
Name
Stanford University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94304
Wosen, Jonathan E; Mukhopadhyay, Dhriti; Macaubas, Claudia et al. (2018) Epithelial MHC Class II Expression and Its Role in Antigen Presentation in the Gastrointestinal and Respiratory Tracts. Front Immunol 9:2144
Hartmann, Felix J; Simonds, Erin F; Bendall, Sean C (2018) A Universal Live Cell Barcoding-Platform for Multiplexed Human Single Cell Analysis. Sci Rep 8:10770
Ding, Siyuan; Zhu, Shu; Ren, Lili et al. (2018) Rotavirus VP3 targets MAVS for degradation to inhibit type III interferon expression in intestinal epithelial cells. Elife 7:
Haugh, Matthew G; Vaughan, Ted J; Madl, Christopher M et al. (2018) Investigating the interplay between substrate stiffness and ligand chemistry in directing mesenchymal stem cell differentiation within 3D macro-porous substrates. Biomaterials 171:23-33
Keren, Leeat; Bosse, Marc; Marquez, Diana et al. (2018) A Structured Tumor-Immune Microenvironment in Triple Negative Breast Cancer Revealed by Multiplexed Ion Beam Imaging. Cell 174:1373-1387.e19
LeSavage, Bauer L; Suhar, Nicholas A; Madl, Christopher M et al. (2018) Production of Elastin-like Protein Hydrogels for Encapsulation and Immunostaining of Cells in 3D. J Vis Exp :
Madl, Christopher M; Heilshorn, Sarah C; Blau, Helen M (2018) Bioengineering strategies to accelerate stem cell therapeutics. Nature 557:335-342
Nair, Nitya; Feng, Ningguo; Blum, Lisa K et al. (2017) VP4- and VP7-specific antibodies mediate heterotypic immunity to rotavirus in humans. Sci Transl Med 9:
Dubbin, Karen; Tabet, Anthony; Heilshorn, Sarah C (2017) Quantitative criteria to benchmark new and existing bio-inks for cell compatibility. Biofabrication 9:044102
Janda, Claudia Y; Dang, Luke T; You, Changjiang et al. (2017) Surrogate Wnt agonists that phenocopy canonical Wnt and ?-catenin signalling. Nature 545:234-237

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